DEVELOPMENT OF FUNCTIONAL BEVERAGE FROM BARLEY

By

AHMAD DIN MSc (Hons) Food Technology

Thesis submitted in partial fulfillment of requirements for the degree of

DOCTOR OF PHILOSOPHY IN

FOOD TECHNOLOGY

NATIONAL INSTITUTE OF FOOD SCIENCE AND TECHNOLOGY UNIVERSITY OF AGRICULTURE FAISALABAD

PAKISTAN

2009

Dedicated

to my Beloved Parents

ACKNOWLEDGMENTS

All praises and thanks are for Almighty Allah the Merciful the only creator of

the universe and source of all knowledge and wisdom who blessed me with health

thoughts talented teachers helping friends and opportunity to complete this study I

offer my humblest thanks to Holy Prophet Hazrat Muhammad (Peace be Upon Him)

whose moral and spiritual teachings enlightened my heart mind and flourished my

thoughts towards achieving high ideals of life

I am grateful to my supervisor Professor Dr Faqir Muhammad Anjum Director

General National Institute of Food Science and Technology University of Agriculture

Faisalabad for his supervision in planning execution and scholarly ideas that beautified

the scientific nature of the research work presented in this manuscript He always

directed to enlighten the ways of life as well

I am thankful to the members of my supervisory committee Dr Tahir Zahoor

Associate Professor National Institute of Food Science and Technology and Dr Haq

Nawaz Institute of Animal Nutrition and Feed Technology for their kind help and

constructive criticism during the course of study for the accomplishment of this piece of

work

I am also very grateful to all my teachers of the National Institute of Food Science

and Technology University of Agriculture Faisalabad for their technical guidance

moral support and prayers to accomplish this study

My sincere gratitude is to all my friends especially Moazzam Rafiq Khan Dr

Muhammad Saeed Dr Shahzad Hussain Dr Umair Arshad and rest of the Ph D club

and juniors especially Muhammad Nadeem who always helped me to complete my

work I feel it incomplete if I do not extend my fervent thanks and heartiest compliments

to my father and mother aunties brothers and sisters bhabis cousins nephews and

nieces for remembering me in their prayers and whose act always enforced me to update

my knowledge

(AHMAD DIN)

CC OO NN TT EE NN TT SS

CHAPTER TITLE PAGE

ABSTRACT

1 INTRODUCTION 1

2 REVIEW OF LITERATURE 6

3 MATERIALS AND METHODS 39

4 RESULTS AND DISCUSSION 56

5 SUMMARY 131

CONCLUSIONS 136

RECOMMENDATIONS 137

LITERATURE CITED 138

APPENDICES 162

List of Tables

Table Title Page

31 Treatment plan 46 32 Different treatments used in the biological study 54 41 Chemical composition of barley flour 57 42 Chemical Analysis of β-glucan 59 43 Mean sum of squares for color values (L a b) of stored β-glucan

beverages 62

44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

63

45 Effect of treatments and storage intervals on the a-value of stored β-glucan beverages

66

46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

68

47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

71

48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

72

49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

75

410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

76

411 Mean sum of squares for pH acidity and ascorbic acid content of stored beverages

78

412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

79

413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

82

414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

84

415 Mean sum of squares for reducing non reducing and total sugar content of stored beverages

87

416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

88

417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

90

418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

92

Table Title Page

419 Effect of treatments and storage intervals on the total plate count

(CFUml) of stored β-glucan beverages 94

420 Mean sum of squares for sensory evaluation of stored beverages 96 421 Effect of treatments and storage intervals on the color score of

stored β-glucan beverages 97

422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

100

423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

103

424 Effect of treatments and storage intervals and on the sourness score of stored β-glucan beverages

105

425 Effect of storage intervals and treatments on the overall acceptability score of stored β-glucan beverages

108

426 Mean sum of squares for blood lipid profile of volunteers 110 427 Effect of β-glucan supplemented beverage on serum total

cholesterol content (mgdl) of healthy subjects 111

428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

115

429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

119

430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

123

431 Mean sum of squares for blood glucose contents of volunteers 127 432 Effect of β-glucan beverage on blood glucose (mgdl) content of

with different time intervals 127

433 Interactive effect of diets and time scale intervals on the blood glucose contents (mgdl) of volunteers

127

434 Interactive effect of diets and study duration on the blood glucose contents (mgdl) of volunteers

128

List of Figures

Fig Title Page

31 Preparation of β -glucan beverage 47 41 Percent decrease in the serum total cholesterol level of subjects fed

on different beverages 111

42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of healthy volunteers

112

43 Percent decrease in the serum triglycerides level of subjects fed on different beverages

115

44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy volunteers

116

45 Percent decrease in the serum LDL level of subjects fed on different beverages

119

46 Effect of β-glucan beverage on LDL (mgdl) content of healthy volunteers

120

47 Percent increase in the serum HDL level of subjects fed on different beverages

123

48 Effect of β-glucan beverage on HDL (mgdl) content of healthy volunteers

124

49 Effect of β-glucan beverage on blood glucose (mgdl) content of healthy volunteers

128

List of Appendices

Appendix Title Page

I Composit ion of functional beverage 162

II 9 Point Hedonic Scale 163

III Food frequency questionnaire 164

IV Demographic information performa (subjects) 165

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

LITERATURE CITED

AACC 2000 Approved Methods of American Association of Cereal Chemists The American Association of Cereal Chemists Inc St Paul Minnesota USA

Aastrup S 1979a The effect of rain on β -glucan content in barley grains Carlsberg esearch Communications 44381-393

Aditya K T Yokota S Suzuki and H Etoh 2008 Sub crit ical Water Extraction of Barley to Produce a Functional Drink

Biosci Biotechnol Biochem 72(1)236-239

AERI 1896 The Agricultural Economics Research Institute Balance Sheet for Food Commodities Finland 1985 The Insti tute Helsinki

Akubor PI 2003 Influence of storage on the physicochemical microbiological and sensory properties of heat and chemically treated melon-banana beverage Plant Foods for Human Nutri 58 1ndash10

Alessandra DC P Antonio V Vincenzo A Mario 2004 Changes of f lavonoids vitamin C and antioxidant capacity in minimally processed citrus segments and juices during storage Food Chem 84 99-105

Aman P H Graham AC Til ly 1989 Content and solubil i ty of mixed-l inked (1-3) (1-4)- β -D-glucan in barley and oats during kernel development and storage J Cereal Sci 1045-50

Anderson J W 1980 Dietary f iber and diabetes in Medical Aspects of Dietry Fiber G A spil ler and R M Key eds Plenum Medical Book Company New York

Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

Anderson J W DB Spencer CC Hamilton SF Smith and J Tietyen CA Bryant P Oeltgen 1990 Oat-bran cereal lowers serum total and LDL cholesterol in hypercholesterolemic men Am J Clin Nutri 52 495-499

139

Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

Annoni G BM Botasso D Ciaci MF Donato and A Tripodi 1982 Liquid tr iglycerides (GPO-PAP) Medi Diagnostic I taly Lab J Res Lab Med 9 115-116

AOAC 2000 Official Methods of Analysis The Association of the Official Analytical Chemists 20 t h Ed Arlington USA

Arndt EA 2006 Whole-grain barley for todays health and wellness needs ConAgra Foods Inc Omaha NE 51(1) 20-22

Assmann G 1979 HDL-cholesterol precipitant Randox Labs Ltd CrumLin Co Antrim N Ireland Internist 20559-567

Babsky NE J L Toribio and J E Lozano 1986 Influence of storage on the composit ion of clarif ied apple juice concentrate J Food Sci 51 (3) 564-67

Ballance GM WOS Meredith 1976 Purif ication and partial characterization of an endo- β -13-glucanase from green malt J Inst Brew 8264-67

Bamforth CW and AHP Barclay 1993 Malting technology and the uses of malt In Barley Chemistry and Technology (eds AW MacGregor and RS Bhatty) by Am Assoc Cereal Chem St Paul USA pp 297-354

Bansema C 2000 Development of a barley P-glucan beverage with and without whey protein Isolate MSc thesis Edmonton Alberta Canada

Basman A and HK Ksel 1999 Properties and composit ion of Turkish f lat bread (bazlama) supplemented with barley f lour and wheat bran Cereal Chem 76506ndash511

Beer MU E Arrigoni and R Amado 1995 Effect of oat gum on blood cholesterol levels in healthy young men Europ J Clin Nutri 49517ndash522

140

Beer MU PJ Wood J Weisz N Fi l l ion 1997 Effect of cooking and storage on the amount and molecular weight of (1rarr3) (1rarr4) - β -D-glucan extracted from oat products by an in vitro digestion system Cereal Chem 74 705-709

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 A Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bender DA and AE Bender 1999 Bendersrsquo Dictionary of Nutrit ion and Food Technology 7 t h ed Woodhead Publishing Abington

Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

Berglund PT CE Fastnaught ET Holm 1992 Food uses of waxy hull- less barley Cereal Foods World 37707ndash714

Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

Bhatty RS 1992 Total and extractable β -glucan contents of oats and their relationship to viscosity J Cer Sci 15185-192

Bhatty RS 1995 Laboratory and pilot plant extraction and purif ication of b-glucans from hull- less barley and oat bran J Cer Sci 22163ndash170

Bhatty RS 1996 Production of food malt from hull- less barley Cereal Chem 73(1) 75-80

Bhatty RS AW MacGregor and BG Rossnagel 1991 Total and acid-soluble β -glucan content of hulless barley and its relationship to acid-extract viscosity Cereal Chem 68221-227

Bhatty RS1986 Physiochemical and Functional (Breadmaking) Properties of Hull- less Barley Fractions Cereal Chem 6331-35

141

Bibek R 2001 Fundamental Food Microbiology 2nd edn The CRC press Ltd Washington DC pp 56-90

Bingham SA NE Day R Luben P Ferrari N Sl imani T Norat F Lavel E Kesse A Nieters H Boeing A Tjoslashnneland K Overvad C Martinez M Dorrensoro CA Gonzalez TJ Key A Trichopoulou A Naska P Vineis R Tumino V Krogh HB Bueno-de-Mesquita PHM Peeters G Berglung G Hallmans E Lund G Skele R Kaaks and E Riboli 2003 Dietary f ibre in food and protection against colorectal cancer in the European Prospective Investigation into Cancer and Nutrit ion (EPIC) an observational study Lancet 3611496-501

Bioumlrklund M A van Rees RP Mensink and G Oumlnning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucans from oats and barley a randomised dose-controlled tr ial Eur J Clin Nutri 591272-1281

Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

Bjorck I AC Eliasson A Drews M Gudmundsson and R Karlsson 1990 some nutrit ional properties of starch and dietary f iber in barley genotypes containing different levels of amylose Cereal Chem 67 327

BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

Brand J S Colagiuri S Crossman A Allen D Roberts and S Truswell 1991 Low-glycemic index foods improve long term glycemic control in NIDDM Diabetes Care 14 95ndash101

142

Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

Brennan CS and LJ Cleary 2005 The potential use of cereal (1314)- β -D-glucans as functional food ingredients J Cer Sci 421ndash13

Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

Brown L B Rosner W Willet and FM Sacks 1999 Cholesterol lowering effects of dietary f iber a meta analysis Am J Clin Nutri 69 (1) 30 42

Brunswick P DJ Manner and J K Stark 1987 Development of β -D-glucanases during germination of barley and the effect of ki lning on individual isoenzymes J Inst Brew 93181-186

Bryan D J Robert AT Wilson T Carlson S Frazer GH Zheng 2003 β -Glucan Fractions from Barley and Oats Are Similarly Antiatherogenic in Hypercholesterolemic Syrian Golden Hamsters The American Society for Nutrit ional Sciences J Nutri Metabolism 133468-475

Buliga GS DA Brant and GB Fincher 1986 The sequence statist ics and solution configration of barley (1rarr3) (1rarr4) - β -D-glucan Carbohydr Res 57139-156

Burkus Z 1996 Barley P-Glucan Extraction Functional Properties and Interactions with Food Components MSc thesis Edmonton AlbertaCanda

Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloid F Glicksman M (Ed) p 49-93 CRC Press Inc Boca Raton

Burkus Z 1996 Barley β -glucan Extraction Functional properties and interaction with food components MSc Thesis Dept of Agricultural Food and Nutrit ional Science Univ of Alberta Edmonton Canada

143

Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

Burkus Z and F Temell i 1999 Glucan concentrate J Food Sci 64198-201 Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloidr Glicksman M (Ed) p 49-93 CRC Press hc Boca Raton FL

Burkus Z and F Temell i 2005 Rheological properties of barley β -glucan Carbohydr Polym 59 459ndash465

Burkus Z F Temell i 1999 Gelation of barley β -glucan - concentrate J Food Sci 64198-201

Calix FD and N Bardrie 2004 Consumer acceptance and physicochemical quality of processed red sorrelroselle (Hibiscus sabdar i f fa L) sauces from enzymatic extracted calyces 4 141-148

Carpita NC 1996 Structure and biogenesis of cel l walls of grasses Annual Rev Plant Physiol Plat Molecular Biol 47445-476

Carr J M S Glatter J L Jeraci and B A Lewis 1990 Enzymes Determination of Beta-Glucan in Cereal-Based Food Products Cereal Chem 67226-229

Casterl ine J L CJ Oles and Y Ku 1997 In vitro fermentation of various food f iber reactions J Agric Food Chem 452463ndash2467

Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

Chowdhury MGF MN Islam MS Is lam T Is lam and MS Hossain 2008 Study on Preparation and Shelf-Life of Mixed Juice Based on Wood Apple and Papaya J Soil Nature 2(3) 50-60

Chung OK and Y Pomeranz 1985 Amino acids in cereal proteins and protein fractions Ch 5 in Digesfibi l i~ and

144

Amino Acid Availabil i ty in Cereals andOilseeds J W Finley and DT Hopkins (Eds) pp 169-232 AACC St Paul MN

Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

Crandall PG CS Chen and KC Davis 1987 Preparation and storage of 72 brix orange juice concentration J Food Sci 52 (3) 381

Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

Daw ZY YSA El-Gizaw and AMB Said 1994 Microbiological evaluation of some local juices and drinks Chemie Mikrobiologie Technologie der Lebensmittel 168ndash15

Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

Dawkins NL I A Nnanna 1993 Studies on oat gum [(1rarr31rarr4)- β-D-glucan] Composit ion molecular weight est imation and rheological properties Food Hydrocol 9 1-7

Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

Dohnalek MH 2004 The role of f ibre in cl inical nutrit ion In Van der Kamp JW Asp NG Miller J J Schaafsma G (Ed) Dietary f ibre bioactive carbohydrates for food and feed Wageningen Academic Publishers Wageningen pp 271294

145

Dongowski G M Huth E Gebhardt and W Flamme 2002 Dietary f iber-rich barley products beneficial ly affect the intestinal tract of rats J Nutri 132(12) 3704-14

Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

Falade OS OR Sowunmi A Oladipo A Tobosun and SRA Adewusi 2003 The level of organic acids in some Nigerian fruit and their effect on mineral availabil i ty in composite diet Pak J Nutri 2(2) 82-83

Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

FDA 1996 Food labeling Health claims oats and coronary heart disease Federal Register 61 (3) January 4

Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

Frazier WC and EM Foster 1958 Laboratory Manual for Food Microbiology Burgess Pub Co Minneapolis Minnisota USA

Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

146

Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

Giese J H 1992 Hitt ing the spot Beverages and beverage technology Food Technol 4670-72 74-75 78-80

Godara RK and OP Pareek 1985 Effect of temperature in storage of ready to serve date juice beverages indian j agric Sci 55 (5) 347-349 (FSTA 18 (4) 78 1986)

Gonzalez ER and S Leeson 2000 An investigation on the preservation of kununndashzaki an African fermented cereal based food drink Acta Alimentaria 29 385ndash92

GOP 2008 Government of Pakistan Finance Division Economic Advisor s Wing Islamabad Pakistan

Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

Hallfr isch J DJ Schofield KM Behall 2003 Physiological responses of men and women to barley and oat extracts (NutrimX) I I Comparison of glucose and insulin responses Cereal Chem 8080ndash83

Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

147

Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

Lawless HT and H heymann Sensory evaluation of food Principles and Practices Gaithersburg MD Aspen Publishers ISSN 1572-0330) Oorspr uitg New York [etc ] Chapman amp Hall 1998

Hashimoto S MD Shogren Y Pomeranz 1987 Cereal Pentosans Their est imation and signif icance I Pentosans in wheat and milled wheat products Cereal Chem 64(1) 30-34

Hassan SA 1976 Effect of storage on physico-chemical characterist ics of carbonated orange juice Msc thesis Food Tech Deptt WPAU Lyallpur

Hatcher WSJ R J L Weihe DF Split tstoesser EC Hil l and ME Parish 1992 Fruit Beverages In Compendium of methods for the microbiological examination of foods Vanderzant C Split tstoesser DF (eds) American Public Health Association Washington DC

Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

Hil l M J and FR Path 1998 Cereals dietary f iber and cancer Nutri Res 18563ndash659

Hil l iam M 2000 Functional foodndashndashHow big is the market The World of Food Ingredients 12 50ndash2

Holsinger V H LP Posati and ED DeVilbiss 1974 Whey beverages a review J Dairy Sci 57(7) 849ndash859

Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

148

47ndash82) St Paul MN USA American Association of Cereal Chemists

Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

Jadhav SJ S E Lutz VM Ghorpade and DK Salunkhe 1998 Barley chemistry and value-added processing Crit ical Rev Food Sci 3823ndash171

Jal i l i T REC Wildman DM Medeiros 2000 Nutraceutical roles of dietary f iber J Nutraceutical functional and Medi foods 2 19-34

Jansen MC HB Bueno-de-Mesquita R Buzina F Fidanza A Menotti H Blackburn AM Nissinen FJ Kok D Kromhout 1999 Dietary f iber and plant foods in relation to colorectal cancer mortal i ty The Seven Countries Study Inter J Canc 81 174-179

Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high

Dedicated

to my Beloved Parents

ACKNOWLEDGMENTS

All praises and thanks are for Almighty Allah the Merciful the only creator of

the universe and source of all knowledge and wisdom who blessed me with health

thoughts talented teachers helping friends and opportunity to complete this study I

offer my humblest thanks to Holy Prophet Hazrat Muhammad (Peace be Upon Him)

whose moral and spiritual teachings enlightened my heart mind and flourished my

thoughts towards achieving high ideals of life

I am grateful to my supervisor Professor Dr Faqir Muhammad Anjum Director

General National Institute of Food Science and Technology University of Agriculture

Faisalabad for his supervision in planning execution and scholarly ideas that beautified

the scientific nature of the research work presented in this manuscript He always

directed to enlighten the ways of life as well

I am thankful to the members of my supervisory committee Dr Tahir Zahoor

Associate Professor National Institute of Food Science and Technology and Dr Haq

Nawaz Institute of Animal Nutrition and Feed Technology for their kind help and

constructive criticism during the course of study for the accomplishment of this piece of

work

I am also very grateful to all my teachers of the National Institute of Food Science

and Technology University of Agriculture Faisalabad for their technical guidance

moral support and prayers to accomplish this study

My sincere gratitude is to all my friends especially Moazzam Rafiq Khan Dr

Muhammad Saeed Dr Shahzad Hussain Dr Umair Arshad and rest of the Ph D club

and juniors especially Muhammad Nadeem who always helped me to complete my

work I feel it incomplete if I do not extend my fervent thanks and heartiest compliments

to my father and mother aunties brothers and sisters bhabis cousins nephews and

nieces for remembering me in their prayers and whose act always enforced me to update

my knowledge

(AHMAD DIN)

CC OO NN TT EE NN TT SS

CHAPTER TITLE PAGE

ABSTRACT

1 INTRODUCTION 1

2 REVIEW OF LITERATURE 6

3 MATERIALS AND METHODS 39

4 RESULTS AND DISCUSSION 56

5 SUMMARY 131

CONCLUSIONS 136

RECOMMENDATIONS 137

LITERATURE CITED 138

APPENDICES 162

List of Tables

Table Title Page

31 Treatment plan 46 32 Different treatments used in the biological study 54 41 Chemical composition of barley flour 57 42 Chemical Analysis of β-glucan 59 43 Mean sum of squares for color values (L a b) of stored β-glucan

beverages 62

44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

63

45 Effect of treatments and storage intervals on the a-value of stored β-glucan beverages

66

46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

68

47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

71

48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

72

49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

75

410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

76

411 Mean sum of squares for pH acidity and ascorbic acid content of stored beverages

78

412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

79

413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

82

414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

84

415 Mean sum of squares for reducing non reducing and total sugar content of stored beverages

87

416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

88

417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

90

418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

92

Table Title Page

419 Effect of treatments and storage intervals on the total plate count

(CFUml) of stored β-glucan beverages 94

420 Mean sum of squares for sensory evaluation of stored beverages 96 421 Effect of treatments and storage intervals on the color score of

stored β-glucan beverages 97

422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

100

423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

103

424 Effect of treatments and storage intervals and on the sourness score of stored β-glucan beverages

105

425 Effect of storage intervals and treatments on the overall acceptability score of stored β-glucan beverages

108

426 Mean sum of squares for blood lipid profile of volunteers 110 427 Effect of β-glucan supplemented beverage on serum total

cholesterol content (mgdl) of healthy subjects 111

428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

115

429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

119

430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

123

431 Mean sum of squares for blood glucose contents of volunteers 127 432 Effect of β-glucan beverage on blood glucose (mgdl) content of

with different time intervals 127

433 Interactive effect of diets and time scale intervals on the blood glucose contents (mgdl) of volunteers

127

434 Interactive effect of diets and study duration on the blood glucose contents (mgdl) of volunteers

128

List of Figures

Fig Title Page

31 Preparation of β -glucan beverage 47 41 Percent decrease in the serum total cholesterol level of subjects fed

on different beverages 111

42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of healthy volunteers

112

43 Percent decrease in the serum triglycerides level of subjects fed on different beverages

115

44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy volunteers

116

45 Percent decrease in the serum LDL level of subjects fed on different beverages

119

46 Effect of β-glucan beverage on LDL (mgdl) content of healthy volunteers

120

47 Percent increase in the serum HDL level of subjects fed on different beverages

123

48 Effect of β-glucan beverage on HDL (mgdl) content of healthy volunteers

124

49 Effect of β-glucan beverage on blood glucose (mgdl) content of healthy volunteers

128

List of Appendices

Appendix Title Page

I Composit ion of functional beverage 162

II 9 Point Hedonic Scale 163

III Food frequency questionnaire 164

IV Demographic information performa (subjects) 165

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

LITERATURE CITED

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Aastrup S 1979a The effect of rain on β -glucan content in barley grains Carlsberg esearch Communications 44381-393

Aditya K T Yokota S Suzuki and H Etoh 2008 Sub crit ical Water Extraction of Barley to Produce a Functional Drink

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AERI 1896 The Agricultural Economics Research Institute Balance Sheet for Food Commodities Finland 1985 The Insti tute Helsinki

Akubor PI 2003 Influence of storage on the physicochemical microbiological and sensory properties of heat and chemically treated melon-banana beverage Plant Foods for Human Nutri 58 1ndash10

Alessandra DC P Antonio V Vincenzo A Mario 2004 Changes of f lavonoids vitamin C and antioxidant capacity in minimally processed citrus segments and juices during storage Food Chem 84 99-105

Aman P H Graham AC Til ly 1989 Content and solubil i ty of mixed-l inked (1-3) (1-4)- β -D-glucan in barley and oats during kernel development and storage J Cereal Sci 1045-50

Anderson J W 1980 Dietary f iber and diabetes in Medical Aspects of Dietry Fiber G A spil ler and R M Key eds Plenum Medical Book Company New York

Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

Anderson J W DB Spencer CC Hamilton SF Smith and J Tietyen CA Bryant P Oeltgen 1990 Oat-bran cereal lowers serum total and LDL cholesterol in hypercholesterolemic men Am J Clin Nutri 52 495-499

139

Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

Annoni G BM Botasso D Ciaci MF Donato and A Tripodi 1982 Liquid tr iglycerides (GPO-PAP) Medi Diagnostic I taly Lab J Res Lab Med 9 115-116

AOAC 2000 Official Methods of Analysis The Association of the Official Analytical Chemists 20 t h Ed Arlington USA

Arndt EA 2006 Whole-grain barley for todays health and wellness needs ConAgra Foods Inc Omaha NE 51(1) 20-22

Assmann G 1979 HDL-cholesterol precipitant Randox Labs Ltd CrumLin Co Antrim N Ireland Internist 20559-567

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Ballance GM WOS Meredith 1976 Purif ication and partial characterization of an endo- β -13-glucanase from green malt J Inst Brew 8264-67

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Bansema C 2000 Development of a barley P-glucan beverage with and without whey protein Isolate MSc thesis Edmonton Alberta Canada

Basman A and HK Ksel 1999 Properties and composit ion of Turkish f lat bread (bazlama) supplemented with barley f lour and wheat bran Cereal Chem 76506ndash511

Beer MU E Arrigoni and R Amado 1995 Effect of oat gum on blood cholesterol levels in healthy young men Europ J Clin Nutri 49517ndash522

140

Beer MU PJ Wood J Weisz N Fi l l ion 1997 Effect of cooking and storage on the amount and molecular weight of (1rarr3) (1rarr4) - β -D-glucan extracted from oat products by an in vitro digestion system Cereal Chem 74 705-709

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 A Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bender DA and AE Bender 1999 Bendersrsquo Dictionary of Nutrit ion and Food Technology 7 t h ed Woodhead Publishing Abington

Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

Berglund PT CE Fastnaught ET Holm 1992 Food uses of waxy hull- less barley Cereal Foods World 37707ndash714

Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

Bhatty RS 1992 Total and extractable β -glucan contents of oats and their relationship to viscosity J Cer Sci 15185-192

Bhatty RS 1995 Laboratory and pilot plant extraction and purif ication of b-glucans from hull- less barley and oat bran J Cer Sci 22163ndash170

Bhatty RS 1996 Production of food malt from hull- less barley Cereal Chem 73(1) 75-80

Bhatty RS AW MacGregor and BG Rossnagel 1991 Total and acid-soluble β -glucan content of hulless barley and its relationship to acid-extract viscosity Cereal Chem 68221-227

Bhatty RS1986 Physiochemical and Functional (Breadmaking) Properties of Hull- less Barley Fractions Cereal Chem 6331-35

141

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Bingham SA NE Day R Luben P Ferrari N Sl imani T Norat F Lavel E Kesse A Nieters H Boeing A Tjoslashnneland K Overvad C Martinez M Dorrensoro CA Gonzalez TJ Key A Trichopoulou A Naska P Vineis R Tumino V Krogh HB Bueno-de-Mesquita PHM Peeters G Berglung G Hallmans E Lund G Skele R Kaaks and E Riboli 2003 Dietary f ibre in food and protection against colorectal cancer in the European Prospective Investigation into Cancer and Nutrit ion (EPIC) an observational study Lancet 3611496-501

Bioumlrklund M A van Rees RP Mensink and G Oumlnning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucans from oats and barley a randomised dose-controlled tr ial Eur J Clin Nutri 591272-1281

Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

Bjorck I AC Eliasson A Drews M Gudmundsson and R Karlsson 1990 some nutrit ional properties of starch and dietary f iber in barley genotypes containing different levels of amylose Cereal Chem 67 327

BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

Brand J S Colagiuri S Crossman A Allen D Roberts and S Truswell 1991 Low-glycemic index foods improve long term glycemic control in NIDDM Diabetes Care 14 95ndash101

142

Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

Brennan CS and LJ Cleary 2005 The potential use of cereal (1314)- β -D-glucans as functional food ingredients J Cer Sci 421ndash13

Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

Brown L B Rosner W Willet and FM Sacks 1999 Cholesterol lowering effects of dietary f iber a meta analysis Am J Clin Nutri 69 (1) 30 42

Brunswick P DJ Manner and J K Stark 1987 Development of β -D-glucanases during germination of barley and the effect of ki lning on individual isoenzymes J Inst Brew 93181-186

Bryan D J Robert AT Wilson T Carlson S Frazer GH Zheng 2003 β -Glucan Fractions from Barley and Oats Are Similarly Antiatherogenic in Hypercholesterolemic Syrian Golden Hamsters The American Society for Nutrit ional Sciences J Nutri Metabolism 133468-475

Buliga GS DA Brant and GB Fincher 1986 The sequence statist ics and solution configration of barley (1rarr3) (1rarr4) - β -D-glucan Carbohydr Res 57139-156

Burkus Z 1996 Barley P-Glucan Extraction Functional Properties and Interactions with Food Components MSc thesis Edmonton AlbertaCanda

Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloid F Glicksman M (Ed) p 49-93 CRC Press Inc Boca Raton

Burkus Z 1996 Barley β -glucan Extraction Functional properties and interaction with food components MSc Thesis Dept of Agricultural Food and Nutrit ional Science Univ of Alberta Edmonton Canada

143

Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

Burkus Z and F Temell i 1999 Glucan concentrate J Food Sci 64198-201 Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloidr Glicksman M (Ed) p 49-93 CRC Press hc Boca Raton FL

Burkus Z and F Temell i 2005 Rheological properties of barley β -glucan Carbohydr Polym 59 459ndash465

Burkus Z F Temell i 1999 Gelation of barley β -glucan - concentrate J Food Sci 64198-201

Calix FD and N Bardrie 2004 Consumer acceptance and physicochemical quality of processed red sorrelroselle (Hibiscus sabdar i f fa L) sauces from enzymatic extracted calyces 4 141-148

Carpita NC 1996 Structure and biogenesis of cel l walls of grasses Annual Rev Plant Physiol Plat Molecular Biol 47445-476

Carr J M S Glatter J L Jeraci and B A Lewis 1990 Enzymes Determination of Beta-Glucan in Cereal-Based Food Products Cereal Chem 67226-229

Casterl ine J L CJ Oles and Y Ku 1997 In vitro fermentation of various food f iber reactions J Agric Food Chem 452463ndash2467

Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

Chowdhury MGF MN Islam MS Is lam T Is lam and MS Hossain 2008 Study on Preparation and Shelf-Life of Mixed Juice Based on Wood Apple and Papaya J Soil Nature 2(3) 50-60

Chung OK and Y Pomeranz 1985 Amino acids in cereal proteins and protein fractions Ch 5 in Digesfibi l i~ and

144

Amino Acid Availabil i ty in Cereals andOilseeds J W Finley and DT Hopkins (Eds) pp 169-232 AACC St Paul MN

Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

Crandall PG CS Chen and KC Davis 1987 Preparation and storage of 72 brix orange juice concentration J Food Sci 52 (3) 381

Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

Daw ZY YSA El-Gizaw and AMB Said 1994 Microbiological evaluation of some local juices and drinks Chemie Mikrobiologie Technologie der Lebensmittel 168ndash15

Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

Dawkins NL I A Nnanna 1993 Studies on oat gum [(1rarr31rarr4)- β-D-glucan] Composit ion molecular weight est imation and rheological properties Food Hydrocol 9 1-7

Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

Dohnalek MH 2004 The role of f ibre in cl inical nutrit ion In Van der Kamp JW Asp NG Miller J J Schaafsma G (Ed) Dietary f ibre bioactive carbohydrates for food and feed Wageningen Academic Publishers Wageningen pp 271294

145

Dongowski G M Huth E Gebhardt and W Flamme 2002 Dietary f iber-rich barley products beneficial ly affect the intestinal tract of rats J Nutri 132(12) 3704-14

Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

Falade OS OR Sowunmi A Oladipo A Tobosun and SRA Adewusi 2003 The level of organic acids in some Nigerian fruit and their effect on mineral availabil i ty in composite diet Pak J Nutri 2(2) 82-83

Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

FDA 1996 Food labeling Health claims oats and coronary heart disease Federal Register 61 (3) January 4

Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

Frazier WC and EM Foster 1958 Laboratory Manual for Food Microbiology Burgess Pub Co Minneapolis Minnisota USA

Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

146

Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

Giese J H 1992 Hitt ing the spot Beverages and beverage technology Food Technol 4670-72 74-75 78-80

Godara RK and OP Pareek 1985 Effect of temperature in storage of ready to serve date juice beverages indian j agric Sci 55 (5) 347-349 (FSTA 18 (4) 78 1986)

Gonzalez ER and S Leeson 2000 An investigation on the preservation of kununndashzaki an African fermented cereal based food drink Acta Alimentaria 29 385ndash92

GOP 2008 Government of Pakistan Finance Division Economic Advisor s Wing Islamabad Pakistan

Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

Hallfr isch J DJ Schofield KM Behall 2003 Physiological responses of men and women to barley and oat extracts (NutrimX) I I Comparison of glucose and insulin responses Cereal Chem 8080ndash83

Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

147

Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

Lawless HT and H heymann Sensory evaluation of food Principles and Practices Gaithersburg MD Aspen Publishers ISSN 1572-0330) Oorspr uitg New York [etc ] Chapman amp Hall 1998

Hashimoto S MD Shogren Y Pomeranz 1987 Cereal Pentosans Their est imation and signif icance I Pentosans in wheat and milled wheat products Cereal Chem 64(1) 30-34

Hassan SA 1976 Effect of storage on physico-chemical characterist ics of carbonated orange juice Msc thesis Food Tech Deptt WPAU Lyallpur

Hatcher WSJ R J L Weihe DF Split tstoesser EC Hil l and ME Parish 1992 Fruit Beverages In Compendium of methods for the microbiological examination of foods Vanderzant C Split tstoesser DF (eds) American Public Health Association Washington DC

Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

Hil l M J and FR Path 1998 Cereals dietary f iber and cancer Nutri Res 18563ndash659

Hil l iam M 2000 Functional foodndashndashHow big is the market The World of Food Ingredients 12 50ndash2

Holsinger V H LP Posati and ED DeVilbiss 1974 Whey beverages a review J Dairy Sci 57(7) 849ndash859

Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

148

47ndash82) St Paul MN USA American Association of Cereal Chemists

Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

Jadhav SJ S E Lutz VM Ghorpade and DK Salunkhe 1998 Barley chemistry and value-added processing Crit ical Rev Food Sci 3823ndash171

Jal i l i T REC Wildman DM Medeiros 2000 Nutraceutical roles of dietary f iber J Nutraceutical functional and Medi foods 2 19-34

Jansen MC HB Bueno-de-Mesquita R Buzina F Fidanza A Menotti H Blackburn AM Nissinen FJ Kok D Kromhout 1999 Dietary f iber and plant foods in relation to colorectal cancer mortal i ty The Seven Countries Study Inter J Canc 81 174-179

Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high

ACKNOWLEDGMENTS

All praises and thanks are for Almighty Allah the Merciful the only creator of

the universe and source of all knowledge and wisdom who blessed me with health

thoughts talented teachers helping friends and opportunity to complete this study I

offer my humblest thanks to Holy Prophet Hazrat Muhammad (Peace be Upon Him)

whose moral and spiritual teachings enlightened my heart mind and flourished my

thoughts towards achieving high ideals of life

I am grateful to my supervisor Professor Dr Faqir Muhammad Anjum Director

General National Institute of Food Science and Technology University of Agriculture

Faisalabad for his supervision in planning execution and scholarly ideas that beautified

the scientific nature of the research work presented in this manuscript He always

directed to enlighten the ways of life as well

I am thankful to the members of my supervisory committee Dr Tahir Zahoor

Associate Professor National Institute of Food Science and Technology and Dr Haq

Nawaz Institute of Animal Nutrition and Feed Technology for their kind help and

constructive criticism during the course of study for the accomplishment of this piece of

work

I am also very grateful to all my teachers of the National Institute of Food Science

and Technology University of Agriculture Faisalabad for their technical guidance

moral support and prayers to accomplish this study

My sincere gratitude is to all my friends especially Moazzam Rafiq Khan Dr

Muhammad Saeed Dr Shahzad Hussain Dr Umair Arshad and rest of the Ph D club

and juniors especially Muhammad Nadeem who always helped me to complete my

work I feel it incomplete if I do not extend my fervent thanks and heartiest compliments

to my father and mother aunties brothers and sisters bhabis cousins nephews and

nieces for remembering me in their prayers and whose act always enforced me to update

my knowledge

(AHMAD DIN)

CC OO NN TT EE NN TT SS

CHAPTER TITLE PAGE

ABSTRACT

1 INTRODUCTION 1

2 REVIEW OF LITERATURE 6

3 MATERIALS AND METHODS 39

4 RESULTS AND DISCUSSION 56

5 SUMMARY 131

CONCLUSIONS 136

RECOMMENDATIONS 137

LITERATURE CITED 138

APPENDICES 162

List of Tables

Table Title Page

31 Treatment plan 46 32 Different treatments used in the biological study 54 41 Chemical composition of barley flour 57 42 Chemical Analysis of β-glucan 59 43 Mean sum of squares for color values (L a b) of stored β-glucan

beverages 62

44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

63

45 Effect of treatments and storage intervals on the a-value of stored β-glucan beverages

66

46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

68

47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

71

48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

72

49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

75

410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

76

411 Mean sum of squares for pH acidity and ascorbic acid content of stored beverages

78

412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

79

413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

82

414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

84

415 Mean sum of squares for reducing non reducing and total sugar content of stored beverages

87

416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

88

417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

90

418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

92

Table Title Page

419 Effect of treatments and storage intervals on the total plate count

(CFUml) of stored β-glucan beverages 94

420 Mean sum of squares for sensory evaluation of stored beverages 96 421 Effect of treatments and storage intervals on the color score of

stored β-glucan beverages 97

422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

100

423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

103

424 Effect of treatments and storage intervals and on the sourness score of stored β-glucan beverages

105

425 Effect of storage intervals and treatments on the overall acceptability score of stored β-glucan beverages

108

426 Mean sum of squares for blood lipid profile of volunteers 110 427 Effect of β-glucan supplemented beverage on serum total

cholesterol content (mgdl) of healthy subjects 111

428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

115

429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

119

430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

123

431 Mean sum of squares for blood glucose contents of volunteers 127 432 Effect of β-glucan beverage on blood glucose (mgdl) content of

with different time intervals 127

433 Interactive effect of diets and time scale intervals on the blood glucose contents (mgdl) of volunteers

127

434 Interactive effect of diets and study duration on the blood glucose contents (mgdl) of volunteers

128

List of Figures

Fig Title Page

31 Preparation of β -glucan beverage 47 41 Percent decrease in the serum total cholesterol level of subjects fed

on different beverages 111

42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of healthy volunteers

112

43 Percent decrease in the serum triglycerides level of subjects fed on different beverages

115

44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy volunteers

116

45 Percent decrease in the serum LDL level of subjects fed on different beverages

119

46 Effect of β-glucan beverage on LDL (mgdl) content of healthy volunteers

120

47 Percent increase in the serum HDL level of subjects fed on different beverages

123

48 Effect of β-glucan beverage on HDL (mgdl) content of healthy volunteers

124

49 Effect of β-glucan beverage on blood glucose (mgdl) content of healthy volunteers

128

List of Appendices

Appendix Title Page

I Composit ion of functional beverage 162

II 9 Point Hedonic Scale 163

III Food frequency questionnaire 164

IV Demographic information performa (subjects) 165

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

LITERATURE CITED

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Aastrup S 1979a The effect of rain on β -glucan content in barley grains Carlsberg esearch Communications 44381-393

Aditya K T Yokota S Suzuki and H Etoh 2008 Sub crit ical Water Extraction of Barley to Produce a Functional Drink

Biosci Biotechnol Biochem 72(1)236-239

AERI 1896 The Agricultural Economics Research Institute Balance Sheet for Food Commodities Finland 1985 The Insti tute Helsinki

Akubor PI 2003 Influence of storage on the physicochemical microbiological and sensory properties of heat and chemically treated melon-banana beverage Plant Foods for Human Nutri 58 1ndash10

Alessandra DC P Antonio V Vincenzo A Mario 2004 Changes of f lavonoids vitamin C and antioxidant capacity in minimally processed citrus segments and juices during storage Food Chem 84 99-105

Aman P H Graham AC Til ly 1989 Content and solubil i ty of mixed-l inked (1-3) (1-4)- β -D-glucan in barley and oats during kernel development and storage J Cereal Sci 1045-50

Anderson J W 1980 Dietary f iber and diabetes in Medical Aspects of Dietry Fiber G A spil ler and R M Key eds Plenum Medical Book Company New York

Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

Anderson J W DB Spencer CC Hamilton SF Smith and J Tietyen CA Bryant P Oeltgen 1990 Oat-bran cereal lowers serum total and LDL cholesterol in hypercholesterolemic men Am J Clin Nutri 52 495-499

139

Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

Annoni G BM Botasso D Ciaci MF Donato and A Tripodi 1982 Liquid tr iglycerides (GPO-PAP) Medi Diagnostic I taly Lab J Res Lab Med 9 115-116

AOAC 2000 Official Methods of Analysis The Association of the Official Analytical Chemists 20 t h Ed Arlington USA

Arndt EA 2006 Whole-grain barley for todays health and wellness needs ConAgra Foods Inc Omaha NE 51(1) 20-22

Assmann G 1979 HDL-cholesterol precipitant Randox Labs Ltd CrumLin Co Antrim N Ireland Internist 20559-567

Babsky NE J L Toribio and J E Lozano 1986 Influence of storage on the composit ion of clarif ied apple juice concentrate J Food Sci 51 (3) 564-67

Ballance GM WOS Meredith 1976 Purif ication and partial characterization of an endo- β -13-glucanase from green malt J Inst Brew 8264-67

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Bansema C 2000 Development of a barley P-glucan beverage with and without whey protein Isolate MSc thesis Edmonton Alberta Canada

Basman A and HK Ksel 1999 Properties and composit ion of Turkish f lat bread (bazlama) supplemented with barley f lour and wheat bran Cereal Chem 76506ndash511

Beer MU E Arrigoni and R Amado 1995 Effect of oat gum on blood cholesterol levels in healthy young men Europ J Clin Nutri 49517ndash522

140

Beer MU PJ Wood J Weisz N Fi l l ion 1997 Effect of cooking and storage on the amount and molecular weight of (1rarr3) (1rarr4) - β -D-glucan extracted from oat products by an in vitro digestion system Cereal Chem 74 705-709

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 A Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bender DA and AE Bender 1999 Bendersrsquo Dictionary of Nutrit ion and Food Technology 7 t h ed Woodhead Publishing Abington

Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

Berglund PT CE Fastnaught ET Holm 1992 Food uses of waxy hull- less barley Cereal Foods World 37707ndash714

Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

Bhatty RS 1992 Total and extractable β -glucan contents of oats and their relationship to viscosity J Cer Sci 15185-192

Bhatty RS 1995 Laboratory and pilot plant extraction and purif ication of b-glucans from hull- less barley and oat bran J Cer Sci 22163ndash170

Bhatty RS 1996 Production of food malt from hull- less barley Cereal Chem 73(1) 75-80

Bhatty RS AW MacGregor and BG Rossnagel 1991 Total and acid-soluble β -glucan content of hulless barley and its relationship to acid-extract viscosity Cereal Chem 68221-227

Bhatty RS1986 Physiochemical and Functional (Breadmaking) Properties of Hull- less Barley Fractions Cereal Chem 6331-35

141

Bibek R 2001 Fundamental Food Microbiology 2nd edn The CRC press Ltd Washington DC pp 56-90

Bingham SA NE Day R Luben P Ferrari N Sl imani T Norat F Lavel E Kesse A Nieters H Boeing A Tjoslashnneland K Overvad C Martinez M Dorrensoro CA Gonzalez TJ Key A Trichopoulou A Naska P Vineis R Tumino V Krogh HB Bueno-de-Mesquita PHM Peeters G Berglung G Hallmans E Lund G Skele R Kaaks and E Riboli 2003 Dietary f ibre in food and protection against colorectal cancer in the European Prospective Investigation into Cancer and Nutrit ion (EPIC) an observational study Lancet 3611496-501

Bioumlrklund M A van Rees RP Mensink and G Oumlnning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucans from oats and barley a randomised dose-controlled tr ial Eur J Clin Nutri 591272-1281

Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

Bjorck I AC Eliasson A Drews M Gudmundsson and R Karlsson 1990 some nutrit ional properties of starch and dietary f iber in barley genotypes containing different levels of amylose Cereal Chem 67 327

BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

Brand J S Colagiuri S Crossman A Allen D Roberts and S Truswell 1991 Low-glycemic index foods improve long term glycemic control in NIDDM Diabetes Care 14 95ndash101

142

Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

Brennan CS and LJ Cleary 2005 The potential use of cereal (1314)- β -D-glucans as functional food ingredients J Cer Sci 421ndash13

Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

Brown L B Rosner W Willet and FM Sacks 1999 Cholesterol lowering effects of dietary f iber a meta analysis Am J Clin Nutri 69 (1) 30 42

Brunswick P DJ Manner and J K Stark 1987 Development of β -D-glucanases during germination of barley and the effect of ki lning on individual isoenzymes J Inst Brew 93181-186

Bryan D J Robert AT Wilson T Carlson S Frazer GH Zheng 2003 β -Glucan Fractions from Barley and Oats Are Similarly Antiatherogenic in Hypercholesterolemic Syrian Golden Hamsters The American Society for Nutrit ional Sciences J Nutri Metabolism 133468-475

Buliga GS DA Brant and GB Fincher 1986 The sequence statist ics and solution configration of barley (1rarr3) (1rarr4) - β -D-glucan Carbohydr Res 57139-156

Burkus Z 1996 Barley P-Glucan Extraction Functional Properties and Interactions with Food Components MSc thesis Edmonton AlbertaCanda

Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloid F Glicksman M (Ed) p 49-93 CRC Press Inc Boca Raton

Burkus Z 1996 Barley β -glucan Extraction Functional properties and interaction with food components MSc Thesis Dept of Agricultural Food and Nutrit ional Science Univ of Alberta Edmonton Canada

143

Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

Burkus Z and F Temell i 1999 Glucan concentrate J Food Sci 64198-201 Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloidr Glicksman M (Ed) p 49-93 CRC Press hc Boca Raton FL

Burkus Z and F Temell i 2005 Rheological properties of barley β -glucan Carbohydr Polym 59 459ndash465

Burkus Z F Temell i 1999 Gelation of barley β -glucan - concentrate J Food Sci 64198-201

Calix FD and N Bardrie 2004 Consumer acceptance and physicochemical quality of processed red sorrelroselle (Hibiscus sabdar i f fa L) sauces from enzymatic extracted calyces 4 141-148

Carpita NC 1996 Structure and biogenesis of cel l walls of grasses Annual Rev Plant Physiol Plat Molecular Biol 47445-476

Carr J M S Glatter J L Jeraci and B A Lewis 1990 Enzymes Determination of Beta-Glucan in Cereal-Based Food Products Cereal Chem 67226-229

Casterl ine J L CJ Oles and Y Ku 1997 In vitro fermentation of various food f iber reactions J Agric Food Chem 452463ndash2467

Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

Chowdhury MGF MN Islam MS Is lam T Is lam and MS Hossain 2008 Study on Preparation and Shelf-Life of Mixed Juice Based on Wood Apple and Papaya J Soil Nature 2(3) 50-60

Chung OK and Y Pomeranz 1985 Amino acids in cereal proteins and protein fractions Ch 5 in Digesfibi l i~ and

144

Amino Acid Availabil i ty in Cereals andOilseeds J W Finley and DT Hopkins (Eds) pp 169-232 AACC St Paul MN

Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

Crandall PG CS Chen and KC Davis 1987 Preparation and storage of 72 brix orange juice concentration J Food Sci 52 (3) 381

Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

Daw ZY YSA El-Gizaw and AMB Said 1994 Microbiological evaluation of some local juices and drinks Chemie Mikrobiologie Technologie der Lebensmittel 168ndash15

Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

Dawkins NL I A Nnanna 1993 Studies on oat gum [(1rarr31rarr4)- β-D-glucan] Composit ion molecular weight est imation and rheological properties Food Hydrocol 9 1-7

Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

Dohnalek MH 2004 The role of f ibre in cl inical nutrit ion In Van der Kamp JW Asp NG Miller J J Schaafsma G (Ed) Dietary f ibre bioactive carbohydrates for food and feed Wageningen Academic Publishers Wageningen pp 271294

145

Dongowski G M Huth E Gebhardt and W Flamme 2002 Dietary f iber-rich barley products beneficial ly affect the intestinal tract of rats J Nutri 132(12) 3704-14

Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

Falade OS OR Sowunmi A Oladipo A Tobosun and SRA Adewusi 2003 The level of organic acids in some Nigerian fruit and their effect on mineral availabil i ty in composite diet Pak J Nutri 2(2) 82-83

Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

FDA 1996 Food labeling Health claims oats and coronary heart disease Federal Register 61 (3) January 4

Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

Frazier WC and EM Foster 1958 Laboratory Manual for Food Microbiology Burgess Pub Co Minneapolis Minnisota USA

Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

146

Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

Giese J H 1992 Hitt ing the spot Beverages and beverage technology Food Technol 4670-72 74-75 78-80

Godara RK and OP Pareek 1985 Effect of temperature in storage of ready to serve date juice beverages indian j agric Sci 55 (5) 347-349 (FSTA 18 (4) 78 1986)

Gonzalez ER and S Leeson 2000 An investigation on the preservation of kununndashzaki an African fermented cereal based food drink Acta Alimentaria 29 385ndash92

GOP 2008 Government of Pakistan Finance Division Economic Advisor s Wing Islamabad Pakistan

Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

Hallfr isch J DJ Schofield KM Behall 2003 Physiological responses of men and women to barley and oat extracts (NutrimX) I I Comparison of glucose and insulin responses Cereal Chem 8080ndash83

Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

147

Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

Lawless HT and H heymann Sensory evaluation of food Principles and Practices Gaithersburg MD Aspen Publishers ISSN 1572-0330) Oorspr uitg New York [etc ] Chapman amp Hall 1998

Hashimoto S MD Shogren Y Pomeranz 1987 Cereal Pentosans Their est imation and signif icance I Pentosans in wheat and milled wheat products Cereal Chem 64(1) 30-34

Hassan SA 1976 Effect of storage on physico-chemical characterist ics of carbonated orange juice Msc thesis Food Tech Deptt WPAU Lyallpur

Hatcher WSJ R J L Weihe DF Split tstoesser EC Hil l and ME Parish 1992 Fruit Beverages In Compendium of methods for the microbiological examination of foods Vanderzant C Split tstoesser DF (eds) American Public Health Association Washington DC

Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

Hil l M J and FR Path 1998 Cereals dietary f iber and cancer Nutri Res 18563ndash659

Hil l iam M 2000 Functional foodndashndashHow big is the market The World of Food Ingredients 12 50ndash2

Holsinger V H LP Posati and ED DeVilbiss 1974 Whey beverages a review J Dairy Sci 57(7) 849ndash859

Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

148

47ndash82) St Paul MN USA American Association of Cereal Chemists

Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

Jadhav SJ S E Lutz VM Ghorpade and DK Salunkhe 1998 Barley chemistry and value-added processing Crit ical Rev Food Sci 3823ndash171

Jal i l i T REC Wildman DM Medeiros 2000 Nutraceutical roles of dietary f iber J Nutraceutical functional and Medi foods 2 19-34

Jansen MC HB Bueno-de-Mesquita R Buzina F Fidanza A Menotti H Blackburn AM Nissinen FJ Kok D Kromhout 1999 Dietary f iber and plant foods in relation to colorectal cancer mortal i ty The Seven Countries Study Inter J Canc 81 174-179

Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high

CC OO NN TT EE NN TT SS

CHAPTER TITLE PAGE

ABSTRACT

1 INTRODUCTION 1

2 REVIEW OF LITERATURE 6

3 MATERIALS AND METHODS 39

4 RESULTS AND DISCUSSION 56

5 SUMMARY 131

CONCLUSIONS 136

RECOMMENDATIONS 137

LITERATURE CITED 138

APPENDICES 162

List of Tables

Table Title Page

31 Treatment plan 46 32 Different treatments used in the biological study 54 41 Chemical composition of barley flour 57 42 Chemical Analysis of β-glucan 59 43 Mean sum of squares for color values (L a b) of stored β-glucan

beverages 62

44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

63

45 Effect of treatments and storage intervals on the a-value of stored β-glucan beverages

66

46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

68

47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

71

48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

72

49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

75

410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

76

411 Mean sum of squares for pH acidity and ascorbic acid content of stored beverages

78

412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

79

413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

82

414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

84

415 Mean sum of squares for reducing non reducing and total sugar content of stored beverages

87

416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

88

417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

90

418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

92

Table Title Page

419 Effect of treatments and storage intervals on the total plate count

(CFUml) of stored β-glucan beverages 94

420 Mean sum of squares for sensory evaluation of stored beverages 96 421 Effect of treatments and storage intervals on the color score of

stored β-glucan beverages 97

422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

100

423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

103

424 Effect of treatments and storage intervals and on the sourness score of stored β-glucan beverages

105

425 Effect of storage intervals and treatments on the overall acceptability score of stored β-glucan beverages

108

426 Mean sum of squares for blood lipid profile of volunteers 110 427 Effect of β-glucan supplemented beverage on serum total

cholesterol content (mgdl) of healthy subjects 111

428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

115

429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

119

430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

123

431 Mean sum of squares for blood glucose contents of volunteers 127 432 Effect of β-glucan beverage on blood glucose (mgdl) content of

with different time intervals 127

433 Interactive effect of diets and time scale intervals on the blood glucose contents (mgdl) of volunteers

127

434 Interactive effect of diets and study duration on the blood glucose contents (mgdl) of volunteers

128

List of Figures

Fig Title Page

31 Preparation of β -glucan beverage 47 41 Percent decrease in the serum total cholesterol level of subjects fed

on different beverages 111

42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of healthy volunteers

112

43 Percent decrease in the serum triglycerides level of subjects fed on different beverages

115

44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy volunteers

116

45 Percent decrease in the serum LDL level of subjects fed on different beverages

119

46 Effect of β-glucan beverage on LDL (mgdl) content of healthy volunteers

120

47 Percent increase in the serum HDL level of subjects fed on different beverages

123

48 Effect of β-glucan beverage on HDL (mgdl) content of healthy volunteers

124

49 Effect of β-glucan beverage on blood glucose (mgdl) content of healthy volunteers

128

List of Appendices

Appendix Title Page

I Composit ion of functional beverage 162

II 9 Point Hedonic Scale 163

III Food frequency questionnaire 164

IV Demographic information performa (subjects) 165

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

LITERATURE CITED

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Aastrup S 1979a The effect of rain on β -glucan content in barley grains Carlsberg esearch Communications 44381-393

Aditya K T Yokota S Suzuki and H Etoh 2008 Sub crit ical Water Extraction of Barley to Produce a Functional Drink

Biosci Biotechnol Biochem 72(1)236-239

AERI 1896 The Agricultural Economics Research Institute Balance Sheet for Food Commodities Finland 1985 The Insti tute Helsinki

Akubor PI 2003 Influence of storage on the physicochemical microbiological and sensory properties of heat and chemically treated melon-banana beverage Plant Foods for Human Nutri 58 1ndash10

Alessandra DC P Antonio V Vincenzo A Mario 2004 Changes of f lavonoids vitamin C and antioxidant capacity in minimally processed citrus segments and juices during storage Food Chem 84 99-105

Aman P H Graham AC Til ly 1989 Content and solubil i ty of mixed-l inked (1-3) (1-4)- β -D-glucan in barley and oats during kernel development and storage J Cereal Sci 1045-50

Anderson J W 1980 Dietary f iber and diabetes in Medical Aspects of Dietry Fiber G A spil ler and R M Key eds Plenum Medical Book Company New York

Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

Anderson J W DB Spencer CC Hamilton SF Smith and J Tietyen CA Bryant P Oeltgen 1990 Oat-bran cereal lowers serum total and LDL cholesterol in hypercholesterolemic men Am J Clin Nutri 52 495-499

139

Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

Annoni G BM Botasso D Ciaci MF Donato and A Tripodi 1982 Liquid tr iglycerides (GPO-PAP) Medi Diagnostic I taly Lab J Res Lab Med 9 115-116

AOAC 2000 Official Methods of Analysis The Association of the Official Analytical Chemists 20 t h Ed Arlington USA

Arndt EA 2006 Whole-grain barley for todays health and wellness needs ConAgra Foods Inc Omaha NE 51(1) 20-22

Assmann G 1979 HDL-cholesterol precipitant Randox Labs Ltd CrumLin Co Antrim N Ireland Internist 20559-567

Babsky NE J L Toribio and J E Lozano 1986 Influence of storage on the composit ion of clarif ied apple juice concentrate J Food Sci 51 (3) 564-67

Ballance GM WOS Meredith 1976 Purif ication and partial characterization of an endo- β -13-glucanase from green malt J Inst Brew 8264-67

Bamforth CW and AHP Barclay 1993 Malting technology and the uses of malt In Barley Chemistry and Technology (eds AW MacGregor and RS Bhatty) by Am Assoc Cereal Chem St Paul USA pp 297-354

Bansema C 2000 Development of a barley P-glucan beverage with and without whey protein Isolate MSc thesis Edmonton Alberta Canada

Basman A and HK Ksel 1999 Properties and composit ion of Turkish f lat bread (bazlama) supplemented with barley f lour and wheat bran Cereal Chem 76506ndash511

Beer MU E Arrigoni and R Amado 1995 Effect of oat gum on blood cholesterol levels in healthy young men Europ J Clin Nutri 49517ndash522

140

Beer MU PJ Wood J Weisz N Fi l l ion 1997 Effect of cooking and storage on the amount and molecular weight of (1rarr3) (1rarr4) - β -D-glucan extracted from oat products by an in vitro digestion system Cereal Chem 74 705-709

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 A Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bender DA and AE Bender 1999 Bendersrsquo Dictionary of Nutrit ion and Food Technology 7 t h ed Woodhead Publishing Abington

Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

Berglund PT CE Fastnaught ET Holm 1992 Food uses of waxy hull- less barley Cereal Foods World 37707ndash714

Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

Bhatty RS 1992 Total and extractable β -glucan contents of oats and their relationship to viscosity J Cer Sci 15185-192

Bhatty RS 1995 Laboratory and pilot plant extraction and purif ication of b-glucans from hull- less barley and oat bran J Cer Sci 22163ndash170

Bhatty RS 1996 Production of food malt from hull- less barley Cereal Chem 73(1) 75-80

Bhatty RS AW MacGregor and BG Rossnagel 1991 Total and acid-soluble β -glucan content of hulless barley and its relationship to acid-extract viscosity Cereal Chem 68221-227

Bhatty RS1986 Physiochemical and Functional (Breadmaking) Properties of Hull- less Barley Fractions Cereal Chem 6331-35

141

Bibek R 2001 Fundamental Food Microbiology 2nd edn The CRC press Ltd Washington DC pp 56-90

Bingham SA NE Day R Luben P Ferrari N Sl imani T Norat F Lavel E Kesse A Nieters H Boeing A Tjoslashnneland K Overvad C Martinez M Dorrensoro CA Gonzalez TJ Key A Trichopoulou A Naska P Vineis R Tumino V Krogh HB Bueno-de-Mesquita PHM Peeters G Berglung G Hallmans E Lund G Skele R Kaaks and E Riboli 2003 Dietary f ibre in food and protection against colorectal cancer in the European Prospective Investigation into Cancer and Nutrit ion (EPIC) an observational study Lancet 3611496-501

Bioumlrklund M A van Rees RP Mensink and G Oumlnning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucans from oats and barley a randomised dose-controlled tr ial Eur J Clin Nutri 591272-1281

Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

Bjorck I AC Eliasson A Drews M Gudmundsson and R Karlsson 1990 some nutrit ional properties of starch and dietary f iber in barley genotypes containing different levels of amylose Cereal Chem 67 327

BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

Brand J S Colagiuri S Crossman A Allen D Roberts and S Truswell 1991 Low-glycemic index foods improve long term glycemic control in NIDDM Diabetes Care 14 95ndash101

142

Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

Brennan CS and LJ Cleary 2005 The potential use of cereal (1314)- β -D-glucans as functional food ingredients J Cer Sci 421ndash13

Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

Brown L B Rosner W Willet and FM Sacks 1999 Cholesterol lowering effects of dietary f iber a meta analysis Am J Clin Nutri 69 (1) 30 42

Brunswick P DJ Manner and J K Stark 1987 Development of β -D-glucanases during germination of barley and the effect of ki lning on individual isoenzymes J Inst Brew 93181-186

Bryan D J Robert AT Wilson T Carlson S Frazer GH Zheng 2003 β -Glucan Fractions from Barley and Oats Are Similarly Antiatherogenic in Hypercholesterolemic Syrian Golden Hamsters The American Society for Nutrit ional Sciences J Nutri Metabolism 133468-475

Buliga GS DA Brant and GB Fincher 1986 The sequence statist ics and solution configration of barley (1rarr3) (1rarr4) - β -D-glucan Carbohydr Res 57139-156

Burkus Z 1996 Barley P-Glucan Extraction Functional Properties and Interactions with Food Components MSc thesis Edmonton AlbertaCanda

Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloid F Glicksman M (Ed) p 49-93 CRC Press Inc Boca Raton

Burkus Z 1996 Barley β -glucan Extraction Functional properties and interaction with food components MSc Thesis Dept of Agricultural Food and Nutrit ional Science Univ of Alberta Edmonton Canada

143

Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

Burkus Z and F Temell i 1999 Glucan concentrate J Food Sci 64198-201 Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloidr Glicksman M (Ed) p 49-93 CRC Press hc Boca Raton FL

Burkus Z and F Temell i 2005 Rheological properties of barley β -glucan Carbohydr Polym 59 459ndash465

Burkus Z F Temell i 1999 Gelation of barley β -glucan - concentrate J Food Sci 64198-201

Calix FD and N Bardrie 2004 Consumer acceptance and physicochemical quality of processed red sorrelroselle (Hibiscus sabdar i f fa L) sauces from enzymatic extracted calyces 4 141-148

Carpita NC 1996 Structure and biogenesis of cel l walls of grasses Annual Rev Plant Physiol Plat Molecular Biol 47445-476

Carr J M S Glatter J L Jeraci and B A Lewis 1990 Enzymes Determination of Beta-Glucan in Cereal-Based Food Products Cereal Chem 67226-229

Casterl ine J L CJ Oles and Y Ku 1997 In vitro fermentation of various food f iber reactions J Agric Food Chem 452463ndash2467

Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

Chowdhury MGF MN Islam MS Is lam T Is lam and MS Hossain 2008 Study on Preparation and Shelf-Life of Mixed Juice Based on Wood Apple and Papaya J Soil Nature 2(3) 50-60

Chung OK and Y Pomeranz 1985 Amino acids in cereal proteins and protein fractions Ch 5 in Digesfibi l i~ and

144

Amino Acid Availabil i ty in Cereals andOilseeds J W Finley and DT Hopkins (Eds) pp 169-232 AACC St Paul MN

Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

Crandall PG CS Chen and KC Davis 1987 Preparation and storage of 72 brix orange juice concentration J Food Sci 52 (3) 381

Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

Daw ZY YSA El-Gizaw and AMB Said 1994 Microbiological evaluation of some local juices and drinks Chemie Mikrobiologie Technologie der Lebensmittel 168ndash15

Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

Dawkins NL I A Nnanna 1993 Studies on oat gum [(1rarr31rarr4)- β-D-glucan] Composit ion molecular weight est imation and rheological properties Food Hydrocol 9 1-7

Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

Dohnalek MH 2004 The role of f ibre in cl inical nutrit ion In Van der Kamp JW Asp NG Miller J J Schaafsma G (Ed) Dietary f ibre bioactive carbohydrates for food and feed Wageningen Academic Publishers Wageningen pp 271294

145

Dongowski G M Huth E Gebhardt and W Flamme 2002 Dietary f iber-rich barley products beneficial ly affect the intestinal tract of rats J Nutri 132(12) 3704-14

Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

Falade OS OR Sowunmi A Oladipo A Tobosun and SRA Adewusi 2003 The level of organic acids in some Nigerian fruit and their effect on mineral availabil i ty in composite diet Pak J Nutri 2(2) 82-83

Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

FDA 1996 Food labeling Health claims oats and coronary heart disease Federal Register 61 (3) January 4

Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

Frazier WC and EM Foster 1958 Laboratory Manual for Food Microbiology Burgess Pub Co Minneapolis Minnisota USA

Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

146

Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

Giese J H 1992 Hitt ing the spot Beverages and beverage technology Food Technol 4670-72 74-75 78-80

Godara RK and OP Pareek 1985 Effect of temperature in storage of ready to serve date juice beverages indian j agric Sci 55 (5) 347-349 (FSTA 18 (4) 78 1986)

Gonzalez ER and S Leeson 2000 An investigation on the preservation of kununndashzaki an African fermented cereal based food drink Acta Alimentaria 29 385ndash92

GOP 2008 Government of Pakistan Finance Division Economic Advisor s Wing Islamabad Pakistan

Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

Hallfr isch J DJ Schofield KM Behall 2003 Physiological responses of men and women to barley and oat extracts (NutrimX) I I Comparison of glucose and insulin responses Cereal Chem 8080ndash83

Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

147

Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

Lawless HT and H heymann Sensory evaluation of food Principles and Practices Gaithersburg MD Aspen Publishers ISSN 1572-0330) Oorspr uitg New York [etc ] Chapman amp Hall 1998

Hashimoto S MD Shogren Y Pomeranz 1987 Cereal Pentosans Their est imation and signif icance I Pentosans in wheat and milled wheat products Cereal Chem 64(1) 30-34

Hassan SA 1976 Effect of storage on physico-chemical characterist ics of carbonated orange juice Msc thesis Food Tech Deptt WPAU Lyallpur

Hatcher WSJ R J L Weihe DF Split tstoesser EC Hil l and ME Parish 1992 Fruit Beverages In Compendium of methods for the microbiological examination of foods Vanderzant C Split tstoesser DF (eds) American Public Health Association Washington DC

Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

Hil l M J and FR Path 1998 Cereals dietary f iber and cancer Nutri Res 18563ndash659

Hil l iam M 2000 Functional foodndashndashHow big is the market The World of Food Ingredients 12 50ndash2

Holsinger V H LP Posati and ED DeVilbiss 1974 Whey beverages a review J Dairy Sci 57(7) 849ndash859

Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

148

47ndash82) St Paul MN USA American Association of Cereal Chemists

Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

Jadhav SJ S E Lutz VM Ghorpade and DK Salunkhe 1998 Barley chemistry and value-added processing Crit ical Rev Food Sci 3823ndash171

Jal i l i T REC Wildman DM Medeiros 2000 Nutraceutical roles of dietary f iber J Nutraceutical functional and Medi foods 2 19-34

Jansen MC HB Bueno-de-Mesquita R Buzina F Fidanza A Menotti H Blackburn AM Nissinen FJ Kok D Kromhout 1999 Dietary f iber and plant foods in relation to colorectal cancer mortal i ty The Seven Countries Study Inter J Canc 81 174-179

Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high

List of Tables

Table Title Page

31 Treatment plan 46 32 Different treatments used in the biological study 54 41 Chemical composition of barley flour 57 42 Chemical Analysis of β-glucan 59 43 Mean sum of squares for color values (L a b) of stored β-glucan

beverages 62

44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

63

45 Effect of treatments and storage intervals on the a-value of stored β-glucan beverages

66

46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

68

47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

71

48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

72

49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

75

410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

76

411 Mean sum of squares for pH acidity and ascorbic acid content of stored beverages

78

412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

79

413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

82

414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

84

415 Mean sum of squares for reducing non reducing and total sugar content of stored beverages

87

416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

88

417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

90

418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

92

Table Title Page

419 Effect of treatments and storage intervals on the total plate count

(CFUml) of stored β-glucan beverages 94

420 Mean sum of squares for sensory evaluation of stored beverages 96 421 Effect of treatments and storage intervals on the color score of

stored β-glucan beverages 97

422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

100

423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

103

424 Effect of treatments and storage intervals and on the sourness score of stored β-glucan beverages

105

425 Effect of storage intervals and treatments on the overall acceptability score of stored β-glucan beverages

108

426 Mean sum of squares for blood lipid profile of volunteers 110 427 Effect of β-glucan supplemented beverage on serum total

cholesterol content (mgdl) of healthy subjects 111

428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

115

429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

119

430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

123

431 Mean sum of squares for blood glucose contents of volunteers 127 432 Effect of β-glucan beverage on blood glucose (mgdl) content of

with different time intervals 127

433 Interactive effect of diets and time scale intervals on the blood glucose contents (mgdl) of volunteers

127

434 Interactive effect of diets and study duration on the blood glucose contents (mgdl) of volunteers

128

List of Figures

Fig Title Page

31 Preparation of β -glucan beverage 47 41 Percent decrease in the serum total cholesterol level of subjects fed

on different beverages 111

42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of healthy volunteers

112

43 Percent decrease in the serum triglycerides level of subjects fed on different beverages

115

44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy volunteers

116

45 Percent decrease in the serum LDL level of subjects fed on different beverages

119

46 Effect of β-glucan beverage on LDL (mgdl) content of healthy volunteers

120

47 Percent increase in the serum HDL level of subjects fed on different beverages

123

48 Effect of β-glucan beverage on HDL (mgdl) content of healthy volunteers

124

49 Effect of β-glucan beverage on blood glucose (mgdl) content of healthy volunteers

128

List of Appendices

Appendix Title Page

I Composit ion of functional beverage 162

II 9 Point Hedonic Scale 163

III Food frequency questionnaire 164

IV Demographic information performa (subjects) 165

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

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Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

Anderson J W DB Spencer CC Hamilton SF Smith and J Tietyen CA Bryant P Oeltgen 1990 Oat-bran cereal lowers serum total and LDL cholesterol in hypercholesterolemic men Am J Clin Nutri 52 495-499

139

Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

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Beer MU E Arrigoni and R Amado 1995 Effect of oat gum on blood cholesterol levels in healthy young men Europ J Clin Nutri 49517ndash522

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Beer MU PJ Wood J Weisz N Fi l l ion 1997 Effect of cooking and storage on the amount and molecular weight of (1rarr3) (1rarr4) - β -D-glucan extracted from oat products by an in vitro digestion system Cereal Chem 74 705-709

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

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Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

Berglund PT CE Fastnaught ET Holm 1992 Food uses of waxy hull- less barley Cereal Foods World 37707ndash714

Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

Bhatty RS 1992 Total and extractable β -glucan contents of oats and their relationship to viscosity J Cer Sci 15185-192

Bhatty RS 1995 Laboratory and pilot plant extraction and purif ication of b-glucans from hull- less barley and oat bran J Cer Sci 22163ndash170

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Bioumlrklund M A van Rees RP Mensink and G Oumlnning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucans from oats and barley a randomised dose-controlled tr ial Eur J Clin Nutri 591272-1281

Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

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BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

Brand J S Colagiuri S Crossman A Allen D Roberts and S Truswell 1991 Low-glycemic index foods improve long term glycemic control in NIDDM Diabetes Care 14 95ndash101

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Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

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Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

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Bryan D J Robert AT Wilson T Carlson S Frazer GH Zheng 2003 β -Glucan Fractions from Barley and Oats Are Similarly Antiatherogenic in Hypercholesterolemic Syrian Golden Hamsters The American Society for Nutrit ional Sciences J Nutri Metabolism 133468-475

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Burkus Z 1996 Barley P-Glucan Extraction Functional Properties and Interactions with Food Components MSc thesis Edmonton AlbertaCanda

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Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

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Calix FD and N Bardrie 2004 Consumer acceptance and physicochemical quality of processed red sorrelroselle (Hibiscus sabdar i f fa L) sauces from enzymatic extracted calyces 4 141-148

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Carr J M S Glatter J L Jeraci and B A Lewis 1990 Enzymes Determination of Beta-Glucan in Cereal-Based Food Products Cereal Chem 67226-229

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Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

Chowdhury MGF MN Islam MS Is lam T Is lam and MS Hossain 2008 Study on Preparation and Shelf-Life of Mixed Juice Based on Wood Apple and Papaya J Soil Nature 2(3) 50-60

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Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

Crandall PG CS Chen and KC Davis 1987 Preparation and storage of 72 brix orange juice concentration J Food Sci 52 (3) 381

Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

Daw ZY YSA El-Gizaw and AMB Said 1994 Microbiological evaluation of some local juices and drinks Chemie Mikrobiologie Technologie der Lebensmittel 168ndash15

Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

Dawkins NL I A Nnanna 1993 Studies on oat gum [(1rarr31rarr4)- β-D-glucan] Composit ion molecular weight est imation and rheological properties Food Hydrocol 9 1-7

Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

Dohnalek MH 2004 The role of f ibre in cl inical nutrit ion In Van der Kamp JW Asp NG Miller J J Schaafsma G (Ed) Dietary f ibre bioactive carbohydrates for food and feed Wageningen Academic Publishers Wageningen pp 271294

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Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

Falade OS OR Sowunmi A Oladipo A Tobosun and SRA Adewusi 2003 The level of organic acids in some Nigerian fruit and their effect on mineral availabil i ty in composite diet Pak J Nutri 2(2) 82-83

Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

FDA 1996 Food labeling Health claims oats and coronary heart disease Federal Register 61 (3) January 4

Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

Frazier WC and EM Foster 1958 Laboratory Manual for Food Microbiology Burgess Pub Co Minneapolis Minnisota USA

Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

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Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

Giese J H 1992 Hitt ing the spot Beverages and beverage technology Food Technol 4670-72 74-75 78-80

Godara RK and OP Pareek 1985 Effect of temperature in storage of ready to serve date juice beverages indian j agric Sci 55 (5) 347-349 (FSTA 18 (4) 78 1986)

Gonzalez ER and S Leeson 2000 An investigation on the preservation of kununndashzaki an African fermented cereal based food drink Acta Alimentaria 29 385ndash92

GOP 2008 Government of Pakistan Finance Division Economic Advisor s Wing Islamabad Pakistan

Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

Hallfr isch J DJ Schofield KM Behall 2003 Physiological responses of men and women to barley and oat extracts (NutrimX) I I Comparison of glucose and insulin responses Cereal Chem 8080ndash83

Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

147

Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

Lawless HT and H heymann Sensory evaluation of food Principles and Practices Gaithersburg MD Aspen Publishers ISSN 1572-0330) Oorspr uitg New York [etc ] Chapman amp Hall 1998

Hashimoto S MD Shogren Y Pomeranz 1987 Cereal Pentosans Their est imation and signif icance I Pentosans in wheat and milled wheat products Cereal Chem 64(1) 30-34

Hassan SA 1976 Effect of storage on physico-chemical characterist ics of carbonated orange juice Msc thesis Food Tech Deptt WPAU Lyallpur

Hatcher WSJ R J L Weihe DF Split tstoesser EC Hil l and ME Parish 1992 Fruit Beverages In Compendium of methods for the microbiological examination of foods Vanderzant C Split tstoesser DF (eds) American Public Health Association Washington DC

Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

Hil l M J and FR Path 1998 Cereals dietary f iber and cancer Nutri Res 18563ndash659

Hil l iam M 2000 Functional foodndashndashHow big is the market The World of Food Ingredients 12 50ndash2

Holsinger V H LP Posati and ED DeVilbiss 1974 Whey beverages a review J Dairy Sci 57(7) 849ndash859

Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

148

47ndash82) St Paul MN USA American Association of Cereal Chemists

Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

Jadhav SJ S E Lutz VM Ghorpade and DK Salunkhe 1998 Barley chemistry and value-added processing Crit ical Rev Food Sci 3823ndash171

Jal i l i T REC Wildman DM Medeiros 2000 Nutraceutical roles of dietary f iber J Nutraceutical functional and Medi foods 2 19-34

Jansen MC HB Bueno-de-Mesquita R Buzina F Fidanza A Menotti H Blackburn AM Nissinen FJ Kok D Kromhout 1999 Dietary f iber and plant foods in relation to colorectal cancer mortal i ty The Seven Countries Study Inter J Canc 81 174-179

Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high

Table Title Page

419 Effect of treatments and storage intervals on the total plate count

(CFUml) of stored β-glucan beverages 94

420 Mean sum of squares for sensory evaluation of stored beverages 96 421 Effect of treatments and storage intervals on the color score of

stored β-glucan beverages 97

422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

100

423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

103

424 Effect of treatments and storage intervals and on the sourness score of stored β-glucan beverages

105

425 Effect of storage intervals and treatments on the overall acceptability score of stored β-glucan beverages

108

426 Mean sum of squares for blood lipid profile of volunteers 110 427 Effect of β-glucan supplemented beverage on serum total

cholesterol content (mgdl) of healthy subjects 111

428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

115

429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

119

430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

123

431 Mean sum of squares for blood glucose contents of volunteers 127 432 Effect of β-glucan beverage on blood glucose (mgdl) content of

with different time intervals 127

433 Interactive effect of diets and time scale intervals on the blood glucose contents (mgdl) of volunteers

127

434 Interactive effect of diets and study duration on the blood glucose contents (mgdl) of volunteers

128

List of Figures

Fig Title Page

31 Preparation of β -glucan beverage 47 41 Percent decrease in the serum total cholesterol level of subjects fed

on different beverages 111

42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of healthy volunteers

112

43 Percent decrease in the serum triglycerides level of subjects fed on different beverages

115

44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy volunteers

116

45 Percent decrease in the serum LDL level of subjects fed on different beverages

119

46 Effect of β-glucan beverage on LDL (mgdl) content of healthy volunteers

120

47 Percent increase in the serum HDL level of subjects fed on different beverages

123

48 Effect of β-glucan beverage on HDL (mgdl) content of healthy volunteers

124

49 Effect of β-glucan beverage on blood glucose (mgdl) content of healthy volunteers

128

List of Appendices

Appendix Title Page

I Composit ion of functional beverage 162

II 9 Point Hedonic Scale 163

III Food frequency questionnaire 164

IV Demographic information performa (subjects) 165

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

LITERATURE CITED

AACC 2000 Approved Methods of American Association of Cereal Chemists The American Association of Cereal Chemists Inc St Paul Minnesota USA

Aastrup S 1979a The effect of rain on β -glucan content in barley grains Carlsberg esearch Communications 44381-393

Aditya K T Yokota S Suzuki and H Etoh 2008 Sub crit ical Water Extraction of Barley to Produce a Functional Drink

Biosci Biotechnol Biochem 72(1)236-239

AERI 1896 The Agricultural Economics Research Institute Balance Sheet for Food Commodities Finland 1985 The Insti tute Helsinki

Akubor PI 2003 Influence of storage on the physicochemical microbiological and sensory properties of heat and chemically treated melon-banana beverage Plant Foods for Human Nutri 58 1ndash10

Alessandra DC P Antonio V Vincenzo A Mario 2004 Changes of f lavonoids vitamin C and antioxidant capacity in minimally processed citrus segments and juices during storage Food Chem 84 99-105

Aman P H Graham AC Til ly 1989 Content and solubil i ty of mixed-l inked (1-3) (1-4)- β -D-glucan in barley and oats during kernel development and storage J Cereal Sci 1045-50

Anderson J W 1980 Dietary f iber and diabetes in Medical Aspects of Dietry Fiber G A spil ler and R M Key eds Plenum Medical Book Company New York

Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

Anderson J W DB Spencer CC Hamilton SF Smith and J Tietyen CA Bryant P Oeltgen 1990 Oat-bran cereal lowers serum total and LDL cholesterol in hypercholesterolemic men Am J Clin Nutri 52 495-499

139

Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

Annoni G BM Botasso D Ciaci MF Donato and A Tripodi 1982 Liquid tr iglycerides (GPO-PAP) Medi Diagnostic I taly Lab J Res Lab Med 9 115-116

AOAC 2000 Official Methods of Analysis The Association of the Official Analytical Chemists 20 t h Ed Arlington USA

Arndt EA 2006 Whole-grain barley for todays health and wellness needs ConAgra Foods Inc Omaha NE 51(1) 20-22

Assmann G 1979 HDL-cholesterol precipitant Randox Labs Ltd CrumLin Co Antrim N Ireland Internist 20559-567

Babsky NE J L Toribio and J E Lozano 1986 Influence of storage on the composit ion of clarif ied apple juice concentrate J Food Sci 51 (3) 564-67

Ballance GM WOS Meredith 1976 Purif ication and partial characterization of an endo- β -13-glucanase from green malt J Inst Brew 8264-67

Bamforth CW and AHP Barclay 1993 Malting technology and the uses of malt In Barley Chemistry and Technology (eds AW MacGregor and RS Bhatty) by Am Assoc Cereal Chem St Paul USA pp 297-354

Bansema C 2000 Development of a barley P-glucan beverage with and without whey protein Isolate MSc thesis Edmonton Alberta Canada

Basman A and HK Ksel 1999 Properties and composit ion of Turkish f lat bread (bazlama) supplemented with barley f lour and wheat bran Cereal Chem 76506ndash511

Beer MU E Arrigoni and R Amado 1995 Effect of oat gum on blood cholesterol levels in healthy young men Europ J Clin Nutri 49517ndash522

140

Beer MU PJ Wood J Weisz N Fi l l ion 1997 Effect of cooking and storage on the amount and molecular weight of (1rarr3) (1rarr4) - β -D-glucan extracted from oat products by an in vitro digestion system Cereal Chem 74 705-709

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 A Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bender DA and AE Bender 1999 Bendersrsquo Dictionary of Nutrit ion and Food Technology 7 t h ed Woodhead Publishing Abington

Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

Berglund PT CE Fastnaught ET Holm 1992 Food uses of waxy hull- less barley Cereal Foods World 37707ndash714

Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

Bhatty RS 1992 Total and extractable β -glucan contents of oats and their relationship to viscosity J Cer Sci 15185-192

Bhatty RS 1995 Laboratory and pilot plant extraction and purif ication of b-glucans from hull- less barley and oat bran J Cer Sci 22163ndash170

Bhatty RS 1996 Production of food malt from hull- less barley Cereal Chem 73(1) 75-80

Bhatty RS AW MacGregor and BG Rossnagel 1991 Total and acid-soluble β -glucan content of hulless barley and its relationship to acid-extract viscosity Cereal Chem 68221-227

Bhatty RS1986 Physiochemical and Functional (Breadmaking) Properties of Hull- less Barley Fractions Cereal Chem 6331-35

141

Bibek R 2001 Fundamental Food Microbiology 2nd edn The CRC press Ltd Washington DC pp 56-90

Bingham SA NE Day R Luben P Ferrari N Sl imani T Norat F Lavel E Kesse A Nieters H Boeing A Tjoslashnneland K Overvad C Martinez M Dorrensoro CA Gonzalez TJ Key A Trichopoulou A Naska P Vineis R Tumino V Krogh HB Bueno-de-Mesquita PHM Peeters G Berglung G Hallmans E Lund G Skele R Kaaks and E Riboli 2003 Dietary f ibre in food and protection against colorectal cancer in the European Prospective Investigation into Cancer and Nutrit ion (EPIC) an observational study Lancet 3611496-501

Bioumlrklund M A van Rees RP Mensink and G Oumlnning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucans from oats and barley a randomised dose-controlled tr ial Eur J Clin Nutri 591272-1281

Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

Bjorck I AC Eliasson A Drews M Gudmundsson and R Karlsson 1990 some nutrit ional properties of starch and dietary f iber in barley genotypes containing different levels of amylose Cereal Chem 67 327

BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

Brand J S Colagiuri S Crossman A Allen D Roberts and S Truswell 1991 Low-glycemic index foods improve long term glycemic control in NIDDM Diabetes Care 14 95ndash101

142

Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

Brennan CS and LJ Cleary 2005 The potential use of cereal (1314)- β -D-glucans as functional food ingredients J Cer Sci 421ndash13

Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

Brown L B Rosner W Willet and FM Sacks 1999 Cholesterol lowering effects of dietary f iber a meta analysis Am J Clin Nutri 69 (1) 30 42

Brunswick P DJ Manner and J K Stark 1987 Development of β -D-glucanases during germination of barley and the effect of ki lning on individual isoenzymes J Inst Brew 93181-186

Bryan D J Robert AT Wilson T Carlson S Frazer GH Zheng 2003 β -Glucan Fractions from Barley and Oats Are Similarly Antiatherogenic in Hypercholesterolemic Syrian Golden Hamsters The American Society for Nutrit ional Sciences J Nutri Metabolism 133468-475

Buliga GS DA Brant and GB Fincher 1986 The sequence statist ics and solution configration of barley (1rarr3) (1rarr4) - β -D-glucan Carbohydr Res 57139-156

Burkus Z 1996 Barley P-Glucan Extraction Functional Properties and Interactions with Food Components MSc thesis Edmonton AlbertaCanda

Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloid F Glicksman M (Ed) p 49-93 CRC Press Inc Boca Raton

Burkus Z 1996 Barley β -glucan Extraction Functional properties and interaction with food components MSc Thesis Dept of Agricultural Food and Nutrit ional Science Univ of Alberta Edmonton Canada

143

Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

Burkus Z and F Temell i 1999 Glucan concentrate J Food Sci 64198-201 Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloidr Glicksman M (Ed) p 49-93 CRC Press hc Boca Raton FL

Burkus Z and F Temell i 2005 Rheological properties of barley β -glucan Carbohydr Polym 59 459ndash465

Burkus Z F Temell i 1999 Gelation of barley β -glucan - concentrate J Food Sci 64198-201

Calix FD and N Bardrie 2004 Consumer acceptance and physicochemical quality of processed red sorrelroselle (Hibiscus sabdar i f fa L) sauces from enzymatic extracted calyces 4 141-148

Carpita NC 1996 Structure and biogenesis of cel l walls of grasses Annual Rev Plant Physiol Plat Molecular Biol 47445-476

Carr J M S Glatter J L Jeraci and B A Lewis 1990 Enzymes Determination of Beta-Glucan in Cereal-Based Food Products Cereal Chem 67226-229

Casterl ine J L CJ Oles and Y Ku 1997 In vitro fermentation of various food f iber reactions J Agric Food Chem 452463ndash2467

Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

Chowdhury MGF MN Islam MS Is lam T Is lam and MS Hossain 2008 Study on Preparation and Shelf-Life of Mixed Juice Based on Wood Apple and Papaya J Soil Nature 2(3) 50-60

Chung OK and Y Pomeranz 1985 Amino acids in cereal proteins and protein fractions Ch 5 in Digesfibi l i~ and

144

Amino Acid Availabil i ty in Cereals andOilseeds J W Finley and DT Hopkins (Eds) pp 169-232 AACC St Paul MN

Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

Crandall PG CS Chen and KC Davis 1987 Preparation and storage of 72 brix orange juice concentration J Food Sci 52 (3) 381

Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

Daw ZY YSA El-Gizaw and AMB Said 1994 Microbiological evaluation of some local juices and drinks Chemie Mikrobiologie Technologie der Lebensmittel 168ndash15

Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

Dawkins NL I A Nnanna 1993 Studies on oat gum [(1rarr31rarr4)- β-D-glucan] Composit ion molecular weight est imation and rheological properties Food Hydrocol 9 1-7

Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

Dohnalek MH 2004 The role of f ibre in cl inical nutrit ion In Van der Kamp JW Asp NG Miller J J Schaafsma G (Ed) Dietary f ibre bioactive carbohydrates for food and feed Wageningen Academic Publishers Wageningen pp 271294

145

Dongowski G M Huth E Gebhardt and W Flamme 2002 Dietary f iber-rich barley products beneficial ly affect the intestinal tract of rats J Nutri 132(12) 3704-14

Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

Falade OS OR Sowunmi A Oladipo A Tobosun and SRA Adewusi 2003 The level of organic acids in some Nigerian fruit and their effect on mineral availabil i ty in composite diet Pak J Nutri 2(2) 82-83

Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

FDA 1996 Food labeling Health claims oats and coronary heart disease Federal Register 61 (3) January 4

Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

Frazier WC and EM Foster 1958 Laboratory Manual for Food Microbiology Burgess Pub Co Minneapolis Minnisota USA

Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

146

Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

Giese J H 1992 Hitt ing the spot Beverages and beverage technology Food Technol 4670-72 74-75 78-80

Godara RK and OP Pareek 1985 Effect of temperature in storage of ready to serve date juice beverages indian j agric Sci 55 (5) 347-349 (FSTA 18 (4) 78 1986)

Gonzalez ER and S Leeson 2000 An investigation on the preservation of kununndashzaki an African fermented cereal based food drink Acta Alimentaria 29 385ndash92

GOP 2008 Government of Pakistan Finance Division Economic Advisor s Wing Islamabad Pakistan

Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

Hallfr isch J DJ Schofield KM Behall 2003 Physiological responses of men and women to barley and oat extracts (NutrimX) I I Comparison of glucose and insulin responses Cereal Chem 8080ndash83

Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

147

Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

Lawless HT and H heymann Sensory evaluation of food Principles and Practices Gaithersburg MD Aspen Publishers ISSN 1572-0330) Oorspr uitg New York [etc ] Chapman amp Hall 1998

Hashimoto S MD Shogren Y Pomeranz 1987 Cereal Pentosans Their est imation and signif icance I Pentosans in wheat and milled wheat products Cereal Chem 64(1) 30-34

Hassan SA 1976 Effect of storage on physico-chemical characterist ics of carbonated orange juice Msc thesis Food Tech Deptt WPAU Lyallpur

Hatcher WSJ R J L Weihe DF Split tstoesser EC Hil l and ME Parish 1992 Fruit Beverages In Compendium of methods for the microbiological examination of foods Vanderzant C Split tstoesser DF (eds) American Public Health Association Washington DC

Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

Hil l M J and FR Path 1998 Cereals dietary f iber and cancer Nutri Res 18563ndash659

Hil l iam M 2000 Functional foodndashndashHow big is the market The World of Food Ingredients 12 50ndash2

Holsinger V H LP Posati and ED DeVilbiss 1974 Whey beverages a review J Dairy Sci 57(7) 849ndash859

Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

148

47ndash82) St Paul MN USA American Association of Cereal Chemists

Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

Jadhav SJ S E Lutz VM Ghorpade and DK Salunkhe 1998 Barley chemistry and value-added processing Crit ical Rev Food Sci 3823ndash171

Jal i l i T REC Wildman DM Medeiros 2000 Nutraceutical roles of dietary f iber J Nutraceutical functional and Medi foods 2 19-34

Jansen MC HB Bueno-de-Mesquita R Buzina F Fidanza A Menotti H Blackburn AM Nissinen FJ Kok D Kromhout 1999 Dietary f iber and plant foods in relation to colorectal cancer mortal i ty The Seven Countries Study Inter J Canc 81 174-179

Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high

List of Figures

Fig Title Page

31 Preparation of β -glucan beverage 47 41 Percent decrease in the serum total cholesterol level of subjects fed

on different beverages 111

42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of healthy volunteers

112

43 Percent decrease in the serum triglycerides level of subjects fed on different beverages

115

44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy volunteers

116

45 Percent decrease in the serum LDL level of subjects fed on different beverages

119

46 Effect of β-glucan beverage on LDL (mgdl) content of healthy volunteers

120

47 Percent increase in the serum HDL level of subjects fed on different beverages

123

48 Effect of β-glucan beverage on HDL (mgdl) content of healthy volunteers

124

49 Effect of β-glucan beverage on blood glucose (mgdl) content of healthy volunteers

128

List of Appendices

Appendix Title Page

I Composit ion of functional beverage 162

II 9 Point Hedonic Scale 163

III Food frequency questionnaire 164

IV Demographic information performa (subjects) 165

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

LITERATURE CITED

AACC 2000 Approved Methods of American Association of Cereal Chemists The American Association of Cereal Chemists Inc St Paul Minnesota USA

Aastrup S 1979a The effect of rain on β -glucan content in barley grains Carlsberg esearch Communications 44381-393

Aditya K T Yokota S Suzuki and H Etoh 2008 Sub crit ical Water Extraction of Barley to Produce a Functional Drink

Biosci Biotechnol Biochem 72(1)236-239

AERI 1896 The Agricultural Economics Research Institute Balance Sheet for Food Commodities Finland 1985 The Insti tute Helsinki

Akubor PI 2003 Influence of storage on the physicochemical microbiological and sensory properties of heat and chemically treated melon-banana beverage Plant Foods for Human Nutri 58 1ndash10

Alessandra DC P Antonio V Vincenzo A Mario 2004 Changes of f lavonoids vitamin C and antioxidant capacity in minimally processed citrus segments and juices during storage Food Chem 84 99-105

Aman P H Graham AC Til ly 1989 Content and solubil i ty of mixed-l inked (1-3) (1-4)- β -D-glucan in barley and oats during kernel development and storage J Cereal Sci 1045-50

Anderson J W 1980 Dietary f iber and diabetes in Medical Aspects of Dietry Fiber G A spil ler and R M Key eds Plenum Medical Book Company New York

Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

Anderson J W DB Spencer CC Hamilton SF Smith and J Tietyen CA Bryant P Oeltgen 1990 Oat-bran cereal lowers serum total and LDL cholesterol in hypercholesterolemic men Am J Clin Nutri 52 495-499

139

Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

Annoni G BM Botasso D Ciaci MF Donato and A Tripodi 1982 Liquid tr iglycerides (GPO-PAP) Medi Diagnostic I taly Lab J Res Lab Med 9 115-116

AOAC 2000 Official Methods of Analysis The Association of the Official Analytical Chemists 20 t h Ed Arlington USA

Arndt EA 2006 Whole-grain barley for todays health and wellness needs ConAgra Foods Inc Omaha NE 51(1) 20-22

Assmann G 1979 HDL-cholesterol precipitant Randox Labs Ltd CrumLin Co Antrim N Ireland Internist 20559-567

Babsky NE J L Toribio and J E Lozano 1986 Influence of storage on the composit ion of clarif ied apple juice concentrate J Food Sci 51 (3) 564-67

Ballance GM WOS Meredith 1976 Purif ication and partial characterization of an endo- β -13-glucanase from green malt J Inst Brew 8264-67

Bamforth CW and AHP Barclay 1993 Malting technology and the uses of malt In Barley Chemistry and Technology (eds AW MacGregor and RS Bhatty) by Am Assoc Cereal Chem St Paul USA pp 297-354

Bansema C 2000 Development of a barley P-glucan beverage with and without whey protein Isolate MSc thesis Edmonton Alberta Canada

Basman A and HK Ksel 1999 Properties and composit ion of Turkish f lat bread (bazlama) supplemented with barley f lour and wheat bran Cereal Chem 76506ndash511

Beer MU E Arrigoni and R Amado 1995 Effect of oat gum on blood cholesterol levels in healthy young men Europ J Clin Nutri 49517ndash522

140

Beer MU PJ Wood J Weisz N Fi l l ion 1997 Effect of cooking and storage on the amount and molecular weight of (1rarr3) (1rarr4) - β -D-glucan extracted from oat products by an in vitro digestion system Cereal Chem 74 705-709

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 A Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

Bender DA and AE Bender 1999 Bendersrsquo Dictionary of Nutrit ion and Food Technology 7 t h ed Woodhead Publishing Abington

Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

Berglund PT CE Fastnaught ET Holm 1992 Food uses of waxy hull- less barley Cereal Foods World 37707ndash714

Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

Bhatty RS 1992 Total and extractable β -glucan contents of oats and their relationship to viscosity J Cer Sci 15185-192

Bhatty RS 1995 Laboratory and pilot plant extraction and purif ication of b-glucans from hull- less barley and oat bran J Cer Sci 22163ndash170

Bhatty RS 1996 Production of food malt from hull- less barley Cereal Chem 73(1) 75-80

Bhatty RS AW MacGregor and BG Rossnagel 1991 Total and acid-soluble β -glucan content of hulless barley and its relationship to acid-extract viscosity Cereal Chem 68221-227

Bhatty RS1986 Physiochemical and Functional (Breadmaking) Properties of Hull- less Barley Fractions Cereal Chem 6331-35

141

Bibek R 2001 Fundamental Food Microbiology 2nd edn The CRC press Ltd Washington DC pp 56-90

Bingham SA NE Day R Luben P Ferrari N Sl imani T Norat F Lavel E Kesse A Nieters H Boeing A Tjoslashnneland K Overvad C Martinez M Dorrensoro CA Gonzalez TJ Key A Trichopoulou A Naska P Vineis R Tumino V Krogh HB Bueno-de-Mesquita PHM Peeters G Berglung G Hallmans E Lund G Skele R Kaaks and E Riboli 2003 Dietary f ibre in food and protection against colorectal cancer in the European Prospective Investigation into Cancer and Nutrit ion (EPIC) an observational study Lancet 3611496-501

Bioumlrklund M A van Rees RP Mensink and G Oumlnning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucans from oats and barley a randomised dose-controlled tr ial Eur J Clin Nutri 591272-1281

Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

Bjorck I AC Eliasson A Drews M Gudmundsson and R Karlsson 1990 some nutrit ional properties of starch and dietary f iber in barley genotypes containing different levels of amylose Cereal Chem 67 327

BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

Brand J S Colagiuri S Crossman A Allen D Roberts and S Truswell 1991 Low-glycemic index foods improve long term glycemic control in NIDDM Diabetes Care 14 95ndash101

142

Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

Brennan CS and LJ Cleary 2005 The potential use of cereal (1314)- β -D-glucans as functional food ingredients J Cer Sci 421ndash13

Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

Brown L B Rosner W Willet and FM Sacks 1999 Cholesterol lowering effects of dietary f iber a meta analysis Am J Clin Nutri 69 (1) 30 42

Brunswick P DJ Manner and J K Stark 1987 Development of β -D-glucanases during germination of barley and the effect of ki lning on individual isoenzymes J Inst Brew 93181-186

Bryan D J Robert AT Wilson T Carlson S Frazer GH Zheng 2003 β -Glucan Fractions from Barley and Oats Are Similarly Antiatherogenic in Hypercholesterolemic Syrian Golden Hamsters The American Society for Nutrit ional Sciences J Nutri Metabolism 133468-475

Buliga GS DA Brant and GB Fincher 1986 The sequence statist ics and solution configration of barley (1rarr3) (1rarr4) - β -D-glucan Carbohydr Res 57139-156

Burkus Z 1996 Barley P-Glucan Extraction Functional Properties and Interactions with Food Components MSc thesis Edmonton AlbertaCanda

Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloid F Glicksman M (Ed) p 49-93 CRC Press Inc Boca Raton

Burkus Z 1996 Barley β -glucan Extraction Functional properties and interaction with food components MSc Thesis Dept of Agricultural Food and Nutrit ional Science Univ of Alberta Edmonton Canada

143

Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

Burkus Z and F Temell i 1999 Glucan concentrate J Food Sci 64198-201 Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloidr Glicksman M (Ed) p 49-93 CRC Press hc Boca Raton FL

Burkus Z and F Temell i 2005 Rheological properties of barley β -glucan Carbohydr Polym 59 459ndash465

Burkus Z F Temell i 1999 Gelation of barley β -glucan - concentrate J Food Sci 64198-201

Calix FD and N Bardrie 2004 Consumer acceptance and physicochemical quality of processed red sorrelroselle (Hibiscus sabdar i f fa L) sauces from enzymatic extracted calyces 4 141-148

Carpita NC 1996 Structure and biogenesis of cel l walls of grasses Annual Rev Plant Physiol Plat Molecular Biol 47445-476

Carr J M S Glatter J L Jeraci and B A Lewis 1990 Enzymes Determination of Beta-Glucan in Cereal-Based Food Products Cereal Chem 67226-229

Casterl ine J L CJ Oles and Y Ku 1997 In vitro fermentation of various food f iber reactions J Agric Food Chem 452463ndash2467

Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

Chowdhury MGF MN Islam MS Is lam T Is lam and MS Hossain 2008 Study on Preparation and Shelf-Life of Mixed Juice Based on Wood Apple and Papaya J Soil Nature 2(3) 50-60

Chung OK and Y Pomeranz 1985 Amino acids in cereal proteins and protein fractions Ch 5 in Digesfibi l i~ and

144

Amino Acid Availabil i ty in Cereals andOilseeds J W Finley and DT Hopkins (Eds) pp 169-232 AACC St Paul MN

Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

Crandall PG CS Chen and KC Davis 1987 Preparation and storage of 72 brix orange juice concentration J Food Sci 52 (3) 381

Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

Daw ZY YSA El-Gizaw and AMB Said 1994 Microbiological evaluation of some local juices and drinks Chemie Mikrobiologie Technologie der Lebensmittel 168ndash15

Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

Dawkins NL I A Nnanna 1993 Studies on oat gum [(1rarr31rarr4)- β-D-glucan] Composit ion molecular weight est imation and rheological properties Food Hydrocol 9 1-7

Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

Dohnalek MH 2004 The role of f ibre in cl inical nutrit ion In Van der Kamp JW Asp NG Miller J J Schaafsma G (Ed) Dietary f ibre bioactive carbohydrates for food and feed Wageningen Academic Publishers Wageningen pp 271294

145

Dongowski G M Huth E Gebhardt and W Flamme 2002 Dietary f iber-rich barley products beneficial ly affect the intestinal tract of rats J Nutri 132(12) 3704-14

Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

Falade OS OR Sowunmi A Oladipo A Tobosun and SRA Adewusi 2003 The level of organic acids in some Nigerian fruit and their effect on mineral availabil i ty in composite diet Pak J Nutri 2(2) 82-83

Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

FDA 1996 Food labeling Health claims oats and coronary heart disease Federal Register 61 (3) January 4

Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

Frazier WC and EM Foster 1958 Laboratory Manual for Food Microbiology Burgess Pub Co Minneapolis Minnisota USA

Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

146

Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

Giese J H 1992 Hitt ing the spot Beverages and beverage technology Food Technol 4670-72 74-75 78-80

Godara RK and OP Pareek 1985 Effect of temperature in storage of ready to serve date juice beverages indian j agric Sci 55 (5) 347-349 (FSTA 18 (4) 78 1986)

Gonzalez ER and S Leeson 2000 An investigation on the preservation of kununndashzaki an African fermented cereal based food drink Acta Alimentaria 29 385ndash92

GOP 2008 Government of Pakistan Finance Division Economic Advisor s Wing Islamabad Pakistan

Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

Hallfr isch J DJ Schofield KM Behall 2003 Physiological responses of men and women to barley and oat extracts (NutrimX) I I Comparison of glucose and insulin responses Cereal Chem 8080ndash83

Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

147

Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

Lawless HT and H heymann Sensory evaluation of food Principles and Practices Gaithersburg MD Aspen Publishers ISSN 1572-0330) Oorspr uitg New York [etc ] Chapman amp Hall 1998

Hashimoto S MD Shogren Y Pomeranz 1987 Cereal Pentosans Their est imation and signif icance I Pentosans in wheat and milled wheat products Cereal Chem 64(1) 30-34

Hassan SA 1976 Effect of storage on physico-chemical characterist ics of carbonated orange juice Msc thesis Food Tech Deptt WPAU Lyallpur

Hatcher WSJ R J L Weihe DF Split tstoesser EC Hil l and ME Parish 1992 Fruit Beverages In Compendium of methods for the microbiological examination of foods Vanderzant C Split tstoesser DF (eds) American Public Health Association Washington DC

Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

Hil l M J and FR Path 1998 Cereals dietary f iber and cancer Nutri Res 18563ndash659

Hil l iam M 2000 Functional foodndashndashHow big is the market The World of Food Ingredients 12 50ndash2

Holsinger V H LP Posati and ED DeVilbiss 1974 Whey beverages a review J Dairy Sci 57(7) 849ndash859

Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

148

47ndash82) St Paul MN USA American Association of Cereal Chemists

Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

Jadhav SJ S E Lutz VM Ghorpade and DK Salunkhe 1998 Barley chemistry and value-added processing Crit ical Rev Food Sci 3823ndash171

Jal i l i T REC Wildman DM Medeiros 2000 Nutraceutical roles of dietary f iber J Nutraceutical functional and Medi foods 2 19-34

Jansen MC HB Bueno-de-Mesquita R Buzina F Fidanza A Menotti H Blackburn AM Nissinen FJ Kok D Kromhout 1999 Dietary f iber and plant foods in relation to colorectal cancer mortal i ty The Seven Countries Study Inter J Canc 81 174-179

Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high

List of Appendices

Appendix Title Page

I Composit ion of functional beverage 162

II 9 Point Hedonic Scale 163

III Food frequency questionnaire 164

IV Demographic information performa (subjects) 165

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

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Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

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Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

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Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

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Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

Berglund PT CE Fastnaught ET Holm 1992 Food uses of waxy hull- less barley Cereal Foods World 37707ndash714

Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

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Bhatty RS 1995 Laboratory and pilot plant extraction and purif ication of b-glucans from hull- less barley and oat bran J Cer Sci 22163ndash170

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Bingham SA NE Day R Luben P Ferrari N Sl imani T Norat F Lavel E Kesse A Nieters H Boeing A Tjoslashnneland K Overvad C Martinez M Dorrensoro CA Gonzalez TJ Key A Trichopoulou A Naska P Vineis R Tumino V Krogh HB Bueno-de-Mesquita PHM Peeters G Berglung G Hallmans E Lund G Skele R Kaaks and E Riboli 2003 Dietary f ibre in food and protection against colorectal cancer in the European Prospective Investigation into Cancer and Nutrit ion (EPIC) an observational study Lancet 3611496-501

Bioumlrklund M A van Rees RP Mensink and G Oumlnning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucans from oats and barley a randomised dose-controlled tr ial Eur J Clin Nutri 591272-1281

Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

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BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

Brand J S Colagiuri S Crossman A Allen D Roberts and S Truswell 1991 Low-glycemic index foods improve long term glycemic control in NIDDM Diabetes Care 14 95ndash101

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Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

Brennan CS and LJ Cleary 2005 The potential use of cereal (1314)- β -D-glucans as functional food ingredients J Cer Sci 421ndash13

Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

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Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

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Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

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Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

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Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

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Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

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Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

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Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

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Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

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Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

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Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

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Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

Giese J H 1992 Hitt ing the spot Beverages and beverage technology Food Technol 4670-72 74-75 78-80

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Gonzalez ER and S Leeson 2000 An investigation on the preservation of kununndashzaki an African fermented cereal based food drink Acta Alimentaria 29 385ndash92

GOP 2008 Government of Pakistan Finance Division Economic Advisor s Wing Islamabad Pakistan

Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

Hallfr isch J DJ Schofield KM Behall 2003 Physiological responses of men and women to barley and oat extracts (NutrimX) I I Comparison of glucose and insulin responses Cereal Chem 8080ndash83

Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

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Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

Lawless HT and H heymann Sensory evaluation of food Principles and Practices Gaithersburg MD Aspen Publishers ISSN 1572-0330) Oorspr uitg New York [etc ] Chapman amp Hall 1998

Hashimoto S MD Shogren Y Pomeranz 1987 Cereal Pentosans Their est imation and signif icance I Pentosans in wheat and milled wheat products Cereal Chem 64(1) 30-34

Hassan SA 1976 Effect of storage on physico-chemical characterist ics of carbonated orange juice Msc thesis Food Tech Deptt WPAU Lyallpur

Hatcher WSJ R J L Weihe DF Split tstoesser EC Hil l and ME Parish 1992 Fruit Beverages In Compendium of methods for the microbiological examination of foods Vanderzant C Split tstoesser DF (eds) American Public Health Association Washington DC

Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

Hil l M J and FR Path 1998 Cereals dietary f iber and cancer Nutri Res 18563ndash659

Hil l iam M 2000 Functional foodndashndashHow big is the market The World of Food Ingredients 12 50ndash2

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Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

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47ndash82) St Paul MN USA American Association of Cereal Chemists

Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

Jadhav SJ S E Lutz VM Ghorpade and DK Salunkhe 1998 Barley chemistry and value-added processing Crit ical Rev Food Sci 3823ndash171

Jal i l i T REC Wildman DM Medeiros 2000 Nutraceutical roles of dietary f iber J Nutraceutical functional and Medi foods 2 19-34

Jansen MC HB Bueno-de-Mesquita R Buzina F Fidanza A Menotti H Blackburn AM Nissinen FJ Kok D Kromhout 1999 Dietary f iber and plant foods in relation to colorectal cancer mortal i ty The Seven Countries Study Inter J Canc 81 174-179

Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high

ABSTRACT

The research project was carried out to explore the health

benefi ts of barley β -glucan in beverage Beverages were prepared

with different levels of β -glucan and then analyzed for various

quali ty attr ibutes during storage The L a and b value for color of

beverages increased signif icantly by increasing the level of β -glucan

The highest viscosity (2175 mPa-s) and total soluble sol ids

(1042ordmbrix) were found in T6beverage containing 1 β -glucanThe

pH decreased signif icantly in al l beverages throughout the storage

period Total acidity and ascorbic acid varied signif icantly as a

function of storage The reducing sugars increased from 372 to 4 31

from 0 to 90 days of storage respectively The total plate count of

beverages decreased from 129 times 10 4 to 1 17 times 10 4 at the end of the

storage The scores assigned to al l the sensory parameters of

beverages affected signif icantly with the variat ion in the levels of β -

glucan and decreased signif icantly during storage intervals The

treatments T2 T3 and T4 got containing 0 2 0 4 and 06 β -glucan

got highest scores for sensory evaluation Total cholesterol glucose

LDL-C and tr iglyceride contents in serum of adult humans fed on

beverages decreased signif icantly whereas concentrat ion of HDL

improved due to incorporation of β -glucan in beverages The

beverage with 0 6 β -glucan contributed to reduce the serum

glucose of human subjects by 1018 cholesterol by 8 26

tr iglycerides by 1099 and LDL by 1082 The present study

suggests that β -glucan is a funct ional ingredient and can be used to

prevent cardiovascular diseases and also to control diabetes

1

CHAPTER-1

INTRODUCTION

Cereals are considered one of the most important economic

and food commodities in the world The cereals grains are

harvested over 1 bi l l ion tones annually The barley (Hordeum

vulgare L ) accounts for 12 of the worlds total cereal production

and occupies fourth posit ion with respect to grain production

after wheat r ice and corn (Jadhav et a l 1998) The barley grain

was produced 13747 mil l ion metric tones in the world during the

crop year 2006-2007(FAS 2008) The leading barley producing

countries in the world are EU countries (5165 mil l ion tones)

fol lowed by the Russian Federat ion (2501 mil l ion tones) and

Canada (1317 mil l ion tones) (Brennan and Cleary 2005) In

Pakistan production of barley grain was 98000 tones harvested

from an area of 92000 hectares during the crop year 2007-08

(GOP 2007-08) In world approximately 81 of annual barley

production is used for feed 9 for seed 8 for malt and alcohol

production and only 2 is used for human consumption (AERI

1986) Like other countries this crop is also mainly goes for

feeding the animals and its human consumption is very l imited in

Pakistan The variet ies such as Jau-83 Jau-87 Haider-93 and some

promising hulless l ines of barley developed are being cult ivated

commercial ly in Pakistan

Barley is gett ing renewed interest as an ingredient in the

production of functional foods due to i ts higher content of

bioactive compounds Barley possesses high amount of dietary

2

f iber (DF) with high proportion of soluble viscous components

offering more suitabil i ty among cereal grains in the human diet

(Bjorck et a l 1990) The barley in the world is used mainly as an

animals feed in the form of barley meal and as grain for malting

and brewing for manufacturing of beer and whisky The research

has been focussed mainly on assessing the role of endospermic

components in relation to malting potential of barley grain

(Molina-Cano et a l 2002) However the barley grain has been

relatively under-uti l ized with respect to i ts potential use as a

human food The potential use of β -glucan extracted from barley

and other cereal grains as a functional ingredient in different

foods has received more attention in the recent years (Malkki

2004) There are some new waxy hulless barley variet ies l ike

Prowashonupana have also been developed which possess unique

macronutrient composit ion with higher content of f iber and

protein and lower amount of starch as compared to other common

cereal grains The barley can potential ly be used to develop and

formulate products with improved health benefits and a variety of

health c laims This particular barley grains can be used to

enhance the f lavor texture appearance and nutrit ional

composit ion for a variety of food product applications including

hot cereals cookies crackers breads tort i l las granola bars fruit-

f i l led cereal bars extruded snacks and pastas The functional

f lexibil i ty of barley al lows it to be used in foods that span across

meal occasions including muffins and ready-to eat cereals for

breakfast soup vegetarian patt ies and pizza crackers and

extruded chips for snacks and cookies and toppings for dessert

and development of different beverages ( Arndt 2006)

3

The barley contains substantial ly higher amounts of

functional ingredient i e β -glucan but oat and some fungi and

moulds also possess good amount of β -glucans The use of β -

glucan extracted from barley as a human food due to i ts posit ive

role in human health has received a growing attention The cel l

wall of barley and oat contains β -glucan a non starch

polysaccharide composed of β - (1-4)- l inked glucose units

separated every two to three units by a single β - (1-3)ndashl inked

glucose and referred to as a mixed l inkage β -glucan (Carpita

1996)

In human diet the health promoting properties of β-glucan

have been demonstrated High-serum cholesterol one of the

important r isk factor for coronary heart disease (Anderson 1986)

is reduced by the intake of β -glucan which wil l ult imately the

risk of cardiovascular diseases The soluble dietary f iber

component may assist in regulation of blood glucose and lowering

of serum cholesterol (Anderson 1980) The β -glucan a soluble

f iber extracted from oat or consumed as oat porridge reduced

postprandial blood glucose (Wood et at 1990) β -glucan delays

glucose absorption which regulates the level of blood glucose

(Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property of β -glucan

may be useful in the formulation of food products targeting

management of diabetes

In recent years human health has received an unprecedented

important status The interests in nutrit ion f i tness and beauty

have main concerns over diet and human health in todayrsquos l iving

style The foods which should provide additional physiological

4

benefits such as preventing or delaying onset of chronic diseases

besides meeting basic nutrit ional requirements are known as

functional foods (Nicoli et a l 1999) Functional foods including

functional beverages are important for their role in health

promotion and disease prevention The functional foods are not

intended only to satisfy hunger but also provid necessary

nutrients to human for prevention of nutrit ion-related diseases

(Menrad et a l 2000) The growing interest in new functional

foods with special characterist ics and health benefits has led to

the development of new functional beverages The global market

of functional food has been estimated to be at least 33 bi l l ion US$

(Hil l iam 2000)

The functional beverages can play an important role in

health promotion and disease prevention They provide means to

reduce the increasing burden on the health care system by a

continuous preventive mechanism (Shahidi 2004) The functional

beverages not only provide taste and refreshment satisfaction but

can also provide necessary nutrients to prevent nutrit ion-related

diseases (Menrad et a l 2000) Beverages are considered to be an

excellent medium for the supplementation of nutraceutical

components for enrichment (Kuhn 1998) such as soluble f iber or

herbal extract (Swientek 1998)

The functional beverage may enrich the diet and improve

health of human because of i t ease of consumption along with a

usual meal Barley β -glucan assume to be well suited for such an

functional application being capable of imparting a smooth

mouth feel to beverage products and providing an excellent

source of soluble dietary f iber A barley β -glucan gum with

5

similar functional properties could potential ly serve as an

alternative to tradit ional beverage thickeners such as alginates

pectin xanthan and carboxymethylcel lulose (Giese 1992)

Barley tea is a common drink in Japan especial ly during the

summer This non-caffeinated non-tannin drink is valued for i ts

high percentage of β - glucan (polysaccharides) and the presence

of antioxidant compounds (Etoh et a l 2004 Tsunagi et a l 2003)

The use of β -glucan due to i ts good viscosity forming properties

offer potential alternatives as thickening agents in different food

applications e g ice creams sauces and salad dressings (Wood

1986) The uti l ization of barley β -glucan as an ingredient in the

production of a functional beverage has not been fully exploited

so far

The nutrit ional and functional benefits of β -glucan including

thickening stabil izing emulsif ication and gelation revealed that

β -glucan from barley can be used for the preparation of functional

beverage Therefore this study was planned to extract the β -

glucan from Pakistani barley variety (Haider-93) and its

uti l ization for the development of functional beverage Therefore

the mandate of the present study was as under

bull To develop a suitable formulation and processing procedure for a functional beverage with incorporation of barley β- glucan

bull To evaluate quality parameters and acceptabil i ty of functional beverage

bull To examine the shelf stabil i ty of β -glucan beverage using instrumental techniques

bull To evaluate the effect of β -glucan beverage on the glucose level and l ipid profi le of human volunteers

6

CHAPTER-2

REVIEW

OF

LITERATURE

Cereal β -glucan is a soluble dietary f iber and offers

potential for food products The beverages are one of the best

media for incorporation of β -glucan The characterist ic properties

desired in the beverage such as color f lavor and mouth feel make

the barley β -glucan an ideal grain over other cereals such as

sorghum and wheat (Bamforth and Barclay 1993) I t also exhibits

some health benef its such as lowering of blood glucose level and

prevention of cardiovascular diseases By manipulating the β -

glucan and protein contents of barley numerous types of malt

(beer) and other beverages are l ikely to satisfy various human

tastes (Munk 1981)

The l i terature pertaining to different aspects of the present

study is reviewed under fol lowing headings

2 1 Barley History composit ion and types

22 Role of dietary f iber

23 β -glucan Sources and occurrence

2 4 β -glucan extraction

7

25 Health benefits of β -glucan

26 Functional properties of β -glucan

27 Uti l ization of β -glucan in food products

28 Physico-chemical characterist ics of beverages

21 Barley History composition and types

The cereals are defined as edible seeds of the grass family

Gramineae (Bender and Bender 1999) The cereals are cult ivated

for their nutrit ious edible seeds often referred as grains and

used as staple food for the human consumption and l ivestock feed

since the early civi l ization (BNF 1994) Cereal grains contribute

signif icant amounts of energy protein and micronutrients to the

human diet and contain a large number of biologically active

substances including antioxidants dietary f iber phytoestrogens

and l ignans (Hil l and Path 1998)

Barley (Hordeum vulgare L ) competes with wheat regarding

the most ancient cereal crop I t referred as the original ancient

cereal grains consumed around the world throughout the history

Barley has been recorded as being cult ivated along the Nile River

thousands of years ago dating back to Egyptian t imes (Wendorf et

a l 1979) Barley is an old crop and its cult ivation mentioned in

the Bible Due to i ts cold drought alkali and salt tolerance i t is

grown at 70degN lati tude in Norway as well as in regions close to

the equator at high alt i tudes (Poehlman 1985) With respect to

world cereal grain production barley ranks fourth fol lowed by

wheat r ice and corn (Nilan and Ullrich 1993) Barley is a major

crop for malt ing brewing and for food production industries in

8

the developed countries and it is uti l ize as fodder crop in the less

developed and developing countries (Kent and Evers 1994)

Barley is a typical cereal grain composed primarily of starch

protein f iber l ipids and minerals The typical composit ion of

barley is outl ined in Table 21 (MacGregor and Fincher 1993)

Barley is a source of protein typically contains 10-12 in the

whole grain containing more of the essential amino acids

particularly lysine which is the f irst l imiting amino acid in the

wheat (Chung and Pomeranz 1985) Barley proteins can be

grouped as storage and non-storage proteins Storage proteins

include the prolamins (hordeins) and globulins as defined by

Osborne protein classif ication (Shewry 1993) Being high

molecular weight water soluble polymers they have unique

properties with both nutri t ional and technological s ignif icance

They are not digested by mono gastric animal which is one reason

for the low use of barley as poultry feed (Wood 1984) I t has

recently been rediscovered as a nutrit ious food grain for the

human diet and is expected to see some increase in food

applications in the near future The starch portion of the grain is a

good source of digestible carbohydrate necessary for energy

(MacGregor and Fincher 1993)

There are generally two types of barley hulled and hull- less

barley Hull- less barley contains more protein starch and β -

glucan than hulled barley I t is a good source of f iber in general

and of soluble f iber such as β -glucan in particular (Bhatty 1999)

Most of the barley used in the world today is covered (Hulled) as

covered barley is preferred in brewing industry Naked barley is

therefore advantageous to use in food production since no hull

9

needs to be removed and thus al l nutrients are retained In

addition using naked barley for malting has previously been

shown to produce malt with a composit ion and enzyme activit ies

comparable to that of normal malts (Bhatty 1996)

Table 21 Typical chemical composition of barley grain

Component Percent Component Percent

Starch 63-65 Lipids 2-3

Sucrose 1-2 Albumins and globulins 35

Other sugars 1 Hordeins 3-4

Water soluble polysaccharides 1-15 Glutel ins 3-4

Alkali soluble polysaccharides 8-10 Nucleic acids 02-03

Cellulose 4-5 Minerals 2

Adapted from MacGregor and Fincher (1993)

In a study two cult ivars of hull- less barley Scout ( two-

rowed) and Tupper (six-rowed) were uti l ized to prepare f lour and

similarly ground fine-pearled and the pearled grain These three

fractions were used to evaluate physiochemical and functional

(bread making) properties The fractions contained 133-189

10

protein 1 1-21 ash and 08-16 fiber palmitic (160) oleic

(181) and l inoleic (182) were the major fatty acids (Bhatty 1986)

Kiryluk et a l (2000) mil led barley to produce the end-

products f ine and coarse-grained f lours middlings and f ine grits

These products differed in their average contents of β -glucan

total dietary f iber ash and protein This product with a weight

yield of 186 contained 672 β -glucan 2512 total dietary

f iber 2 19 ash and 1583 protein All these values were at

about 50 72 55 and 24 respectively higher than in

dehulled barley

Holtekjolen et a l (2006) observed a strong posit ive

correlation between the β -glucan and the amount of soluble non-

starch polysaccharides (NSP) as well as β -glucan and protein

contents The analyzed hull- less and a typical amylose variety

seem suitable for human consumption where high soluble f iber

and nutrit ive contents are desirable These variet ies contained

high contents of β -glucan soluble NSP protein and lower starch

content and could therefore also be suitable for functional food

products aimed at health benefits and cancer prevention

22 Role of dietary fiber

Different countries and research groups have adopted

different definit ions for dietary f iber which has led to

inconsistent results Therefore a committee was formulated by the

American Association of Cereal Chemists (AACC) to evaluate the

definit ions and methodologies used An updated definit ion was

prepared by this committee in 2001 which concluded that ldquoDietary

f iber is the edible parts of plants or analogous carbohydrates that

11

are resistant to digestion and absorption in the human small

intestine with complete or partial fermentation in the large

intestinerdquo (DeVries 2001)

Dietary f iber includes polysaccharides ol igosaccharides

l ignin and associated plant substances and the data regarding the

beneficial effects of dietary f iber more than two decades have

been recorded According to Schneeman (2001) dietary f iber

regulates the rate of nutrient digestion and absorption serves as a

substrate for the microflora of the gut and promotes laxation The

dietary f iber to foods is usually added for improving their

nutrit ional characterist ics (Brennan and Cleary 2005) However

dietary f iber have both physiological and technological

properties and its addition wil l also alter processing and

handling of foods as well as their texture color f lavor and taste

Many reports demonstrating the role and physiological

functioning of dietary f iber in human health and are involved in

reduction in cardiovascular diseases colorectal cancer and blood

cholesterol and glucose level

Intake of total dietary f iber especial ly from cereal and grain

products (Bingham e t a l 2003 Jansen et a l 1999) can act as a

shield against diabetes (Maier et a l 2000 Schulze et a l 2004) I t

also helps in smooth bowl movement (Sanjoaquin et a l 2004) and

it is effective against constipation (Dohnalek et a l 2004) The

foods r ich in dietary f ibre provide low energy to the body and

interfere with absorption of harmful compounds There dietary

f iber also showed to decrease the serum cholesterol levels (Brown

et a l 1999)

12

Water-retention capacity is another important function of

dietary f iber According to their water solubil i ty dietary f iber can

be classif ied in to two grouprsquos i e soluble and insoluble f ibers

Soluble f ibers include mainly gums pectin and mucilage while the

insoluble f ibers include cel lulose hemicelluloses and l ignin

(Izydorczyk et a l 2002) Barley β -glucan which is soluble dietary

f iber can successfully be used in food system

23 β -glucan Sources and occurrence

The term β - (1rarr3)-D-glucan includes a very large number of

polysaccharides from bacterial fungal and vegetable sources

Their structures have a common backbone of β - (1rarr3) l inked

glucopyranosyl units but the polysaccharidic chain can be β-(1rarr6)

branched with glucose or integrate some β -(1rarr4) l inked

glucopyranosyl units in the main chain (Brennan and Cleary

2005)

The barley crop is used for human consumption due to the

presence of i ts functional ingredients Among al l the cereals

barley and oat are famous for β-glucan Mixed-l inkage (1rarr3)-

(1rarr4)-β-D-glucan or β -glucan is the most abundant component

of the soluble dietary f iber in both oats and barley I t is a l inear

and partial ly water soluble polysaccharide that consists only of

glucose I t is a soluble f iber component found predominantly in

other cereal crops The (1rarr3)-(1rarr4)-β -D-glucan is cel l wall

polysaccharide of cereal endosperm and aleuronic cel ls

Environmental conditions seem to exert a signif icant effect on the

β -glucan content of the cereal grain (Aastrup 1979)

13

β -glucan is one of the minor constituents in barley grains I t is

primarily associated with genotype and is s ignif icantly affected

by the environmental conditions There is a variation in barley β -

glucan content between different locations as documented by

Aman et a l (1989) Zhang et a l (2002) determined and extracted

β -glucan content of barley cult ivars collected from various areas

of China as well as from Canada and Australia by an enzymatic

method For 164 cult ivars originating from China β -glucan

content ranged from 298 (Sumei 21) to 862 (QB25) with a

mean of 4 58 Ragaee et a l (2001) also demonstrated that the

primary sources of β -glucan in the human diet are oats barley

rye and wheat The levels of β -glucan in dehulled or naked oats

and most dehulled or naked barleys range mostly from about 3

to 7 (Lee et a l 1997) in rye about 2 and in wheat less than

05 (Beresford and Stone 1983)

The structures of β -glucan in barley and oat are different

(Wood 1994) Barley β -glucan was found to contain one quarter β -

(1rarr3) l inked units whereas oat β -glucan contained

approximately one third The oat β -glucan structure therefore

contains more β -(1rarr3) l inkages than barley β -glucan (MacGregor

and Fincher 1993) The oligosaccharide with DP3 i e 3-O-β -

cel lobiosyl-D-glucose is the main product and DP4 i e 3-O-β -

cel lotriosyl-D-glucose comes second These two constitute over

90 of the total β -glucan content (Wood et a l 1994) For

structural differences of β-glucan often DP3DP4 ratio is used as

indicator (Izydorczyk et a l 1998a) According to many authors

this ratio is lower for oat than for barley β -glucan Structural

differences have also been reported to exist between soluble and

14

insoluble β -glucans with the ratio DP3DP4 being higher for

insoluble than for soluble β-glucans (Izydorczyk et a l 1998b)

24 Extraction of β -glucan

Various techniques for the isolation of βndashglucan have been

developed β -glucan from barley and oat could be isolated by dry

mill ing and solvent extraction (Wu et al 1994 Dawkins and

Nnanna 1993 Saulnier et al 1994) Among both isolation

methods about 89 βndashglucan could be recovered by solvent

extraction and only 31 by dry mill ing and air classif ication (Wu

et al 1994) from barley and oat However 41-81 βndashglucan on

dry matter basis could be extracted by using neutral or an alkaline

medium (Burkus and Temell i 1998) Furthermore more than 90

extraction could be achieved by hot water extraction (Morgan et

al 1998)

Bhatty (1995) compared different solvents for the extraction

of β -glucan from one sample of hull- less barley bran and revealed

that sodium hydroxide was the most eff icient solvent for

extraction The extraction with sodium hydroxide removed 84 of

the β -glucan compared to 72 by sodium carbonate solution and

only 61 by sequential extraction with water at 40 65 and 95degC

The amount of β -glucan is an important factor in considering

health ef fects In the isolation processes some β -glucan may be

lost Thus the total β -glucan content can not be determined from

the isolated β -glucan (Rimsten et a l 2003) The most frequently

used method for β -glucan determination is i l lustrated by

Association of Official Analytical Chemists (AOAC 1995) This

method involves the dissolution of β -glucan in a buffer

15

hydrolysis with the l ichenase enzyme to ol igosaccharides and

with β -glucanase to glucose Glucose is then analysed

spectrophotometrical ly as a colored substance obtained with an

oxidaseperoxidase reagent (Lambo et a l 2005)

Burkus and Temeil i (1998) have reported that extraction

conditions such as pH and temperature profoundly affect the

viscosity of solutions prepared with β -glucan concentrates I f a

higher concentrat ion of β -glucan is desired in a product low

viscosity extracts may be uti l ized (Burkus 1996)

Carr (1990) explored an improved method for the

determination of (1rarr3)-(1rarr4)-β -D-glucan in cereals and their

products The method includes refluxing of 80 (vv) ethanol to

remove sugars and inactivate of enzymes prior to extraction with

water at 100ordmC for soluble β -glucan determination For several

different food products soluble β -glucan content ranged from

049 to 390 whereas total β -glucan content ranged from 058 to

886 (dry weight basis) The dietary f iber ranged from 48 to

220 for the products

Extraction conditions also determine the properties of

extracted β -glucan Wood et al (1977) extracted the β -glucan gum

pellets through alkali extraction method from oats (Avena sat iva

L) The researchers found that various condit ions such as

temperature pH and ionic strength of the extraction media

affected the β -glucan yields βndashglucan could also be extracted by

using dist i l led water and 4 sodium hydroxide All treatments

differ in their yield and physiochemical properties Extracted

conditions have a great bearing on viscosity properties of β -

16

glucan excessive boil ing during extraction resulted in low

viscosity β -glucan Stable barley β -glucan gum with high viscosity

can be obtained using suitable combination with high pH

(Johansson et al 2000) Recently another method was developed

by Izydorczyk et al (1998) for the extraction of β -glucan through

sequential extraction with water Ba(OH)2 Ba(OH)2H2O and

NaOH In this method each barley sample was extracted 2ndash3 t imes

and the isolated material was combined

The βndashglucan extraction methods for pilot plant levels have

been developed that includes refluxing with 75 ethanol for four

hours prior to extraction-deactivated glucan The pilot plant

extracted gum has less viscosity than bench gum this is due to

high shear rates enzyme activity of fungi and bacteria in pilot

plant conditions (Wood et al 1989) The foods containing βndash

glucan needs viscosity stabil i ty for increased shelf l i fe In another

study i t is found that i f 1N sodium hydroxide is used for βndash

glucan extraction from barley and oat i t affect βndashglucan activity

(Bhatty 1995) The enzymes (glucanase) present naturally or

produce from microorganisms and it is investigated that

enzymatic hydrolysis create problem during production and food

application Scientists noticed higher activity of endo (1rarr3) β -D-

glucanase than endo (1rarr3) (1rarr4) β-D-glucanase (Brunswick et al

1987) Similarly steaming and kilning inactivate l ipases of barley

microbial enzyme are more heat stable than the endogenous

glucanases (Balance and Meredith 1976 Wood et al 1989)

Similarly a method of pure β -glucan extraction has been

provided by Westerlund et a l (1993) and this method involves

defatt ing with propan-2-ol ( isopropanol IPA) and petroleum

17

ether dissolution in water at 96 degC and hydrolysis of starch with

heat-resistant α -amylase The polysaccharides are precipitated

with 60 ethanol at 4 degC and the precipitate is dissolved in water

The solution is treated with 30 (NH4)2SO4 which specif ical ly

precipitates β -glucan but leaves arabinoxylans in solution The

precipitate is dissolved in water and dialyzed against water at

room temperature

25 Health benefits of β -glucan

Barley grain bas been shown to be an excellent source of

both soluble and insoluble f iber and according to dieti t ians and

health professionals i t should be extensively used in diets to

improve health (Oscarsson et a l 1996) During the last 10 years

studies have identif ied a low glycemic-index (GI) diet as

beneficial in relation to the insulin-resistance syndrome Several

semi-long-term dietary interventions are available for healthy

subjects and for subjects with metabolic diseases With a few

exceptions these studies have shown that a low-GI diet not only

improves certain metabolic consequences of insulin resistance but

also reduces insulin resistance per se (Del Prato et a l 1994) In

addition to improvements in glucose and l ipid metabolism

(Jenkins et a l 1987 Brand et a l 1991 Jarvi et a l 1999) there are

indications of improvements in the f ibrinolytic activity (Jaumlrvi et

a l 1999) suggesting a beneficial role in diabetes and

cardiovascular disease I t has been est imated that a 3 85 unit

reduction in GI can be perceived per gram of β -glucan f iber in a

50 g carbohydrate portion of food The viscosity of the f iber

relates posit ively to the degree of f lattening of postprandial

glycemia (Wood et a l 1994 Jenkins et a l 1978)

18

The potential physiological mechanisms behind the eff icacy

of β -glucan are suggested to be i ts abil i ty to retard the absorption

rate of food in the intestine due to increased viscosity in this way

balancing the post-prandial glucose and insulin response (Wursch

and Pi-Sunyer 1997 Wood et a l 2000) In addition some

investigators (Gallaher and Hassel 1995 Jal i l i et a l 2000) has

reported an increased viscosity in the small intestine which may

interferes with cholesterol absorption or re-absorption in this

way affecting the cholesterol balance and synthesis in the body

Therefore i t would be interesting to investigate what kind of

effect could be achieved with general information about the

dietary f iber content (Stone and Clark 1992)

Another physiological aspect with reference to β -glucan was

experienced in intestinal tract that i t s low down glucose

absorption and therefore regulate blood glucose (Wood et a l

1990 Wood et a l 1994) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property may be useful

in the formulation of products targeting management of diabetes

The mechanism by which β -glucan lowers blood glucose and

cholesterol levels may be related to i ts viscosity bi le salt binding

capacity or ferment abil i ty (Davidson and McDonald 1998

Marlett et a l 1994) The enrichment technique and water

extractionfreeze drying technique could enable the use of barley

as a source of a high-value f iber for reducing the glycemic index

of tradit ional wheat-based foods such as bread without affecting

their sensory characterist ics (Cavallero 2002)

β -glucan incorporated functional food tends to reduce

glycemic indices while maintaining palatabil i ty (Jenkins et a l

19

2002) β -glucan containing food bars have an intermediate

glycemic index of 78 (Foster-Powell and Miller 1994) Enrichment

with additional β -glucan is required in order to produce a low

glycemic index barley product (Tappy et a l 1996) which could

also have an increased hypocholesterolemic effect (McIntosh et a l

1991)

Dongowski et a l (2002) reported that diets containing more

soluble macromolecular dietary f ibers such as β -glucan affected

the excretion of bi le acids and neutral sterols the most whereas

the fermentation of dietary f iber including resistant starch

influenced the steroids in feces I t has been hypothesized that

upon ingestion β -glucan increases small intest inal viscosity due

to i ts lower molecular weight and its tendency to form viscous

gummy solutions result ing in reduced bile acid and cholesterol or

tr iglyceride absorption thus lowering plasma cholesterol as well

as altering digestive enzyme activity

More research is in progress to determine the effect of β -

glucan and phytosterols into low-fat spreads and non-fat

phytosterol formulations (Moreau et a l 2002) The cholesterol-

lowering potential of β -glucan and phytosterols may thus depend

upon previous dispersion into a fat matrix and on the physical

nature of the food I t is reported that these compounds have a

capacity to reduce plasma cholesterol concentrations when

consumed in different food matrices but their effect iveness in

non-fat or low-fat beverages has not been established (Jones et

a l 2003) Two mechanisms for serum cholesterol level have been

elucidated in the scientif ic l i terature one deals with the viscous

nature of β -glucan provides a physical barrier that slows down or

20

inhibits the absorption of cholesterol and other l ipid constituents

and second mechanism is about binding of the bi le acids in the

gut The unabsorbed and bound components then proceed to the

large intestine and are excreted from the body Some of the β -

glucan that reaches the colon wil l also undergo fermentation by

colonic microorganisms (Wood and Beer 1998 Casterl ine et a l

1997 Bell et a l 1999) Short chain fatty acids are produced as a

result of fermentation of β -glucan in large intestine

β -glucan have cholesterol lowering action in human body

The cholesterol lowering mechanism involved the suppression of

intestinal cholesterol absorption while partial ly suppressing

cholesterol biosynthesis ( Jones et a l 2000 Plat and Mensick 2001)

only a small part of these are absorbed through intestinal micelle

into blood circulation phytosterol solubil i ty and incorporation

into intestinal micelles is found an important aspect of

phytosterol cholesterol lowering eff icacy Most recent studies

conducted to examine the l ipid-lowering potential of β -glucan

incorporated them into a fat matrix margarine butter or

dressing Results from these tr ials have shown that β -glucan

consumption decreases total cholesterol and LDL- cholesterol

concentrations by 34 to 116 for total cholesterol and 54 to

155 for LDL cholesterol ( Jones et al 2000 Hall ikainen et al

2000 Mussner et al 2002) Oat bran is r ich in β -glucan f iber and

has been shown to lower cholesterol (Anderson et al 1990) This

is bel ieved and found that barley and oat lowers the blood

cholesterol and attenuates postprandial glucose response due to

soluble dietary f iber cal led (1rarr3) (1rarr4)-β -D-glucan also referred

to as β -glucan (Ripsin et a l 1992 Tappy et a l 1996 Drzikova

21

2005) Oat bran reduced total serum cholesterol in

hypercholesterolemic subjects by as much as 23 with no change

in high density l ipoprotein (HDL) cholesterol Since oat bran was

enriched in β -glucan (Wood 1986 Wood et a l 1989) the authors

reported an inverse correlation between serum cholesterol levels

and β -glucan intake Barley and oats are a r ich source of the

soluble f ibre β -glucan which has been shown to signif icantly

lower LDL-cholesterol ( Joseph et a l 2007)

Oat bran providing 73 g β -glucan in a breakfast cereal or 6 2

g in a bar gave signif icantly lower postprandial glucose responses

in NIDDM subjects than an oat bran breakfast cereal providing 37

g and it was calculated that the glycemic index was lowered 4

units for every gram of β -glucan (Jenkins et a l 2002)

In a study different breads were made one from hull- less

barley f lour and the other from two (1rarr3 1rarr4)-β -glucan enriched

fractions The remaining two from a sieved fraction (SF) and a

water-extracted fraction (WF) were produced and evaluated for

sensory evaluation For eff icacy study eight adultsrsquo subjects were

fed test meals of each of the four breads containing the same

amount (50 g) of available carbohydrate and glycemic indices

calculated from finger-prick capil lary blood samples A l inear

decrease in glycemic index was found for increasing (1rarr3) (1rarr4)-

β -glucan content This research confirms the effectiveness of

viscous (1rarr3) (1rarr4)-β -glucan in reducing postprandial blood

glucose levels even in foods with a high glycemic index

(Cavallero et a l 2002)

22

The abil i ty to detect a signif icant effect on glycemic

response related to the dose of β -glucan In a study of the effect of

an oat bran highly enriched in β -glucan (15 dwb) incorporated

into an extruded breakfast cereal subjects with non-insulin-

dependent diabetes mell i tus consumed meals with 4 6 and 86 g

of β -glucan All 3 breakfasts signif icantly decreased the peak and

the average increases in glucose and insulin compared to a

control There was a signif icant relationship between plasma

glucose peak and area under the glucose curve and the amount of

β -glucan in the cereals (Tappy et a l 1996) Wood et a l (1990)

showed that both oat gum and guar gum signif icantly decreased

the postprandial glucose rise Scientists conducted a study and

showed that whole meal bran and f lour from three barley

genotypes which contained graded levels of soluble f iber were

compared with similar commercial fractions of oats for their effect

on cholesterol tr iglycerides high-density l ipoprotein (HDL)

cholesterol and l iver cholesterol ( test model using

hypercholesterolemic rats) Whole meals of the three barley

genotypes contained 30 5 2 or 6 8 soluble f iber oatmeal

contained 30 In meal-fed rats barley genotypes did not show a

favorable blood or l iver l ipid response compared with oats

However in bran- and f lour-fed rats the data showed that

barley exerted a profound blood and l iver cholesterol- lowering

effect compared with oat bran or f lour (blood triglyceride levels

were minimally affected) Blood HDL-cholesterol levels were

appreciably elevated in rats fed barley bran or f lour compared

with oat bran or f lour These results suggested that barley and its

major fractions (bran and f lour) may evoke different l ipidemic

23

responses and that barley bran and f lour have a more favorable

effect on blood l ipids than do oat bran and f lour (Ranhotra et a l

1991)

Wallace et a l (1997) developed product containing high-

fiber high-carbohydrate diets including foods with low glycemic

index have been associated with prevention and treatment of

diseases such as coronary heart disease and diabetes β -glucan a

soluble viscous polymer found in oat and barley endosperm cell

wall was incorporated into pasta test meals Five fasted adult

subjects were fed test meals of barley and durum wheat blend

pasta containing 100 g of available carbohydrate 30 g of total

dietary f iber (TDF) and 12 g of β -glucan or al l durum wheat pasta

containing the same amount of available carbohydrate 5 g of TDF

and negligible β -glucan The β -glucan and durum wheat pasta

resulted in a lower glycemic response as measured by average

total area and maximum increment of the blood glucose curves

Lower insulin response to the β -glucan and durum wheat pasta

was also indicated by lower average area and increment

characterist ics of the insulin curves Barley β -glucan may be an

economical and palatable ingredient for processed food products

formulated to modify glycemic and insulin response

Lia et a l (1995) studied the effect of β -glucan on the

excretion of bi le acids using breads baked with oat bran oat bran

with β -glucanase barley or wheat in the diet of i leostomy

subjects They showed that the excretion of bi le acids was 53

higher with the oat bran bread than with the bread containing oat

bran and β -glucanase and also signif icantly higher than with

barley and wheat bread The excretion of cholesterol was higher

24

for barley bread than for wheat or oat bran-β -glucanase bread In

one of the few studies that have reported MW values a drink

containing 5 g β -glucan of MW 70000 extracted from oat bran

signif icantly lowered postprandial glucose and insulin levels

relative to a r ice drink control whereas a similar drink containing

barley β-glucan of MW 40000 was without signif icant effect

(Biorklund et a l 2005)

A study was further conducted to est imate the glucose

insulin and glucagon responses after consumption of high-soluble

β -glucan compounds from oats and barley The study includes 11

men and 11 women non diabetics between 35-57 years old

subjects Different tests (blood and urine) performed to analyze

the glucose responses The prel iminary results showed the

signif icant decrease in oats barley and both extracts than glucose

solution High-soluble barley f iber is more effective than standard

oats Oat and barley carbohydrate-based fat substitutes can

provide a useful addition to control plasma glucose responses

(Hallfr isch et a l 2003)

Investigations are further continued to f ind the cholesterol-

lowering activit ies of oats and barley In this study the anti

atherogenic properties of β -glucan concentrates from oats and

barley were evaluated in Syrian golden F1B hamsters by

consuming a semi purif ied hypercholesterolemic diet (HCD)

containing cholesterol (0 15 g100 g) hydrogenated coconut oi l

(20 g100 g) and cel lulose (15 g100 g) The experimental diet HCD

formulated with different levels of β -glucan (2 4 or 8 g100 g)

from oat and barley instead of cel lulose In agreement with

previously proposed mechanisms total fecal neutral sterol

25

concentrations were signif icantly increased in hamsters

consuming 8 g100 g barley or oat β -glucan Aortic cholesterol

ester concentrations were signif icantly reduced in hamsters fed 8

g100 g β -glucan from barley or oats From this observational

study found that the cholesterol- lowering potency of β -glucan is

approximately identical whether i ts origin was oats or barley

(Delaney et a l 2003)

26 Functional properties of β-glucan

Other than nutri t ional benefits obtained from β ndashglucan i t

also have valuable functional properties such as thickening

stabil izing emulsif ication and gelation which make β -glucan

suitable for incorporation in soups sauces beverages and other

food products (Dawkins and Nnanna 1993 Burkus and Temell i

1999) Such functional properties are very important for new food

applications However proper knowledge on thermodynamic

properties of βndashglucan in a food system with other food

components is necessary to exploit full benefits (Burkus 1996)

Gelation is associated with cross l inking of long chain of

polymer to form three dimensional continuous networks this

structure traps and immobil izes the l iquid and become thick

enough to f low under pressure (Glicksman 1982) βndashglucan is a

long chain of glucose units counts for 3-7 of total grain weight

which make i t more viscous Both amylose and βndashglucan are

straight chain of glucose I t has been found that amylose chains

al ign themselves and form gel while βndashglucan form gel through

interrupted regions of β -(1rarr3) l inkages (Buliga et al 1986) Due

to presence of glucose bond between (1rarr3) (1rarr4) l inkages that

26

make barley βndashglucan a soluble f iber β -glucan provides excellent

viscosity forming properties and used as thickening agents in

different food applications e g salad dressings sauces and ice

creams (Wood 1986) Thus addition of barley β -glucan into foods

not only to give better nutrit ional enhancement but also help to

improve quality parameters such as processing behavior and

shelf- l i fe or stabil i ty ( Klamczynski and Czuchajowska 1999)

Thammakiti et a l (2004) determined and evaluated that β -

glucans obtained from spent brewers yeast and its potential food

applications The objective of the study was to evaluate the effect

of homogenization on the rheological properties chemical

composit ion and functional properties of β -glucan In case of

homogenized cel l walls higher β -glucan content and apparent

viscosity has been observed than those which had not been

homogenized due to the breakup of cel l walls This extracted β -

glucans has shown higher apparent viscosity water-holding

capacity and emulsion stabil izing capacity but very similar oi l -

binding capacity when compared with commercial β -glucans from

bakers yeast

Dawkins and Nnanna (1995) reported that β -glucan viscosity

and stabil i ty showed diverse behavior when maintained different

pH-temperature-time combinations during processing and

decrease stabil i ty of food systems such as salad dressings i f β -

glucan is used as a stabil izer The presence of other food

ingredients can affect properties of hydrocolloids Sweeteners

alter the solution properties such as sucrose in low to mild

concentrations increased viscosity of oat β -glucan while higher

concentrations lowered viscosity Similarly Beer et a l (1997) has

27

substantiated that processing may affect solubil i ty of β -glucan

and decrease the molecular weight of β -glucan I t is obvious that

when β -glucan is used in bread making signif icant

depolymerization of l inear bond of this polysaccharide was

caused (Andersson et a l 2004)

Lyly et a l (2004) conducted a research study on two

different β -glucan sources and found that the sensory

characterist ics of soups prepared from barley β -glucan were

different compared to oat β -glucans Freezing had no remarkable

effect on the molecular weight of β -glucan or on the sensory

attr ibute of the soups The researchers visualized that barley β -

glucan addition resulted in alterations of a foods functional

properties such as viscosity More stable foams and emulsions

were obtained with incorporation barley β -glucan than oat β -

glucan Morgan et al (1998) also observed that βndashglucan from

barley makes soft gel on cooling at more than 05 concentrations

βndashglucan stabil i ty is dependent on t ime temperature and pH

values and these factors affects both viscosity and stabil i ty when

used in foods as stabil izers (Burkus and Temell i 1999) There are

reports by researchers showing that viscosity is a function of

molecular weight I t is important to determine precise molecular

weight to est imate βndashglucan characterist ics for potential

applications into food products Among cereals barley and oat

showing high concentrations of β ndashglucan this unique property

differentiate them from others (Burkus 1996) I t is well known

that barley and oat β -glucan is very similar in structure As for as

viscosity is concerned it has been observed that oat β - glucan has

high viscosity than barley due to long molecular chains (Beer et

28

al 1997) Temperature is responsible for changes in viscosity and

according to observations found that oat β ndashglucan gum viscosity

r ises from 25-370C and start decreases from 610C and maximum

reduces at 1000C when compare with control treatment at 250C

(Dawkins and Nnanna 1995) Furtehrmore barley βndashglucan

imparts a smooth mouth feel to beverage products while also

making the beverage an excellent source of soluble dietary f iber

In beverage formulations i t can provide similar functionality l ike

other thickeners β -glucan gums have shown such types of results

that are comparable with other thickners such as alginates pectin

xanthan and carboxymethylcel lulose (Giese 1992)

27 Utilization of β -glucan in food products

Food industry has a major focus on the production of foods

containing health-enhancing components that wil l improve

consumer health beyond meeting basic nutrit ional requirements

(Sloan 1999) Currently functional and nutraceutical ingredients

are used to exploit their health benefits and it has been found that

beverages provide excellent medium for their addit ion (Kuhn

1995) Barley is suitable for a range of food applications and it can

be processed into a number of palatable and nutrit ious food

products As other polysaccharides β -(1rarr3)-D-glucans have

found a very large range of possible applications in various

industries and especial ly in foods cosmetic agronomy

therapeutic and other In food industry beside typical

applications of polysaccharides as thickening agent and

stabil izers β - (1rarr3)-D-glucans have an increasing interest in the

areas of edible f i lm and wide application into feed for domestic

animals and low calorie food as chemical additives are not famous

29

among the consumers Barley gives r ise poor baking quality and

also not having good taste and appearance aspects which have

l imited i ts use in human foods However in current years there

has been an increasing research interest for the exploitation of

barley in a wide range of food applications (Bhatty 1999)

During the last few years functional drinks sector has been

strong and expected to continue Growth in future (Potter 2001

Sloan 2002) Industry analyst predict and saying continuous

growth and latest research has focused on the use of soluble

dietary f ibre and in particular cereal β -glucans as stabil izers in

the manufacture of low-fat products such as salad dressings

(Kontogiorgos 2004) ice creams yoghurts (Brennan 2002) cheese

and many other food products The use of β-glucans preparation

to partial ly substitute vegetable oi l in the formulation and is

found that give us many advantages in the food system Barley β -

glucan is a compound which as attractive thickening properties

and does not reveal deteriorative changes during processing and

storage periods I t gives r ise good thick solution properties when

added into water I t is suggested that β -glucan gum can be used

as thickener in different food application i e in ice cream sauces

and salad dressing (Carr et al 2002) Furthermore no bad effect on

sensory properties was reported There is an est imate and

predictions by industry analyst that functional drink wil l make a

good share in food section (Sloan 2002)

Erkan et a l (2005) produced tarhana (fermented cereal

product) samples from hulless and hulled barley with relatively

high β -glucans content Chemical and sensory properties of the

tarhana samples were examined and evaluated with the

30

tradit ional wheat tarhana During fermentation some of the β -

glucans may be destroyed however the results indicated that

barley f lours can be uti l ized to produce tarhana with relatively

high β -glucans content Effect of tarhana production on the

electrophoretic properties of proteins was est imated in this study

by using SDS PAGE Relative band intensit ies of tarhana samples

were generally less intense than those of respective f lour samples

perhaps due to the hydrolysis of proteins during fermentation

However the overall sensory attributes showed that uti l ization of

barley f lours in tarhana formulation resulted in acceptable soup

properties in terms of most of the sensory properties

Another product where Barley has been effectively

incorporated by (Sidhu et a l 1990) and made single layer f lat

breads including chapatis and Turkish bazlama bread by Basman

amp Koksel (1999) A further study conducted by Berglund et a l

(1992) and he has successfully used hull- less barley f lour in

chemically leavened products such as biscuits pancakes muffins

and cookies Such yeast- leavened bread made with hull- less

barley f lour is also being a good dietary source of (1rarr3) (1rarr4) β -

glucan Tradit ionally barley is not often used in bread products

because i t is deficient in gluten and has poor sensory qualit ies

Izydorczyk et a l (2001) showed that barley might replace up to

20 of wheat f lour without causing too much disturbance to the

overall dough quality

Similarly Morin et a l (2002) established that addition of

barley β -glucan gum (762 purity) into reduced-fat breakfast

sausages to such an extant that i t provides 03ndash07 β -glucan in

31

the manufactured goods gave better water binding and at a level

of 0 3 having no signif icant effects on product texture or f lavor

A study performed by Volikakis et a l (2004) in which he

used elevated level of β -glucan in cheese A commercial

concentrate of oat β -glucan (222 β-glucan content) has been also

incorporated into low-fat white-brined cheese from bovine milk

(70 fat reduction) at two levels 0 7 and 14 (ww) This

product showed in an increased yield greater proteolysis and

higher levels of short chain fatty acids ( lactic acetic and butyric)

as well as with improved texture compared to i ts low-fat (β -

glucan-free) counterpart However the product made with the

high level of β -glucan has shown signif icantly inferior impression

scores for colour f lavour than those of a typical white-brined

cheese product

28 Physico-chemical characteristics of beverage

Among functional foods beverages have excellent

opportunit ies for the incorporation of nutraceutical ingredients

Giese (1992) stated that the new formulations of beverages are

rapidly changing The market shelves are full of different

beverages with not only soda pop juices and dairy beverages

There is huge number of food products taken as beverages such as

iced teas and coffees sports drinks herbal teas frozen carbonated

beverages mint blends vegetable juices smoothies Soft drinks

have tradit ionally remarkable share in the market However in

current years consumers have not been choice for tradit ional

drinks but also have more exotic beverages such as the teas iced

coffees isotonic or sports drinks and non-carbonated beverages

32

and ready-to-drink iced herbal teas are also gaining popularity

(Swientek 1998)

Beverages not only provide taste and refreshment

satisfaction but can also offer a ready and unique delivery system

for protein vitamins minerals and other food ingredients such as

dietary f iber A major challenge to develop a nutraceutical

beverage is to preserve i ts nutrients and to make i t taste good

Another challenge involves the processing of these beverages with

minimum losses of f lavor vitamins and color Barley β -glucan is

being used frequently in cereal products According to FDA new

types of foods containing β -glucan are need to promote in which

3g of β -glucanday should be used this is the amount defined

amount to get the potential health effects Beverages showed

suitable category for new product development containing β -

glucan as functional ingredient

FDA has recommended consumption of 3 g β -glucan per day

to achieve such health benefits This claim was amended later on

and includes oat extracts containing up to 10 βndashglucan (FDA

2002) Some studies showed that consumers want to pay more for

foods having functional benefits ( Jonas and Beckmann 1998)

Processing condit ion for extraction of β -glucan is important

because i t may affect physiological molecular weight and

solubil i ty of barley βndashglucan (Beer et al 1997) and therefore has

influence on i ts physiological eff icacy and products development

High molecular weight β -glucan is particularly sensit ive to

processing Freezing has not been found to affect the molecular

weight of β ndashglucan (Suortt i et al 2000 Kerckhoffs et al 2003)

but i t decreases the solubil i ty of βndashglucan (Beer et al 1997) On

33

the other hand heating makes β-glucan more soluble (Bhatty

1992 Jaskari et al 1995) and enhances i ts physiological eff icacy

The beverage prepared at high temperature had a sl ightly

higher apparent viscosity than the pulse electric f ield (PEF)

treated beverage and developed sedimentation problem in the

container during storage The PEF processed beverage maintained

its natural orange juice l ike color was better than the heat treated

beverage which developed a sl ightly whitish color However the

PEF treated product was less microbiological ly stable at

refrigeration temperature compared with the heat treated product

which was stable for more than 12 month (Sharma et a l 1998)

Temell i e t a l (2004) prepared an orange-flavored barley β -

glucan beverage with different β -glucan levels and compared with

same level pectin beverage and analyzed for different sensory

parameters and the trained panelists found peely and fruity

orange aroma and sweetness intensity to be similar for al l

beverages tested Beverage sourness intensity differed among

beverages Panelists evaluated beverages containing 03

hydrocolloid as similar whereas beverages with 05 and 07 β -

glucan were more viscous than those with pectin at these levels

Acceptabil i ty of beverages was similar according to the consumer

panel During the f irst week of storage Colorimeter values of

beverages decreased mostly stabil izing thereafter With an

increase in concentration β -glucan beverages became l ighter in

color and cloudier but these attr ibutes for pectin beverages were

not affected During the f irst three weeks of storage β -glucan

beverages exhibited cloud loss

34

Barley β -glucan has revealed beneficial nutrit ional and

physical functionality characterist ics that are required for

beverage making (Temell i et al 2004) β -glucan can be used in

combination with whey protein isolate (WPI) for functional

beverage development This beverage has shown good results for

quality overall acceptabil i ty and remained acceptable for 8-week

storage Non-signif icant results for other quality parameters such

as sweetness sourness and f lavor intensity was observed Many

researchers have attempted the use of βndashglucan in beverage

(Holsinger et al 1974 Pendergast 1985) Whey protein in

combination with βndashglucan is successfully using in other food

systems due to nutrit ional and functional properties Different

diseases can be prevented with the help of barley βndashglucan and

whey protein isolates when used in foods (Temell i et al 2004) βndash

glucan is extracted from oats and oat porridge is made after

consumption it was demonstrated that product has reduce

postprandial blood glucose level (Wood et al 1990 Wood et al

1994) These developments led top the approval of a health claim

for oats by the Food and Drug Administration (FDA) in the United

States indicating that oatmeal whole oats and oat products

containing 075 g of β -glucan per serving may reduce the risk of

heart disease FDA 1999) Kulkarni et al 2008 made a barley tea-

l ike extract that is a popular summer drink in Japan and explained

the effects of various temperatures between 1500C and 2800C

during sub crit ical water extraction of barley Each barley extract

was carried out for antioxidative activity amount of residual

matter and sensory properties that were found at 2050C I t was

found that 5-Hydroxymethyl-2-furaldehyde is the most important

antioxidative component of the extract at 205oC

35

Many researchers worked on soft drinks and beverages and

conducted different analysis on quality parameters as DrsquoHeureux-

Calix and Badrie (2005) observed the color and microbial aspect of

puree during storage At pH 23 an intense red color is achieved

There were no signif icant changes observed for physicochemical

parameters except consistency and hue angle for color The puree

contained the total soluble solids in the range of 410ndash435degBrix

and pH was 262 There are reports for the development of new

formulations and then undergo sensory evaluation process to test

their consumer acceptance Maestri et a l 2000 added the ethylene

diamine tetra acetic acid (EDTA) in soy bean and proposed a new

method to attain a soybean with improved f lavor characterist ics

and found that a waterbean ratio of 4 5 1 has given better

results and provided the best protein (422 g 100 ml- 1 ) and total

sol ids (880 g 100 ml- 1 ) contents The soybean was evaluated for

pH viscosity and density as well as for protein compare with

soybean beverage

In the same way Singh and Nath (2004) test i fy different

composit ions for beverage and used denatured whey protein

concentrate (WPC) in the presence of pectin and carboxy

methylcel lulose (CMC) The formulation of beverage was 25 bael

fruit pulp 16degBrix and pH 39 and was fort i f ied with 175 2 75

and 375 level of WPC-polysaccharide complex Among al l

combinations he rated foodstuffs with 175 protein level of

pectin-WPC complex and 175 and 275 protein level of CMC-

WPC complex Moreover 1 75 whey protein level of CMC-WPC

complex was assigned maximum scores for al l sensory aspects

36

Lakshmi et a l (2005) optimized the conditions for beverage

formulations They used mixture of enzymes varying pH

temperature etc under controlled conditions The carbonated

beverage having 125 juice 16degB total soluble solids (TSS) and

04 acidity was suitable for storage During storage beverage

tends to retain i ts quality attr ibutes l ike taste and f lavor up to 2

months Refrigeration of the produce could be imperative in

enhancing the shelf l i fe of the produce Refrigeration at colder

temperatures also favors the retention of active components as

Prati et a l 2004 revealed ascorbic acid content maintained their

level during storage with a loss of only 20 in relation to the

concentration added

Different combinations used by Suh et al 2003 including

barley sprouting and sweet potato The mixture of barley sprouts

and sweet potato was uti l ized in the ratio (11) to increase the

industrial applications of sweet potato and rice beverage I t was

also established that the heat stabil i ty of amylase in sweet potato

is higher than that in barley Reducing sugar content in the

mixture of barley sprouts and sweet potato was higher than in

either barley sprouts or sweet potato alone Sahu et a l 2005 used

lemon grass in beverage formulations and observed that fresh

beverage having 152degB total soluble solids (TSS) pH 435 2329

total sugars 4 53 reducing sugars 0 19 acidity and 15 lemon

grass dist i l late obtained the average sensory score of 8 58 which

was highest among the other beverages prepared with different

concentrations of lemon grass dist i l late At small scale barley and

pectin beverage can be produce by adding water in steam jacket

kett le then mix βndashglucan or pectin and boil for one minute

37

sucrose is premix in water This whole mixture is cool down to 70 oC Add High fructose corn syrup and orange f lavour then

homogenize at 2000 psi shift mixture into steam kett le and add

ascorbic acid ci tr ic acid and βndashglucan The mixture is Pasteurize

at 90oC for half minute At the end bott les are hot f i l led and

placed at refrigerator temperature (Temell i et al 2004)

Barley (Hordeum vulgare L) is mainly used for brewing in

developed countries and as animal feed in less developed

countries However barley has great potential due to soluble f iber

content for human consumption and industr ial uses The cel l walls

of barley grain contain more βndashglucan as compared to aleurone

cel l walls The addition of βndashglucan in water wil l enhance the

viscosity and used as a thickening agent in beverages The action

of this soluble dietary f ibre is just l ike a typical visco-elastic

polysaccharide l ike pectin guar gum carboxymethylcel lulose

(CMC) and xanthan when used in different food products In

recent era the application of βndashglucan in food matrix play a key

role as a functional dietary f ibre

The development of functional beverages by incorporating

βndashglucan show excellent results as a nutraceutical ingredients

Barley βndashglucan gum is stable in low pH conditions and in

refrigerated storage The purity of βndashglucan depends upon

extraction and isolation method used The unpurif ied samples of

βndashglucan causes problem when added in to the food systems The

increasing trend of viscosity due to βndashglucan is considered to be

an important factor in lowering the postprandial blood glucose

levels and cholesterol

38

Distinctive research is mandatory to est imate the effect of

various process parameters on the rheological characterist ics and

molecular weight profi les of βndashglucan extracts and determine how

processing affects the eff icacy of incorporated βndashglucan Such

research would widen our perceptive to know how βndashglucan may

affect the nutrit ional properties of foods by altering their texture

structure and viscosity

39

CHAPTER-3

MATERIALS

AND

METHODS

31 Procurement of raw material

Barley variety (Haider-93) was procured from wheat

research insti tute Ayub Agricultural Research Insti tute (AARI)

Faisalabad

32 Preparation of barley flour

The barley f lour was prepared by grinding barley grains

through UDY cyclone mill (mesh size 20 mm)

33 Analysis of raw materials

The barley f lour was analyzed for proximate composit ion by

fol lowing their respective methods as described below

331 Moisture content

The moisture content of barley f lour was determined in an

oven through drying method (at 105degC) according to the

procedure described in AACC (2000) Method No 44-15A The

moisture content of barley f lour was determined by weighing 2 g

of sample into a pre weighed china dish and drying it in an air

40

forced draft oven at a temperature of 105plusmn5degC t i l l the constant

weight of dry matter was obtained The moisture content in the

sample was determined as given below

332 Crude protein

The barley f lour was tested for crude protein content according

to the Kjeldahlrsquos method as described in AACC (2000) Method No

46-30 Two gram of barley f lour sample was taken into the

digestion tube Twenty mill i l i ters of 98 concentrated sulphuric

acid and 2 tablets of digestion mixture (as catalyst) were added

into the digestion tube The digestion was carried out through

digestion unit t i l l transparent residue contents were obtained and

then after cooling 50ml dist i l led water was added The mixture

was neutral ized with 70 ml of 40 NaOH solution in order to

release gaseous ammonia The neutral ized solution was then

dist i l led through Kjeldahlrsquos dist i l lat ion apparatus The ammonia

l iberated was trapped in 4 boric acid solution containing

indicators (methyl red and ethylene blue) The amount of

ammonia collected was then t i trated against 0 1N sulphuric acid

to a purple end point A blank determination was carried out

fol lowing similar procedure without the test sample The

percentage protein was calculated according to formula given

below

Crude protein () = Nitrogen () x 625

Wt of original flour sample ndash Wt of dried flour sample Moisture () = -------------------------------------------------- x 100

Wt of original flour sample

41

333 Crude fat

The crude fat in each such sample was determined by running

sample through Soxhlet apparatus according to the procedure

given in AACC (2000) Method No 30-25 A sample (3 g) was

weighed into an extraction thimble and extraction carried out in

soxhlet appartus with petroleum ether for 2 hours the previously

heated dried cooled and weighed receive f lask containing oil

were dried in a hot air oven cooled in a desiccator and weighed

The fat content was the difference in weight between the empty

receive f lask and the residual oi l expressed as a percentage of the

sample weight

3 3 4 Crude fiber

The crude f iber content in each sample was est imated

by digesting the fat free samples of barley f lour in 125 H2SO4

fol lowed by 125 NaOH solution as described in AACC (2000)

Method No 32-10 After digestion the sample residue was ignited

by placing in a muffle furnace maintained for 3-5 hours at

temperature of 550-650 degC t i l l grey or white ash was obtained The

percentage of crude f iber was calculated after according to the

expression given below

335 Ash content

Ash is a inorganic residue remaining after the material has

been completely burnt at a temperature of 550degC in a muffle

furnace I t is the aggregate of al l non volati le inorganic elements

Weight loss on ignition Crude fiber () = ---------------------------------- x 100 Weight of flour sample

42

present in a material as i ts oxides The ash content of the barley

f lour was determined according to AACC (2000) Method No 08-

01 The f lour Sample (5 g) was weighed into a previously heated

dried cooled and weighed crucible The sample was charred over

a Bunsen f lame unti l no more smoke was given off and then

transferred into a muffle furnace and heated at a temperature of

550degC unti l i t turned to a completely grey material The ash

content was then cooled in a desicator and weighed The

difference in weight between the empty crucible and crucible with

ash residue expressed as a percentage of the original sample

weight and recorded as ash content

336 Nitrogen free extract (NFE)

The NFE was calculated according to the fol lowing expression

NFE = 100 ndash ( moisture + crude protein + crude fat +

crude f iber + ash)

34 Extraction and purification of β -glucan

β -glucan gum was extracted from barley variety (Haider-93)

by fol lowing the method described by Wood et a l (1978) with

some modifications The barley f lour (50 g) was suspended in 500

ml water pH was adjusted to 10 with Na2 CO3 (20 vw) and

st irred vigorously for 30 minutes at a temperature of 45ordmC The

mixture was centrifuged (Model 3K30 Sigma Germany) at 15000 x

g at 4ordmC for 15 minutes The supernatant was adjusted to pH 45

with 2 M HCL and centrifuged again (20 minutes at 21000 x g

4ordmC) to separate precipitated protein which was discarded The β -

glucan was precipitated by the addition of an equal volume of

43

ethanol (999) to the supernatant with slowly st irring The

precipitate was recovered by centrifugation at 3300 x g for 10

minutes I t was al lowed to sett le overnight at a temperature of 4ordmC

in a refrigerator and the sample was dried in a vacuum drier

(Model DZF 6020 R-A-alpha M) The extracted β -glucan was

stored as pellets in high density polyethylene bags at 50C for

further studies

35 Analysis of β -glucan

The purif ied β -glucan pellets were analyzed for different

chemical parameters as described below

351 Proximate composition

β -glucan pellets were analyzed for moisture crude protein

crude fat crude f iber ash and NFE content according to their

respective methods as described in section 33

3 5 1 Total Dietary Fiber (TDF)

The β -glucan pellets were analyzed for total dietary f iber

contents according to method described in AACC (2000) Method

No32-05 The pellets were dispersed in a buffer solution and

incubated with heat-stable α -amylase at a temperature of 95-100

degC for 35 minutes After cooling the samples (gum pellets) up to

60degC incubated at 60degC for 30 minutes by adding of 100 microl

protease solution Finally these contents were incubated with

amyloglucosidase at 60degC for 30 minutes The f iber contents were

precipitated by the addition of alcohol in 1 4 ratio The contents

were f i l tered and washed with alcohol and acetone A blank was

44

run through entire procedure along with test samples to calculate

any contribution from reagents to residue

352 Soluble Dietary Fiber (SDF)

The soluble dietary f iber content in β -glucan pellets were

determined according to the method as mentioned in AACC (2000)

Method No 32-07 by employing Megazyme Assay Kit The

samples were dispersed in buffer solution and incubated with

heat-stable α -amylase at 95-100degC for 35 minutes After cooling

the samples to 60degC and contents by adding 100 microl protease

solution were incubated at 60ordmC for 30 minutes Finally the

contents by adding amyloglucosidase were incubated at a

temperature of 60degC for 30 minutes The residue after f i l tration

was washed and rinsed with 10 ml water The f i l trate and water

washing was weighed and soluble dietary f iber was precipitated

with four volume of ethyl alcohol The contents were f i l tered and

dried and corrected for ash and protein contents A blank was also

run simultaneously through entire procedure along with test

samples to calculate any contribution from reagents to the

residue

353 In-Soluble Dietary Fiber (IDF)

The soluble dietary f iber (IDF) contents in β -glucan pellets

were determined according to the procedure described in AACC

(2000) Method No 32-20 The samples were dispersed in a buffer

solution and incubated with heat-stable α -amylase at a

temperature of 95-100degC for 35 minutes The samples (gum

pellets) after cooling up to 60 degC incubated by adding 100microl

protease solutions at 60 degC for 30 minutes and then the contents

45

were incubated by adding amyloglucosidase at 60degC for 30

minutes The residue after f i l trat ion was washed and rinsed with

10 ml water The resultant residue was weighed and in soluble

dietary f iber was precipitated with four volume of ethyl alcohol

The contents were f i l tered dried and corrected for ash and

protein contents A blank was also run simultaneously through

entire procedure to calculate any contribution from reagents to

residue

354 Pentosans

The pentosans of β -glucan pellets were determined by the

method as described by Hashimoto et a l (1987) The powdered β -

glucan pellets were hydrolyzed with HCl (2N) at a temperature of

100 oC Then after cooling and neutral ization sugars were

removed by incubating through the addition of yeast for 2 hours

and centrifuged at 1000g A mixture of supernatant (2 ml) water

(1 ml) FeCl3 (3 ml) and orcinol (0 3 ml) was vortexed and then

heated for 30 minutes and cooled The absorbance was measured

through spectrophotometer (IREMCO Model 2020 Germany) at

670 nm

3 5 5 Starch

The starch content in β -glucan pellets was determined

according to method described in AACC (2000) Method No76-11

The f inely ground pellet samples were moistened with ethanol

(80) to aid dispersion Thermo-stable ά -amylase was added and

st irred vigorously on vortex mixer The mixture was incubated for

6 minutes at a temperature of 50oC with occasional shaking

Sodium acetate buffer and amyloglucosidase were added and the

46

mixture was st irred and incubated at 50 o C for 30 minutes The

contents were transferred from the tube to 100 ml volumetric f lask

and adjusted the volume by disti l led water The al iquot of this

solution was centrifuged at 3000g for 10 minutes Transferred

duplicate al iquots (01 ml) of the diluted solution to the bottom of

tubes GOPOD (glucose oxidase peroxidase) reagent was added to

sample mixture and blank and incubated these contents at a

temperature of 50oC for 20 minutes The absorbance of test

samples glucose control and blank was measured through

spectrophotometer (IREMCO Model 2020 Germany) at 510 nm

36 Utilization of β -glucan in beverage

The purif ied β -glucan was uti l ized in different formulations

for the preparation of functional beverages The formulation of

treatments is presented in Table 31

Table 31 Treatment plan

Treatments β -glucan ()

T1 0 control (0 2 pectin)

T2 02

T3 04

T4 06

T5 08

T6 10

47

37 Preparation of Barley Beverage

The β -glucan beverage was prepared with some

modifications in the formulation given by Temell i et a l (2004)

The actual composit ion of beverage is given in Appendix I The

f low diagram of beverage preparation is given as under

Fig 31 Preparation of β -glucan

Heat water to 90 o C

Add slowly β -glucan in solution form

Mix by using high speed mixer

Add remaining ingredients according to Formulation

Adjust pH to 32 with acidulant

Thermally processed and f i l l ing in pre steri l ized bott les

Storage at 5oC

38 Analysis of beverage

The β -glucan beverage was analyzed for different

physicochemical microbiological and sensoric attr ibutes

according to their respective methods during three months

storage at 5oC on fortnightly basis The description of methods is

given below

48

381 Color

The color values of β-glucan beverage samples were

measured according to method of Yu et a l (2003) by using the L

a b color space (CIELAB Space) with Color Tech-PCM (USA)

The L Value indicates l ightness the a and b values are the

chromaticity coordinates (a from red to green b from yellow to

blue)

382 Acidity

The acidity of beverage samples was determined by

fol lowing the method given in AOAC (1990) A sample of 5 mL

from each treatment was t i trated against 0 1 N sodium hydroxide

solution to a persistent pink color end point by using two or three

drops of phenolphthalein indicator The results are expressed as

percent citr ic acid and calculated by the fol lowing formula

mL of NaOH times normality of NaOH times eq wt of acid Acidity () = - - - - - - - - - - - - - - - - -- - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - Volume of sample times 10

383 pH

The pH of beverage samples was estimated according to the

method described in AOAC (1990) The samples were taken in a

neat and clean 50 mL beakers and pH was directly recorded by

using a cal ibrated pH meter ( inoLab pH 720 Germany)

384 Total soluble solids

Total soluble solids of functional beverage were recorded by

using hand refractometer equipped with a percent scale and the

results were expressed as percent soluble solids o Brix

49

385 Specific gravity

The specif ic gravity was determined by fol lowing the

method given in AOAC (1990) Empty pycnometer was weighed

and f i l led with water at 20 oC and again weighed Then washed the

pycnometer and dried in oven and weighed again Now it was

f i l led with test beverage sample and weighed At the end specif ic

gravity was calculated by the formula given under

S - E Density of sample = W - E

Where

S = Weight of sample f i l led pycnometer

E = Weight of empty pycnometer

W = Weight of water f i l led pycnometer

386 Viscosity

The viscosity of functional beverages was measured by

fol lowing the procedure of AACC (2000) through Rion viscometer

(Rion Tech USA) after every fortnight interval during the storage

of three months

387 Sugars (Reducing and Non-reducing)

The total sugars (Total sugars reducing sugars and non

reducing sugars) in the beverage samples were est imated by using

the method of Lane and Eynon as described by Ruck (1963)

Fehlingrsquos solution was made by mixing CuSO4 and alkaline

tartrate solution in equal volumes The pure sucrose sample

prepared in HCl was f i l led into the burette and run into the f lask

50

containing 10 ml Fehlingrsquos solution almost whole volume of the

sample as calculated in the incremental method so that less than

05 ml or more than 1 ml was needed to complete the t i tration The

contents in t i tration f lask were boiled after addition of 2 drops of

methylene blue indicator upto brick red end point The 10 ml

Fehlingrsquos solution equivalent was derived in terms of invert sugar

content and found to be 0505g 25 ml beverage sample was taken

into a 400 ml beaker to which 100 ml water was added and

neutral ized with 1 N NaOH The volume was made up with

dist i l led water up to 250 ml and f i l tered with Whatman fi l ter

paper 2 ml of lead acetate solution was added shaken well and

after 10 minutes 21 ml potassium oxalate solution was added and

f i l tered (f i l terate a)

3871 Reducing sugar

The f i l trate (a) was employed for determination of reducing

sugars by standard method of t i tration as described above The

reducing sugars were calculated according to the expression given

below

Fehlingrsquos solution factor x 100 x dilution Reducing Sugars = ----- - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - Volume of sample used

3872 Total sugars

50 ml f i l trate (a) was taken into a 250 ml f lask 5 g citr ic acid

and 50 ml water were added The solution was boiled gently for

10 minutes to invert the sucrose and cooled I t was transferred to

a 250 ml volumetric f lask and neutral ized using phenolphthalein

as an indicator NaOH (20) was added unti l solution turned to

51

pink then 1N HCl was added unti l pink color disappeared The

total sugars were calculated using the fol lowing formula

Fehlingrsquos solution factor x 100 x dilution Total sugars () = - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - - - - - -- - - - - - - - - Volume of sample used

3873 Non-Reducing Sugar

Non reducing sugars were determined according to the

formula given below

Non reducing sugars ()= ( Total sugars()- Reducing

sugars()times 095

39 Total plate count of beverage samples

Total account of microorganisms in beverage was carried out

fortnightly during storage of three months by adopting the

method of (Lateef et a l 2004) as given bellow

391 Preparation of media

Amount of media to be prepared was determined by

deciding on number and frequency of tests and frequency of

making media 23g powdered nutrient agar was added to 1000 ml

of dist i l led water and heated to prepare nutrient agar media

While Sabouraud dextrose agar media was prepared by mixing

dextrose 40 g peptone 10 g and agar 35 g in 1000 ml dist i l led

water and heated

392 Sterilization and incubation of media

The media were steri l ized in autoclave at 15 to 20 Ib

pressure for 15 minutes then these were stored in refrigerator The

52

prepared media were poured in petri dishes and 15 ml of molten

media was also poured in each dish Dilution and media were

mixed by swirl ing the pteri dishes to and forth and al lowed to

solidify and then Petri dishes were inverted to avoid condensation

of moisture inside the cover These petri dishes were incubated at

37oC for 48 hours After incubation period colonies developed in

Petri dishes were counted through Qubec colony counter

310 Sensory evaluation

The functional beverages were organoleptical ly evaluated

for sensory parameters such as colour taste f lavour and overall

acceptabil i ty by a panel of f ive judges The nine point hedonic

scale was employed for the evaluation of samples stored in

refrigerated conditions as suggested by Harry and Hildegarde

(1998)

The beverage samples (250 mL) were presented to the

trained sensory panel in capped glass jars at 5degC Samples were

kept in a cold water bath to maintain serving temperature

Samples were presented according to a random order balanced

design and room temperature dist i l led water for r insing a napkin

and score sheet on an off-white f iberglass tray Penelists

evaluated samples in standard sensory panel booths containingan

attribute definit ion sheet stop watch and pencil Panelists were

rewarded for participation after each session The coded samples

were presented to the judges in a randomized order twice a day

The evaluation performa were provided to judges for scoring as

given in appendix II

53

311 Selection of the best treatments

The functional beverages were subjected to sensory

evaluation on the basis of judges opinion based on sensory

evaluation the treatments T1 (0 β-glucan) T2 (02 β -glucan)

T3 (04 β -glucan) and T4 (06 β -glucan) were selected These

four treatments along with control (0 β -glucan) were selected for

further biological assay In control treatment pectin was used at a

concentration of 0 2 because i t is used in beverage products

very extensively

312 Efficacy studies

3121 Selection and orientation of subjects

El igibi l i ty in the program required wil l ingness and abil i ty to

adhere to the research protocol and absence of other chronic

diseases 25 healthy volunteers were selected in the program

Participation entailed both direct solicitat ion methods and

culturally tai lored efforts Direct sol ici tat ion method included

presentations face to face invitations and giving handouts that

described the study After potential participants expressed an

interest in the study they were scheduled for an orientation

Process measures included a participatory rapid appraisal a

consent form demographic questions form (including age gender

race culture income and education) and medication

questionnaire (Appendices IV) The participants were divided into

f ive groups (f ive in each) The best selected beverages were

provided to the specif ic groups in 3 replicates as mentioned in

treatment plan (Table 32) Each subject was given about 250 ml

(twice a day) of beverage every t ime

54

Table 32 Treatments used in the biological study Group Treatment (beverage)

A 0β -glucan02Pectin (Control)

B 02 β -glucan

C 04 β -glucan

D 06 β -glucan

The blood sampling of participants was carried out after

every 0 15 and 30 days of study and serum was collected through

centrifugation for analysis of different biochemical parameters in

serum

31211 Glucose level

The blood assay of the participants was carried out to

determine the blood glucose concentration Blood was taken in the

morning to determine the fasting (10-12 hrs) level of glucose and

again 1 and 2 hours after ingestion of specif ic treatment Analysis

of serum glucose was performed through Microlab-300 (Merck)

31212 Total cholesterol

The total cholesterol in the collected serum of individual

subjects of al l groups was measured by l iquid cholesterol CHODndash

PAP method as described by Stockbridge et a l (1989)

3 1213 Low density lipoprotein (LDL)

55

The low density l ipoprotein (LDL) in the serum of each

individual was measured by fol lowing the procedure of

McNamara et a l (1990)

31214 High density lipoprotein (HDL)

The serum high density l ipoprotein (HDL) was measured by

HDL cholesterol precipitant method as described by Assmann

(1979) to f ind out the impact of prepared beverages on the HDL

level of specif ied groups of participants

31215 Triglycerides (TG)

Total tr iglycerides in the collected serum of individual

participant were measured by l iquid triglycerides GPO - PAP

method as described by Annoni et a l (1982)

3 12 Statistical analysis

The data were subjected to analysis of variance (ANOVA) using

CoStat-2003 software package as described by Steel et a l (1997)

The Duncun Multiple Range (DMR) was used to determine the

level of s ignif icance between samples

56

CHAPTER- 4

RESULTS

AND

DISCUSSION

41 Chemical Composition of Barley Flour

The barley grains were cleaned and ground through Udy

cyclone sample mill and the flour was tested for different

chemical characteristics i e moisture crude fat crude protein

crude fiber ash and NFE soluble dietary fiber insoluble dietary

fiber total dietary fiber pentosans and β-glucan contents

The chemical characteristics of barley flour presented in

Table 41 indicated that the barley flour contained 1165 231

675 222 and 7707 crude protein crude fat crude fiber ash

and nitrogen free extract (NFE) respectively The results of the

present study for proximate composition of barley f lour are in line

with the earlier f indings reported for Canadian varieties by (Li et

al 2001) Helm and Francisco (2004) also concluded that Brazilian

barley varieties showed crude protein content from 1155 to

1592 crude fat 291 to 400 ash 151 to 227 and crude fiber

595 to 712 and the result of the present study fall with in the

ranges reported by these scientists Kiryluk et al (2000) have also

found crude protein content in hulled barley flour as high as

1583 and the ash content of 219 and these results also

57

Table 41 Chemical composition of barley flour

Component () on dry weight basis Crude protein 1165plusmn110

Crude fat 231plusmn021

Crude fiber 675plusmn059

Ash 222plusmn019

NFE 7707plusmn550

Soluble dietary fiber 411plusmn 039

Insoluble dietary fiber 737plusmn065

Total dietary fiber 1148plusmn109

Pentosans 303plusmn026

β-glucan 487plusmn039

58

Support to the f indings of the present study for ash content but

differed for protein content which might be due to the variation in

genetic material as well as agronomic and environmental

conditions experienced by the tested material

The results regarding chemical composit ion of barley f lour

presented in Table 41 also substantiated that barley f lour

contained higher amounts of crude f iber (675) The dietary f iber

of barley f lour in the present study was found 411 soluble

7 37 insoluble and 1148 total dietary f iber In earl ier studies

the variations in total dietary f iber soluble dietary f iber and

insoluble dietary f iber content of barley f lour have been reported

ranging from 75 to 168 56 to 64 and 19 to 104

respectively in barley (Helm and Francisco 2004 Vasanthan et a l

2002) which are very close to results found for various type of

total dietary f ibers found in the present study The results

presented in Table 41 further showed that barley f lour possessed

β -glucan 487 and pentosans 303 The results for β -glucan and

pentosans content of barley f lour in the present study are within

the ranges reported by the research workers (Papageorgiou et a l

2005 and Bhatty et a l 1991) The β -glucan is a soluble dietary

f iber component and is present in the highest amounts in the

endosperm of barley

42 Analysis of β-glucan

The β -glucan is found to be the most abundant component of the

soluble dietary f ibre in oats and barley I t is partial ly water

soluble and a l inear polysaccharide comprising only glucose units

The results regarding β -glucan given in Table 42

59

Table 42 Chemical Analysis of β-glucan

Component ()

Moisture 355plusmn029

Crude protein 996plusmn089

Crude fat 117plusmn008

Crude fiber 722plusmn055

Ash 172plusmn014

NFE 7638plusmn699

Soluble dietary fiber 7505plusmn588

Insoluble dietary fiber 1025plusmn102

Total dietary fiber 8530plusmn679

Pentosans 263plusmn019

Starch 190plusmn017

β-glucan 487plusmn039

60

indicated that β -glucan possessed 996 117 722 172 and

7638 of crude protein crude fat crude f iber ash and nitrogen

free extract (NFE) respectively

The present results regarding chemical composit ion β -glucan

are also in close agreement with the f indings reported by Bhatty

(1993) who demonstrated 33 ash content of β -glucan extracted

from barley bran The ash content (Table 42) found in the present

study is also in close conformity with the previous work of

Burkus and Temell i (2005) who reported ash content up to 4 in

β -glucan gum The pentosans contents in the present study are

also inl ine with the results reported by Burkus and Temell i (2005)

The fat content in the β -glucan was found higher as

compared to reported by Faraj et a l (2006) who found 005

lipids in high purity β -glucan concentrate which might be due to

less impurity of β -glucan extracted in the present study The

contents of starch soluble dietary f iber insoluble dietary f iber

and total dietary f iber recorded during the present study are also

in consistent with the earl ier f indings of Faraj et a l 2006) who

found variation from 04- 1 43 in starch content of β -glucan in

soluble dietary f iber (SDF) range from 7181ndash7575 and the in

insoluble dietary f iber (IDF) content of β -glucan gum pellets in

the range of (8 77-173) Symons and Brennan (2004) reported

range of 848 to 9162 for total dietary f iber (TDF) of β -glucan

which also support the results obtained for this parameter in this

present study Lambo et a l (2005) reported that barley f iber

concentrate contained 798 of total dietary f iber which is very

close to the results obtained for total dietary f iber

61

43 Analysis of β-glucan beverage

431 Color

4 3 11 L-value

The statist ical results regarding L-value measured through

colorimeter of different beverages prepared by incorporation of β -

glucan at different levels are shown in Table 43 I t is obvious

from the statist ical results that both treatments and storage

intervals exhibited signif icant effect on the L-value of different

beverages The interaction between the both the variables was

found to be non signif icant for this value of color

The color index of different beverages shown in Table 44

indicated that L-value of beverages increased as the level of β -

glucan increased in the formulation of different beverages The

results revealed signif icantly the highest L-value (2128) for

beverages of T6 containing 10 β -glucan which decreased as the

β -glucan level was reduced in the beverages and 1969 L-value

was recorded for control beverage (without β -glucan) The results

(Table 44) further showed that beverage of T5 containing 08 β -

glucan and T6 beverage containing 10 β -glucan fal l stat ist ical ly

in the same group with respect to this color values Similarly non

signif icant differences existed among beverages T2 (02 β -

glucan) T3 (04 β -glucan) and T4 (06 β -glucan) for L-value

for color

The effect of storage on the L-value of different beverages

containing different levels of β -glucan is shown in Table 44

62

Table 43 Mean sum of squares for color values (L a b) of stored β-glucan beverages

SOV df L-value a-value b-value

Treatments (T) 5 8640 48371 4088

Storage intervals (S) 6 16546 8071 17226

T x S 30 0084NS 0027NS 0964NS

Error 84 0052 0048 0164

Highly Significant (Plt001)

NS Non Significant

63

Table 44 Effect of treatments and storage intervals on the L-value of stored β-glucan beverages

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Mean

T1 (0 β-glucan) 2160 1997 1963 1950 1933 1897 1880 1969c

T2(02 β-glucan) 2213 2043 2040 1983 1973 1920 1913 2012b

T3(04 β-glucan) 2240 2073 2020 1993 1973 1950 1933 2026b

T4(06 β-glucan) 2270 2077 2030 2027 1990 1970 1940 2043b

T5(08 β-glucan) 2337 2140 2117 2120 2070 2120 1980 2126a

T6(1 β-glucan) 2263 2130 2130 2143 2080 2077 2074 2128a

Mean 2247a 2077b 2050bc 2036cd 2003de 1989ef 1953f

64

It is evident from the results that L-value of β-glucan beverages

declined significantly as a function of storage The fresh beverage

possessed the highest L-value (2257) that reduced to 2036 and

1953 when tested after 45 and 90 days of storage

It is important to note that with the increase of level of β-

glucan in the beverages affected significantly the L-value or

brightness of beverage The present study indicated that

incorporation of β-glucan resulted in improvement of beverages

color as compared to the control beverage which was prepared by

the addition of 02pectin without addition of β-glucan More L-

value by the addition of β-glucan obtained in the present study is

in consistent with the previous f indings of Bensema (2000) who

found similar pattern for increasing in L-value due to

supplementation of β-glucan However decline in L-value during

storage may be attributed to the cloud loss in the beverage

containing with β-glucan as reported by Cortes et al (2008) The

decrease in L-value was more persistent during first two weeks

but a bit stabilized after third week of storage A small amount of

precipitate was visible at the bottom of the β-glucan beverage

which is due to insoluble protein and fiber components present in

the β-glucan at low levels The precipitation of this material in case

of β-glucan supplemented beverage might be a cause of higher L-

value for these treatments of beverage as reported by Temelli et al

(2004) who prepared orange flavoured barley β-glucan beverages

and showed changes during twelve weeks storage intervals

65

4312 a-value

The analysis of variance pertaining to the a-value of

different beverages prepared by incorporation of β-glucan at

different levels indicated that both treatments and storage

intervals showed signif icant effect on the a-value of different

beverages (Table 43) However the interaction between both

variables was found non signif icantly different for a-value

The a-values of different beverages presented in Table 45

revealed that signif icantly the highest a-value (227) was

observed in beverage of T1 control beverage (without β -glucan)

while the lowest a-value (128) was possessed by T4(04 β -

glucan) I t is obvious from the results that a-value of beverages

showed upword trend as the level of β -glucan increased in the

beverage formulations This indicated decrease in the intensity of

red color in the beverages as a result of β -glucan addition in the

beverages The results further substantiated that beverages of T4

(06 β -glucan) and T6 (10 β -glucan) fal l stat ist ical ly in the

same group with respect to a color value

The results for a-value of different beverages prepared by

the incorporation of β -glucan shown in Table 45 indicated that

a-value of β -glucan beverages decreased signif icantly by

increasing the storage intervals The beverage prepared fresh got

the highest a-value (290) which declined to 144 and 099 after 45

66

Table 45 Effect of treatments and storage intervals on the a-value of stored β- glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 278 277 253 250 226 207 101 227a

T2(02 β-glucan) 267 143 120 120 113 110 107 140cd

T3(04 β-glucan) 299 155 139 130 110 099 098 147bc

T4(06 β-glucan) 280 133 127 100 090 083 083 128e

T5(08 β-glucan) 320 160 150 143 137 123 121 165b

T6(1 β-glucan) 300 130 126 118 103 085 084 135de

Means 290a 166b 153bc 144bcd 130cd 118d 099e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

67

and 90 days of storage intervals respectively A decrease in the a-

value indicated that beverage became less reddish intensity with

progress in storage periods Moreover a maximum change in the

red intensity was recorded during the f irst week of storage as

compared to the upcoming storage weeks Sa acute nchez-Moreno et a l

(2005) have reported a decl ine in a-value in pasteurize orange

juice during storage which supports to our f indings

In the present study a-value decreased signif icantly by

increasing the level of β -glucan in the beverages which indicated

that increased β -glucan concentration resulted in a less reddish

product as compared to the control beverage The results of

present study are not incormity with the f indins of Bensema

(2000) who reported increasing trend of a-value in case of β -

glucan incorporation into barley β -glucan beverage with whey

protein Isolate and found shelfstabil i ty within twelve weeks

storage at refrigeration temperature A decrease in a-value was

more persistent during f irst three weeks but a bit stabil ized after

third week

4313 b-value

The statist ical results showed that b-value of the color

index of beverages containing β -glucan at different levels was

signif icantly affected due to treatments and storage intervals

(Table 43) However the interaction between treatments and

storage intervals was found to be non signif icant for this attr ibute

of color

The beverages prepared from control treatment T1 with

02 pectin gave the highest b-value (1080) fol lowed by

68

Table 46 Effect of treatments and storage intervals on the b-value of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 1050 1077 1100 1070 1080 1080 1100 1080a

T2(02 β-glucan) 1200 960 920 920 913 900 920 962c

T3(04 β-glucan) 1240 980 960 940 930 940 900 984c

T4(06 β-glucan) 1277 1020 960 980 930 927 960 1008bc

T5(08 β-glucan) 1300 983 940 950 960 950 940 1003bc

T6(1 β-glucan) 1337 1060 1020 1007 987 997 980 1055ab

Means 1234a 1013b 983b 978b 967b 966b 967b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

69

beverage T6 (1 β -glucan) The lowest b-value was recorded in

beverage T2 (02 β -glucan) I t is obvious from the results that

incorporation of β -glucan in the beverage formulations exerted

signif icant response towards b-value of beverages when added at

1

The results in Table 46 also indicated that b-value of

different beverages decreased signif icantly as a function of

storage The freshly prepared beverages got the highest b-value

(1234) which declined to 976 after 45 days and to 967 at the

expiry of the experiment (90days) The beverages containing β -

glucan yielded more yellowish color I t is also obvious from Table

46 that decrease in b-value of beverages was more persistent

with signif icantly reduced during f irst two weeks of the storage

and beyond this period insignif icant change in b-value was

recorded up to expiry of the study i e 90 days of storage The

results of present study are in close agreement with the previous

f inding of Rodrigo et a l (2003) who showed a signif icant

decrease of b-value on pasteurized orangendashcarrot juices when

processed at 77 0C and stored at 100C stable for a period of 32

days

The addition of β -glucan at a level of 1 beverage showed

signif icant effect on b-value However b-value of different

beverages decreased as storage periods progressed This decrease

was more during the f irst two weeks of storage The decline in b-

value observed during the f irst two weeks may be due to the

precipitation of insoluble material present in the beverages or

changes in the β -glucan colorant Bensema (2000) substantiated

that b-value of beverage was reduced from 124 to 94 during the

70

refrigerated storage of 12 weeks which is in l ine with the present

results as similar reducing trend of b-value of beverages

observed in the present study The values measured as L a and

b through colorimeter represent brightness red to green and

yellow to blue color components respectively which decrease

signif icantly during the f irst two weeks of storage for al l

beverages and stabil ized later on The decrease in color values

during f irst two weeks may be attr ibuted to precipitation of

insoluble material present in beverages or change in β -carotine

colorant as reported by Temell i et al (2004) who also explained

that these precipitate are made from insoluble protein and fiber components

present in the β-glucan gum pellets at low levels during extraction procedure

432 Viscosity

The statist ical results in Table 47 showed signif icant effect

of treatments on viscosity of beverages prepared from different

concentrations of β -glucan However the storage intervals and

interaction of these two variables exhibited non signif icant effect

on viscosity of different beverages

The results in Table 48 showed that beverage prepared from

1 β -glucan incorporation (T6) possessed signif icantly the highest

viscosity (2175 mPa-s) fol lowed by T5 beverage containing (08

β -glucan) The lowest viscosity was recorded in T1 (0 β -glucan)

I t is also evident from the results in Table 48 that viscosity of

beverages increased progressively by increasing the level of β -

glucan in the formulation of beverages

I t was observed that incorporation of β -glucan showed

improvement in viscosity of beverage which might be due to the

71

Table 47 Mean sum of squares for viscosity specific gravity and total soluble solids (TSS) of stored beverages

SOV df Viscosity Specific gravity TSS

Treatments (T) 5 10026629 0003148 NS 16948375

Storage intervals (S) 6 06149915 NS 94524e-4 NS 05463508 NS

T x S 30 01087928NS 45238e-5 NS 0001213NS

Error 84 04246667 00019 03711897

Highly Significant (Plt001) NS Non Significant

72

Table 48 Effect of treatments and storage intervals on the viscosity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 441 441 439 442 447 441 450 443f

T2(02 β-glucan) 696 697 698 702 701 703 707 701e

T3(04 β-glucan) 1195 1201 1205 1218 1227 1232 1243 1217d

T4(06 β-glucan) 1607 1614 1628 1640 1651 1660 1662 1637c

T5(08 β-glucan) 1930 1935 1944 1951 1962 1968 1977 1952b

T6(1 β-glucan) 2130 2141 2152 2160 2172 2180 2287 2175a

Means 1333a 1338a 1344a 1352a 1360a 1364a 1388a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

73

presence of polysaccharides (1rarr3 1rarr4 β -glucan l inkages) The

addition of β -glucan to water also results in the formation of a

viscous hydrocolloid solution (Dawkins and Nnanna 1995

Burkus 1996) which might be one of the reasons towards increase

in the viscosity of beverages The polysaccharides hydroxyl

groups are available to form hydrogen bonds with water which

makes the polymer water-soluble Similarly Glicksman (1982) also

demonstrated that presence of the polymers in solution creates a

random network which increases the internal fr ict ion within the

solution This results in an inhibit ion to internal f low and thus

increases the viscosity of the solution by the incorporation of β -

glucan in the beverage Therefore β -glucan offers various

applications l ike beverages where other thickeners stabil izers or

gell ing agents such as pectin carrageenan guar and xanthan gum

may be replaced The results of the present study are in l ine with

the previous f indings of Bensema (2000) who observed similar

increase in viscosity of beverage by the addition of β -glucan

Thus i t may be inferred from the present results that the

thickening and stabil ization properties of barley β -glucan may be

advantageous in a beverage formulation Temell i et a l (2004)

have reported a sl ight decrease in viscosity in some beverages

containing higher hydrocolloids content (07) and found stable

viscosity in al l other beverages They also found stabil i ty of β -

glucan within the low pH in beverage formulations These

f indings support the results found in the present study

74

433 Specific gravity

The statist ical analysis pertaining to the specif ic gravity of

different beverages prepared by incorporation of β -glucan at

different levels is shown in Table 47 I t is evident from the

results that treatments storage intervals and interaction between

treatments and storage intervals showed non signif icant effect on

specif ic gravity of different beverages

The specif ic gravity of different beverages shown in Table

49 varied from 103 to 106 gL among different beverages

Mugula et a l (2001) observed sl ight decrease in specif ic gravity

in pasteurized and unpasteurize togwa samples These f indings

support the present study as non signif icant trend for this

parameter

The study of Tiisekwa et a l (2000) also showed small

variation in specif ic gravity in Tanzanian fermented beverages

when stored at ambient temperature that also supports the

present study

434 Total Soluble Solids (TSS)

The statist ical results presented in Table 47 indicated that

total soluble solids of different beverages were signif icantly

affected by treatments however storage intervals and interaction

between storage and treatments showed non signif icant effect on

TSS of different beverages

The results in Table 410 showed that the beverage

containing the highest level of β-glucan 1 (T6) possessed the

highest contents of total soluble solids (1042ordmbrix) fol lowed by

T5 beverage containing 08 β -glucan The lowest total soluble

75

Table 49 Effect of treatments and storage intervals on the specific gravity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 102 102 102 102 103 103 104 103a

T2(02 β-glucan) 102 102 103 103 103 103 104 103a

T3(04 β-glucan) 103 103 103 103 104 104 105 104a

T4(06 β-glucan) 103 104 104 105 105 106 106 105a

T5(08 β-glucan) 104 104 105 105 105 106 106 105a

T6(1 β-glucan) 105 105 105 106 106 106 106 106a

Means 103a 103a 104a 104a 104a 105a 105a Means carrying same letters within a column or row do not differ significantly (P lt 001)

76

Table 410 Effect of treatments and storage intervals on the total soluble solids of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 951 954 960 968 975 986 994 970c

T2(02 β-glucan) 950 957 960 971 980 991 1003 973c

T3(04 β-glucan) 972 977 981 988 996 1004 1013 990bc

T4(06 β-glucan) 989 992 995 1006 1016 1026 1037 1009abc

T5(08 β-glucan) 1001 1005 1009 1017 1027 1039 1048 1021ab

T6(1 β-glucan) 1019 1026 1031 1042 1052 1060 1067 1042a

Means 980a 985a 989a 999a 1008a 1018a 1027a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

77

solids (970ordmbrix) were yielded by the beverage of T1 (0 β -

glucan) I t is obvious from the results that total soluble solids of

beverages increased progressively by increasing the level of β -

glucan in beverage formulations

The total soluble sol ids in different beverage did not differ

signif icantly as a function of storage The total soluble solids in

the freshly prepared β -glucan beverages were found 980 ordmbrix

and total soluble solids 1027ordmbrix were recorded in the beverages

tested of the experiment (day 90) The present study is supported

by the f indings of Mugula et a l (2001) who explained that TSS

decreased in unpasteurized and pasteurized beverage prepared

from sorghum The f indings of present study are also in l ine with

the observations of Tiisekwa et a l (2000) In other study Akubor

(2003) also repoted similar results in melon-banana beverage

during ambient temperature storage

435 pH

The results regarding pH of different β -glucan supplemented

beverages presented in Table 411showed that pH of the

beverages was not affected by the treatments and interaction

between treatments and storage intervals The pH of different

beverage was signif icantly affected by the storage intervals

The results regarding pH of the beverages given in Table 412

indicated non signif icant changes in pH due to different levels of

β -glucan supplementation

78

Table 411 Mean sum of squares for pH acidity and ascorbic acid content of stored β-glucan beverages

SOV df pH Acidity Ascorbic acid

Treatments (T) 5 0014 0084 111646

Storage intervals (S) 6 0227 0008 2447942

T x S 30 0001NS 00001NS 13116NS

Error 84 0004 00002 30928

Highly Significant (Plt001) NS Non Significant Significant (Plt001)

79

Table 412 Effect of treatments and storage intervals on the pH of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 301 294 288 284 279 276 271 285a

T2(02 β-glucan) 297 291 285 280 274 271 268 281a

T3(04 β-glucan) 303 298 292 288 281 274 273 287a

T4(06 β-glucan) 303 296 293 287 283 276 274 287a

T5(08 β-glucan) 296 292 288 281 277 273 269 282a

T6(1 β-glucan) 305 301 288 284 281 273 265 285a

Means 301a 295ab 289bc 284cd 279cde 274de 270e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

80

The results in Table 412 showed a signif icant effect of storage

intervals on the pH value of different beverages The pH value of

freshly prepared beverages (0 day) was found signif icantly higher

301 which decreased to 270 when beverages tested after (90

days) The pH values decreased signif icantly in al l the beverages

progressively throughout the storage period The results of the

present study with respect to storage studies are in concordance

with the f indings of (Miguel et a l 2004 and Falade et a l 2003) who

found a decreasing trend of pH in beverages during storage Ziena

(2000) reported a gradual decline in pH and showed a percent

decrease in pH values range from 11 to 87 in refrigerated and

freeze l ime juices samples High acid and low pH may be due to

production of acetic acid and lactic acid during storage Such

types of changes in pH vales have been demonstrated by (Souci et

a l 1987 Kaanane et a l 1988 Martin et a l 1995) The results are

in consistent with the f indings of Akubor (2003) who also

reported drop in pH with storage period in melon-banana

beverage

Fasoyiro et a l (2005) have founded a decrease in pH during

storage at 50C The Roselle beverage containing three different

fruits (orange apple and pineapple) was prepared They found

decrease in pH from 354 to 280 during two weeks storage at

refrigeration temperature The reduction in pH may be due to the

decomposit ion of fermentable polysaccharides i e β -glucan

sucrose and high fructose corn syrup which are present in

beverages This sl ight decrease in pH is a function of refrigeration

temperature storage which slows down the rate of growth of

microorganisms during entire period of cold storage

81

436 Acidity

The statistical results regarding acidity of beverages

prepared from different levels of β-glucan presented in Table 411

indicated that acidity of beverages was significantly affected by the

storage intervals however treatments and interaction between

storage treatments showed non significant effect on the acidity of

different beverages

The results in Table 413 further substantiated a non

significant effect due to different levels of β-glucan for different

beverages The acidity of different beverages differed significantly

which was found 160 in the fresh beverages The acidity was

increase linearly as the storage progressed which reaches 161 at

the end of experiment (three months) during storage period

Alessandra et al (2004) also reported similar results which

supports the present findings for increase in acidity during

storage The acidity increased significantly as a function of storage

of orange juice stored at 4 0C (137 g100g) and at 10 0C

(136g100g) after 4 and 3 weeks of storage respectively (Esteve et

al 2005)

During two weeks change in acidity was recorded from

190 to 225 in Roselle orange drink (Fasoyiro et al 2005) which

also supports the results of present study The gradual increase in

acidity was due to refrigeration temperature The decrease in pH

and increase in acidity during storage might be due to degradation

of sucrose high fructose corn syrup and β-glucan by the action of

microorganisms which causes production of acids in beverages

82

Table 413 Effect of treatments and storage intervals on the acidity of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 140 146 147 150 152 154 160 150a

T2(02 β-glucan) 139 144 144 147 153 156 157 149a

T3(04 β-glucan) 144 146 153 154 156 159 162 153a

T4(06 β-glucan) 143 145 153 151 155 160 163 153a

T5(08 β-glucan) 141 144 148 153 156 161 160 152a

T6(1 β-glucan) 144 145 150 154 158 160 162 153a

Means 142a 145b 149c 152d 155e 158f 161g

Means carrying same letters within a column or row do not differ significantly (P lt 001)

83

437 Ascorbic acid

The results regarding analysis of variance for ascorbic acid

content of different beverages prepared from different levels of β -

glucan have been presented in Table 411 The statist ical results

indicated that ascorbic acid content of different beverages was

affected signif icantly due to storage intervals but differed non

signif icantly due to treatments and interaction between

treatments and storage intervals

The results in Table 412 showed non signif icant change in

ascorbic acid content due to incorporation of β -glucan

The ascorbic acid content was found higher a (29406 mgkg)

in fresh beverage which declined signif icantly to 27933 mgkg

and 26211 mgkg after 45 and 90 days storage of beverages

respectively I t is also evident from results that ascorbic acid

content of beverages decreased consistently as storage period

increased

The f indings of the present study is in l ine with the work

reported by different researchers Crandall et a l (1987) and Maria

et a l (2003) who observed a signif icant loss of ascorbic acid (25 to

26) during storage In the present study the ascorbic acid

content decreased with the increase in storage periods This

decrease might be due to the factors such as storage temperature

oxidative enzymes processing techniques metal contamination

and the presence of atmospheric oxygen in the head space

Kabasakalis et a l (2000) studied the ascorbic acid content of

commercial fruit juices and observed that the loss of ascorbic acid

84

Table 414 Effect of treatments and storage intervals on the ascorbic acid contents of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 29333 29033 28333 28067 27667 27067 26400 27986

T2(02 β-glucan) 29733 29133 28300 27900 27133 26333 25767 27757

T3(04 β-glucan) 29167 28733 28600 28100 27133 26767 26100 27800

T4(06 β-glucan) 29300 28867 28267 27367 27167 26400 25900 27610

T5(08 β-glucan) 29600 29400 28967 28300 27500 27300 26867 28276

T6(1 β-glucan) 29300 28767 28300 27867 27400 26900 26233 27824

Means 29406a 28989ab 28461bc 27933cd 27333de 26794ef 26211f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

85

was 29-41 in commercial fruit juices stored in closed container

at room temperature for 4 months Similar results reported by

Otta (1984) who described gradual decrease in ascorbic acid at

refrigeration temperature due to prolong storage Since in the

present study the beverages were stored at refrigeration

temperature therefore the loss in ascorbic acid is in conformity

with the results of Otta (1984)

86

438 Reducing Sugars

The statistical results regarding reducing sugars of beverages

presented in Table 415 indicated that the reducing sugars of

beverages were affected significantly by the storage intervals

However the treatments and the interaction between treatments

and storage intervals showed non significant effect on the reducing

sugars of different beverages

The results for the reducing sugars of beverages prepared

from different treatments of β-glucan are presented in Table 416

which indicated that reducing sugars of beverages did not differed

significantly due to the incorporation of β-glucan in different

beverages

The reducing sugars it increased significantly from 372 to

431 during 0 to 90 days of storage respectively (Table 416) In

fresh beverage samples the reducing sugar content was found 372

mg which increased to 402 and 431 mg after 45 and 90 days of

storage respectively The results showed that reducing sugar

contents of beverage increased slowly in the first 15 days of

storage but increased consistently and rapidly as the storage

period increased indicating more production of reducing sugars in

the beverage samples in the later stages of storage periods

Babsky et al (1986) studied storage effect on the composition

of clarif ied apple juice concentrate and reported that reducing

sugars increased from 0286 to 0329 moles per 100 grams and

sucrose decreased from 0039 to 0015 moles per 100 grams after

111 days of storage The reducing sugars were formed by the

inversion of sucrose hydrolysis effect of temperature as described

87

Table 415 Mean sum of squares for reducing non reducing and total sugar content of stored β-glucan beverages

SOV df Reducing Sugars Non Reducing Sugars Total sugars

Treatments (T) 5 00092NS 0004NS 00087265NS

Storage intervals (S) 6 0837 0357 01086119 NS

T x S 30 0001NS 0001NS 8954e-4 NS

Error 84 0003 0004 01528365

Highly Significant (Plt001) NS Non Significant

88

Table 416 Effect of treatments and storage intervals on the reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 371 380 391 402 412 421 431 401

T2(02 β-glucan) 373 383 390 400 409 419 427 400

T3(04 β-glucan) 371 379 389 402 413 421 434 401

T4(06 β-glucan) 368 380 392 402 414 424 432 402

T5(08 β-glucan) 375 382 394 408 417 427 435 405

T6(1 β-glucan) 372 382 389 400 409 417 427 399

Means 372f 381ef 391de 402cd 412bc 422ab 431a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

89

by Ranote and Bains (1982) and Stein et al (1986) Increases in

total sugars have also been observed by Godara and Pareek (1985)

in date palm juice during storage at room temperature

The increase in reducing sugars have also been reported by a

number of research workers and the reason shown to increase in

this parameter has been due to conversion of non reducing sugars

to reducing sugars with the increased storage duration as reported

by Purthi et al (1984) He also reported an increase in reducing

sugars from 136 to 238 per cent and a decrease in non-reducing

sugars from 296 to 230 per cent at room temperature during

storage in juices of four commercial varieties of malta and orange

The results are in close confirmatory with the finding of (Fuleki et

al 1994) who also reported increases in fructose from 412 to 676

and glucose from 070 to 227 in fruit juices during storage

439 Non Reducing Sugars

Non reducing sugars of beverages stored for a period of

three months was not affected significantly by the treatments

(Table 415) The storage intervals showed significantly effect on

non reducing sugars of different beverages The interaction

between treatments and storage intervals possessed non significant

effect on non reducing sugars of different beverages

The contents of non reducing sugars of different beverages

were not significantly changed due to incorporation of different

levels of β-glucan

The results in Table 417 revealed that non reducing sugars

decreased significantly as a function of storage The non reducing

sugars were found significantly the highest content (514) in fresh

90

Table 417 Effect of treatments and storage intervals on the non reducing sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 514 508 501 493 487 481 476 494a

T2(02 β-glucan) 515 509 504 497 490 483 478 497a

T3(04 β-glucan) 513 507 501 494 487 482 475 494a

T4(06 β-glucan) 517 511 503 496 490 482 477 497a

T5(08 β-glucan) 512 507 501 493 486 480 474 493a

T6(1 β-glucan) 513 506 502 493 486 481 476 494a

Means 514a 508ab 502bc 495cd 488de 482ef 476f

Means carrying same letters within a column or row do not differ significantly (P lt 001)

91

beverages which reduced to 495 and 476 after 45 and 90 days of

storage respectively

The f indings of the present study are well supported by

Singh et a l (2007) who found that with increase in storage t ime

non-reducing sugars decreased The results are also in l ine with

the f indings of Chowdhury et a l (2008) who studied the six

months storage effect on the shelf l i fe of mixed juice and

signif icant decrease in non reducing sugars due to breakdown of

non reducing sugars (sucrose) with the reaction of acids

4310 Total Sugars

The analysis of variance regarding total sugars of beverages

showed that total sugars were non signif icantly affected due to

treatments and storage intervals as well as the interaction

between treatments and storage intervals (Table 415)

The results for total sugars of different beverages

presented in Table 418 substantiated that the total sugars content

in al l the treatments fel l stat ist ical ly the same group and total

sugars remained unchanged by the incorporat ion of β -glucan in

the beverages The total sugar content of β -glucan supplemented

beverages s tored for a period of 3 months indicated a lso showed

non s ignif icant var iat ion between the freshly prepared β -g lucan

beverages and beverages evaluated af ter 90 days of s torage

studies The results are wel l in agreement with the observations

92

Table 418 Effect of treatments and storage intervals on the total sugars of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 885 888 892 895 899 902 907 895a

T2(02 β-glucan) 888 892 894 897 899 902 905 897a

T3(04 β-glucan) 884 886 890 896 900 903 909 895a

T4(06 β-glucan) 885 891 895 898 904 906 909 898a

T5(08 β-glucan) 887 889 895 901 903 907 909 899a

T6(1 β-glucan) 885 888 891 893 895 898 903 893a

Means 886a 889a 893a 897a 900a 903a 907a

Means carrying same letters within a column or row do not differ significantly (P lt 001)

93

of Chowdhury et a l (2008) who reported non signif icant increase

in total sugars up to six months storage at 28 0C in juices

4 4 Total Plate Count (TPC) of the beverage samples

The results in Table 419 indicated that storage intervals

showed decline in total plate count (TPC) of β -glucan beverage

The TPC value of freshly prepared beverage (0 day) was higher

129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times 104 at

the end of the experimental study (90 day) Similar counts of TPC

have been reported for some juices and drinks in Egypt (Daw et a l

1994) These results are also in agreement with those of Hancioglu

amp Karapiner (1997) reported for Turkish boza beverages The

contamination by these microorganisms in the beverages could

have occurred during processing and packaging as most of the

people involved in the production and packaging do not take

necessary precautions Contamination of food items may largely

be due to the presence of these organisms and their entrance into

the food or beverage as a result of poor hygiene and sanitation

conditions (Bibek 2001)

The results indicated that the TPC values decreased in al l

the beverages containing throughout the storage period The

results of the present study with respect to storage period are in

consistent with the f indings of other researchers who reported

similar results for some tradit ional beverages and drinks (Daw et

a l 1994) The TPC values decrease gradually during storage

intervals are this might be due to

94

Table 419 Effect of treatments and storage intervals on the total plate count (CFUml) of stored β-glucan beverages

Storage intervals (days) Treatments

0 15 30 45 60 75 90

T1 (0 β-glucan) 187 x 104 187 x 104 184 x 104 179 x 104 172 x 104 169 x 104 166 x 104

T2(02 β-glucan) 252 x 104 247 x 104 247x 104 239 x 104 239 x 104 233 x 104 233 x 104

T3(04 β-glucan) 366 x 104 363 x 104 360 x 104 357 x 104 357 x 104 352 x 104 348 x 104

T4(06 β-glucan) 318 x 104 316 x 104 315 x 104 315 x 104 312 x 104 310 x 104 308 x 104

T5(08 β-glucan) 446 x 104 443 x 104 442 x 104 441 x 104 439 x 104 439 x 104 432 x 104

T6(1 β-glucan) 129 x 104 129 x 104 125 x 104 123 x 104 119 x 104 119 x 104 117 x 104

95

increase in acidity which may cause a concomitant decrease in pH

value which may help to decrease TPC in the beverages (Kaanane

et a l 1988 Martin et a l 1995) The total bacterial counts obtained

in this study fal l between 10 x 102 - 1 0 x 105 CFUml which fal l

within the range of earl ier works done by Hatcher et a l (1992)

45 Sensory evaluation of β -glucan beverages

451 Color

The analysis of variance pertaining to the color scores

assigned to different treatments of beverages by the panelist

indicated that color of beverages differed signif icantly due to the

treatments and storage intervals (Table 420) However the

interaction between treatment and storage intervals showed non

signif icant effect on this sensory attribute

The scores assigned to the color of different beverages

prepared by incorporation of β -glucan presented in Table 421

revealed that the beverage prepared by the incorporation of 0 2

β -glucan got signif icantly the highest color scores (684) fol lowed

by the control beverage (02 pectin) The panelists assigned the

lowest scores (494) to the color of T6 beverage (10 β -glucan) I t

is evident from the results (Table 421) that the beverages of

treatments T1 (control) T2 (02 β -glucan) T3 (04 β -glucan)

and T4 (06 β -glucan) fel l stat ist ical ly in the same group with

respect to color scores The results also indicated non signif icant

differences in color scores between beverages T5 (08 β -glucan)

and T6 (10 β -glucan) The beverages containing β -glucan level

up to 06 remained acceptable by the panelists however further

96

Table 420 Mean sum of squares for sensory evaluation of stored β-glucan beverages

SOV df Color Flavor Sweetness Sourness Overall acceptability

Treatments (T) 5 24686 18760 18873 9970 34811

Storage intervals (S) 6 13933 27297 59231 22338 62242

T x S 30 0526NS 0283NS 0169NS 0987NS 0125NS

Error 108 0436 0383 0388 1936 0626

Highly Significant (Plt001)

NS Non Significant

97

Table 421 Effect of treatments and storage intervals on the color score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 68 68 60 52 663a

T2(02 β-glucan) 80 74 72 68 66 62 56 683a

T3(04 β-glucan) 78 72 70 70 68 54 48 657a

T4(06 β-glucan) 72 66 64 60 56 54 50 603a

T5(08 β-glucan) 58 52 50 46 50 48 46 500b

T6(1 β-glucan) 54 54 52 50 48 46 42 494b

Means 693a 650ab 630ab 603bc 593bc 540cd 490d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

98

increase in the β -glucan level in beverages resulted decrease in

assigning scores to color I t is obvious that freshly prepared β -

glucan beverage got maximum scores for color (693) which

reduced to 490 scores when evaluated at the end of the

experiment (90 days) The results showed that the panelists l iked

more the color of fresh beverages and this l iking reduced of

beverages stored (Table 421)

Colour of any food product is an important criterion for the

acceptabil i ty of any food product I t is one of the characterist ics

perceived by the senses and a mean for the rapid identif ication

and ult imately governs the acceptance or re jection of the food

product The results obtained in the present study for color score

are in l ine with the f indings of Anjum et a l (2006) who observed

signif icant effect (p lt 0001) on color parameters during different

storage conditions Thus the beverages of different treatments got

signif icant variation in gett ing score for their color yet the score

assigned to the color after 90 days under refrigerated storage

remained acceptable The change in color parameter may be due to

the mail lard reaction between reducing sugars and amino acids

(Gonzalez amp Leeson 2000) The results are in close agreement

with the f indings of Granzer (1982) who also reported similar

results for color of beverages at different storage periods

99

452 Flavor

The statist ical results for the scores assigned to f lavor of

beverages prepared from different β -glucan levels indicated that

f lavor score varied signif icantly due to differences (β -glucan

levels) in treatments as well as storage intervals (Table 420) The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to f lavor of different

beverage

The panelists assigned the signif icantly highest scores to the

f lavour of beverages containing 04 β -glucan (T3) (Table 422)

However the beverage treatment T6 (10 β -glucan) was ranked

at the bottom for f lavor scores (586) by the panelists The

beverages containing 06 β -glucan and control (T1) got

statist ical ly similar scores for f lavour The beverages containing

more than 06 β -glucan got lower scores for f lavor

The effect of storage on the f lavor of beverages stored for a

period of three months showed that there was signif icant decrease

in assigning the scores to the f lavour beverages as a function of

storage The fresh beverages got signif icantly the highest scores

(833) while the beverages tested after 90 days storage got the

lowest score (510) by the panelists I t is evident from the results

(Table 422) that scores assigned to f lavor of beverages decreased

as storage progressed three months

A decrease in the scores assigned to f lavor of different

beverages may be attr ibuted to the increase in acidity of beverage

which noticed during storage as reported in the earl ier section

This increase in acidity may enhance the sourness and wil l

100

Table 422 Effect of treatments and storage intervals on the flavor score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 86 82 76 74 74 62 56 729ab

T2 86 84 78 74 72 66 56 737ab

T3 92 86 80 72 74 64 60 754a

T4 80 76 68 62 64 60 52 660bc

T5 70 68 64 58 58 56 46 600c

T6 72 66 60 54 56 52 50 586c

Means 810a 770ab 710bc 657cd 663cd 600de 533e

Means carrying same letters within a column or row do not differ significantly (P lt 001)

101

depress the f lavor of beverage with the passage of t ime during

storage

A gradual decrease in f lavor during storage may also be due

to degradation of f lavour due to storage of product at refrigerator

temperature and due to heat treatment applied during processing

and such reasons for decrease in f lavor have been reported by

Pruthi et a l (1981) Hassan (1976) The change in f lavour as a

function of storage may be due to the degradation of ascorbic acid

and furfural production (Shimoda amp Osaj ima 1981 Perez amp Sanz

2001)

The productrsquos physico-chemical changes may alter f lavor

during storage The present study is well supported by the results

of Anjum et a l (2004) who described that effect of process heat

treatment and storage temperature are well correlated with the

production of off f lavoring compounds due to browning reaction

and furfural production

453 Sweetness

The scores assigned to sweetness of different beverages

differed signif icantly among treatments and storage intervals

(Table 420) However the interaction between treatments and

storage intervals showed non signif icant effect on this sensory

attr ibute

The scores assigned to sweetness of different beverages in

Table 423 revealed that the control beverage containing 02

pectin got the highest scores for sweetness (674) fol lowed the

beverage 02 β -glucan The beveraged of T6 containing 10 β -

102

glucan got the lowest scores (503) for sweetness The beverage T1

(control) and T2 (02 β -glucan) were place statist ical ly at same

level for scores given to sweetness Non signif icant differences

existed for sweetness score between beverages of T5 (08 β -

glucan) and T6 (10 β -glucan) The results also demonstrated

that the beverages containing β -glucan up to 06 got acceptable

scores however further increase in addition of β -glucan levels in

the beverages got lower scores by the panelists

The results also indicated that fresh beverages got higher

scores (700) which were reduced to 570 scores when evaluated

after 45 days of storage and to 507 scores tested after 90 days of

storage The results of the present study showed that as the

storage t ime increase the sweetness score decreasedThese

observations are well supported by the f indings of Esteve et a l

(2005) and Fasoyiro et a l (2005) who found that during storage

period pH decreases and acidity increases of juices and drinks

due to the degradation of carbohydrates by the action of

microorganisms

103

Table 423 Effect of treatments and storage intervals on the sweetness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 80 78 72 66 60 60 56 674a

T2(02 β-glucan) 80 74 70 68 60 58 58 669a

T3(04 β-glucan) 72 70 62 58 56 60 54 617ab

T4(06 β-glucan) 68 66 60 54 56 58 50 589b

T5(08 β-glucan) 58 56 50 46 50 52 46 511c

T6(1 β-glucan) 62 56 54 50 50 40 40 503c

Means 700a 667ab 613bc 570cd 553cd 547cd 507d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

104

454 Sourness

The statist ical results for the scores given to sourness of

beverages prepared by different levels of β -glucan (Table 420)

indicated that sourness scores varied signif icantly due to

differences in treatments as well as storage intervals The

interaction between treatments and storage intervals showed non

signif icant effect on the scores given to sourness of different

beverages

The scores assigned to the sourness of different beverages

given in Table 424 revealed that the highest scores (643) were

given to beverages of control treatment (T1) fol lowed by beverage

of T2 (02 β -glucan) but non signif icant differences existed

between these two beverages The beverage of treatment T6 (10

β -glucan) got the lowest scores (511) for sourness The beverage

containing 06 β -glucan and control beverage got statist ical ly

similar scores The incorporation of β -glucan more than 06

showed a declining trend in gett ing the scores for the sourness

The fresh beverages got the highest scores (697) for

sourness while the beverages tested at the expiry of study i e 90

days of storage got the s ignif icantly lowest scores for sourness

(460) I t is evident from the results (Table 424) that scores given

to sourness of beverages decreased l inearly throughout the

storage period of three months

The present study indicated that control beverage was

sl ightly sourer than the beverages containing different level of β -

glucan but the differences in scores (pectin) of sourness were not

105

Table 424 Effect of treatments and storage intervals on the sourness score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 74 72 70 66 64 56 48 643a

T2(02 β-glucan) 72 70 70 66 64 56 50 640a

T3(04 β-glucan) 76 72 72 68 62 50 46 637a

T4(06 β-glucan) 70 68 68 64 60 54 46 614a

T5(08 β-glucan) 64 62 58 56 50 50 46 551b

T6(1 β-glucan) 62 58 56 52 40 50 40 511b

Means 697a 670a 657a 620ab 567ab 527ab 460b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

106

s ignif icant with beverages containing up to 06 β-glucan This

indicated that β -glucan does not contribute to beverage sourness

intensity However there was a sl ight decl ine in sourness

intensity in the beverage with β -glucan beyond 06 Bensema

(2000) who also observed that addition of β -glucan may contribute

towards sl ight alkaline environment which reduces the sourness

The results of the present study are also in agreement with the

f indings of Pangborn et a l (1973) who showed that sourness

declined by increasing the hydrocolloid concentration in the

beverages The sensory evaluation of beverages regarding

sourness with storage got lower scores The decrease in pH may

cause increase in acidity as a function of storage which made the

beverage sourer The results obtained from the present study are

in l ine with the f indings of Fasoyiro et a l (2005) and Akubor

(2003) who recorded sl ight increase in acidity during refrigeration

storage of Roselle orange drink An increase in acidity resulted in

sourness in beverages

455 Overall Acceptability

The statist ical results for the score given to overall

acceptabil i ty of beverages (Table 420) indicated that treatments

and storage intervals s ignif icantly affected the overall

acceptabil i ty scores The interaction between treatments and

storage intervals were found non signif icant for overall

acceptabil i ty scores

The beverage prepared from the control treatment (T2) got

the highest overall acceptibi l i ty scores (731) fol lowed by

107

beverage of T1 (02 pectin) but both these beverages possessed

non signif icant differences for overall acceptibi l i ty scores The

beverages of T3 (04 β -glucan) and T4 (06 β -glucan) treatments

got statist ical ly overall acceptabil i ty scores The beverages of

treatments T5 (08 β -glucan) and T6 (1 β -glucan) got the lowest

scores (511) by the panelists for overall acceptabil i ty scores I t is

obvious from the results (Table 425) that overall acceptabil i ty

scores got by beverages containing up to 06 β -glucan

incorporation and control got stat ist ical ly similar scores The

beverages containing more than 06 β -glucan got lower scores

for overall acceptabil i ty

The scores for overall acceptabil i ty of beverages decreased

during storage The fresh beverages got the highest scores (737)

while the beverages tested after 90 days of storage got the lowest

overall acceptabil i ty scores

The β -glucan has been found to be stable within the acidic

environment of an orange-flavored beverage during processing

and refrigerated storage β -glucans abil i ty to increase viscosity

upon addition to water makes i t an excellent thickener for

beverage applications These characterist ics provided more appeal

to the panelists for making the decision about the overall

acceptabil i ty of beverages The results of the present study are in

l ine with the f indings of Renuka et a l (2009) who prepared fruit

juice beverages with fort i f ied fructo-oligosaccharide and noted

the quality characterist ics with six months storage period There

was negligible change in overall quality that ranges from 90 to

60 for different beverages at refrigeration temperature with

references to hedonic scale evaluation

108

Table 425 Effect of treatments and storage intervals on the overall acceptability score of stored β-glucan beverages

Storage Intervals (days) Treatments

0 15 30 45 60 75 90 Means

T1 (0 β-glucan) 84 84 80 72 72 62 54 726a

T2(02 β-glucan) 82 82 76 74 72 66 60 731a

T3(04 β-glucan) 80 80 74 70 70 62 54 700a

T4(06 β-glucan) 72 72 68 66 64 58 50 643a

T5(08 β-glucan) 62 62 60 54 54 44 40 537b

T6(1 β-glucan) 62 62 60 56 50 44 42 537b

Means 737a 737a 697ab 653abc 637bc 560cd 500d

Means carrying same letters within a column or row do not differ significantly (P lt 001)

109

Selection of best treatments

After sensory evaluation best treatments were selected for

further studies The beverages containing different levels of β -

glucan gett ing maximum scores by the judges during entire

storage period were selected Three best beverages were selected

for eff icacy study containing 02 0 4 and 06 β -glucan levels

along with control beverage containing 02 pectin as i t is

commonly used in beverages preparation

46 Efficacy studies of β -glucan beverages

461 Total cholesterol

The statist ical results regarding total serum cholesterol of

healthy subjects fed with various levels of β -glucan supplemented

beverages are presented in Table 426 The results indicated that

total serum cholesterol was signif icantly affected due to variation

in beverage formulations and study periods The interaction

between these both variables was found non signif icant for total

serum cholesterol

I t is obvious from the results given in Table 427 and

i l lustrated in Figure 41 that the highest concentration of total

cholesterol (13953 mgdl) was observed in the control group

which was fed on beverage prepared without any addition of β -

glucan The subject group fed on beverage containing 06 β -

glucan (D) possessed the lowest content of total cholesterol

(13230 mgdl) in serum of healthy subjects at the end of study I t

is evident from Figure 41 that there was signif icant and

progressive decline in the total serum cholesterol by increasing

110

Table 426 Mean sum of squares for blood lipid profile of volunteers

SOV df Total Cholesterol Triglycerides LDL HDL

Beverages (B) 3 107368 37570 55266 28197

Study Periods (S) 2 422014 398238 212944 63649

B x S 6 30566 12210 15847 7837

Error 24 0069 0031 0010 0012

Highly Significant (Plt001) NS Non Significant

111

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week3

Study Period

ABCD

210297

673

826

145

276

517456

0123456789

Decrease

Week2 Week4

Study Period

ABCD

Table 427 Effect of β-glucan supplemented beverage on serum total cholesterol

content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 14220 13921 13719 13953a

B 14174 13753 13374 13767b

C 14198 13242 12557 13332c

D 14211 13037 12442 13230d

Means 14201a 13488b 13023c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 41 decrease in the serum total cholesterol level of subjects fed on

different beverages A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

112

137191337513246

12557

1422013921

14178

13757

141951421

12442

13035

115

120

125

130

135

140

145

Base Line Week-2 Week-4

Weeks

Tota

l Cho

lest

erol

(mg

dl)

A B C D

Figure 42 Effect of β-glucan beverage on Total Cholesterol (mgdl) content of

healthy volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

113

the level of β-glucan in the beverage formulations There was a

decrease in total cholesterol content when the subjects were fed on

beverages C (04 β-glucan) and D (06 β-glucan) The results in

Figure 42 also showed that total cholesterol of healthy subjects

decreased at a faster rate during first two weeks as compared to the

last two weeks of the experimental study The highest decrease in

total cholesterol (826) content was observed in the group of

subjects fed on 06 β-glucan supplemented beverage (D) followed

by the group fed on beverage C (04 β-glucan) and the lowest

decrease in the serum cholesterol was observed in the group fed on

control beverage (0 β-glucan) both when tested at week 2 and

week 4 However Figure 42 also depicted that maximum decrease

in total cholesterol content was shown by the beverage C (04 β-

glucan) when subjects were tested after four weeks

A significant decrease in the total serum cholesterol of test

subjects was found in the present study which might be due to

different factors including the presence of β-glucan soluble dietary

fiber and tocopherol content of barley β-glucan supplemented in

beverage It is well documented that β-glucan has the ability to

reduce the blood serum total cholesterol content of different

subjects (Uusitupa et al 1992) β-glucan is a soluble dietary fiber

portion of barley and possess the ability to decrease the total

cholesterol Ornish et al (1998) have shown reduction in plasma

cholesterol concentrations due to contents of dietary fiber Brown et

al (1999) also reported that 1g of soluble fiber can lower total

cholesterol by about 0045mmolL It has been recommended by

FDA that at least 3 gday of β-glucan from barley should be

consumed to achieve a clinically relevant reduction in serum total

114

cholesterol concentrations (FDA 1996) Soluble dietary fibers may

increase the binding of bile acids in the intestinal lumen which

leads to a decreased enterohepatic circulation of bile acids and a

subsequent increase in the hepatic conversion of cholesterol to bile

acids (Bell et al 1999) Another suggested mechanism is that the

increased viscosity of the food mass in the small intestine because of

soluble fibers leads to the formation of a thick unstirred water layer

adjacent to the mucosa This layer may act as a physical barrier to

reduce the absorption of nutrients and bile acids (Beer et al 1995)

Thus these properties of β-glucan have shown a significant decline

in total cholesterol due to intake of different beverages containing

different levels of β-glucan

462 Triglycerides

The analysis of variance showed significant effect of

functional beverages and study periods on triglyceride content of

adult subjects (Table 426) The interaction between functional

beverages and study periods was found non significant for this

biochemical parameter

The results i l lustrated in Figure 44 and Table 428 indicated

the functional beverages showed different response towards level

of serum triglycerides in different adult groups I t is evident from

Figure 44 that level of serum triglyceride was higher in the

subject group fed on control beverage (0 β -glucan) while the

level of tr iglyceride content was recorded maximum in the group

fed on beverage D (06 β -glucan)It is also obvious from Figure

43 that

115

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

369 447

10431099

497

672767 757

0

2

4

6

8

10

12

Decrease

Week2 Week4

Study Period

ABCD

Table 428 Effect of β-glucan supplemented beverage on serum Triglycerides content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 8668 8348 7933 8316a

B 8547 8165 7616 8109b

C 8747 7835 7234 7939c

D 8611 7665 7085 7854d

Means 8643a 8028b 7492c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 43 decrease in the serum triglycerides level of subjects fed on different

beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

116

79337616

7234

8668

83488547

81657835

87478611

7765

7185

60

65

70

75

80

85

90

Base Line Week-2 Week-4

Weeks

Trig

lyce

ride

s (m

gdl

)

A B C D

Figure 44 Effect of β-glucan beverage on Triglyceride (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

117

reduction in the tr iglyceride contents increased by increasing the

level of β -glucan in different the beverages

The tr iglyceride content of subjects fed on different

functional beverages decreased at higher rate during the

beginning of days of the experiment There was maximum

decrease in tr iglyceride content (1099) in subject group fed on

the beverage D (06 β -glucan) when tested after week-2 while

the lowest decrease in tr iglycerides was recorded in the group fed

on beverage A (control) The rate of reduction in tr iglyceride

content was at a lower rate after 2 weeks of storage study The

beverage C (04 β -glucan) showed more pronounced effect on the

content of tr iglycerides during the last fortnight of the experiment

as compared to al l other beverages

The results regarding triglyceride contents presented in Table

428 indicated the tr iglyceride content of healthy subjects differed

signif icantly as a function of storage

The results of the present study are in agreement with the

f indings of Delaney et a l (2003a) who found a decrease in serum

triglyceride content of rats as compared to control by

administration of β -glucan in the feed The study demonstrated

that tr iglyceride content reduced progressively as the level of β -

glucan increased in the beverage and the highest reduction was

achieved by the supplementation of 0 6 β -glucan in the beverage

formulation The decrease in tr iglyceride content may be

attributed to the level of β -glucan content has the abil i ty to

reduce tr iglyceride content

118

I t is evident from the previous studies that the level of

tr iglyceride content reduced by the β -glucan incorporation in

different food products Biorklund et a l (2005) observed changes

in serum lipids and reported a total reduction of 0 14mmoll with

a diet containing 5g β -glucan from oat for a period of f ive weeks

study Similar decrease in tr iglycerides has been reported

observed by Naumann et a l (2006) who incorporated β -glucan in

to fruit drink and found a total 1 26 decrease in subjects of β -

glucan group for a period of f ives weeks I t may be concluded

from the present study that by intake of β -glucan in beverage

formulation can help to reduce the tr iglycerides content in human

subjects to a signif icant level

463 Low Density Lipoproteins (LDL)

The statist ical results regarding LDL content of adult subjects

fed on beverages supplemented with various levels of β -glucan

are shown in Table 426 The results indicated that LDL was

affected signif icantly by the variation in beverage formulations as

well as study periods The interaction between beverages and

study periods was found to be non signif icant for LDL content of

different subjects

The highest concentration of LDL (5202 mgdl) was

recorded in the subject group fed on beverage (control) without

addition of β -glucan (Table 429 and Fig 4 6) The subject group

fed on

119

433

754

14871657

111

419

769 743

02468

1012141618

Decrease

Week2 Week4

Study Period

ABCD

Table 429 Effect of β-glucan supplemented beverage on serum LDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 5376 5143 5086 5202a

B 5345 4942 4735 5007b

C 5365 4567 4216 4716c

D 5388 4495 4161 4681d

Means 5368a 4787b 4550c

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 45 decrease in the serum LDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

120

50864735

4216

537651435345

49424567

53655388

41614495

30

35

40

45

50

55

60

Base Line Week-2 Week-4

Weeks

LDL

(mg

dl)

A B C D

Figure 46 Effect of β-glucan beverage on LDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

121

beverage containing 06 β -glucan (D) exhibited the lowest

content of LDL (4681 mgdl) in serum of adult subjects I t is

evident from Figure 46 that concentration of LDL decreased

progressively by increasing the level of β -glucan in the beverages

The level of LDL content decl ined at a faster rate in case of

beverages C (04 β -glucan) and D (06 β -glucan) as compared

to control beverages (0 β -glucan) The LDL concentration

decreased at higher rate during f irst two weeks as compared to

the last two weeks of the experimental study I t is also evident

from Figure 45 that at the end of two weeks of study period the

highest decrease in LDL (1082) content was observed in the

subjects group when the data for beverages pooled

The decrease in LDL content was recorded at faster rate during

1s t two weeks of study The beverage showed maximum response

towards decrease LDL content in the beginning of the study as

compared to the last weeks of the study period (Figure 46)

Braaten et a l (1994) have reported 10 decrease in LDL

cholesterol concentrations in hypercholesterolemic men and

women who consumed daily for 4 weeks 72 g of oat gum

containing 58 g of β -glucan mixed with a noncarbonated drink or

with water Kahlon and Chow (1997) also found similar results in

hyperl ipidaemic subjects fed on oat water-soluble gum These

f indings are well in support of the present results in which a

decrease in LDL level by the intake of β -glucan in the functional

beverage formulations

122

464 High Density Lipoproteins (HDL)

The analysis of variance regarding serum HDL level of adult

subjects showed signif icant effect of beverages and study periods

on HDL content (Table 426) The interaction between beverages

and study periods was observed to be non signif icant for this HDL

content of serum

The results i l lustrated in Figure 48 and Table 430 showed a

variable response by different functional beverages towards level

of HDL in different groups of people The serum HDL content was

recorded higher in the subjects fed on D beverage (06 β -glucan)

while the lowest HDL content was recorded in the group fed on

control beverage (0 β -glucan) (Fig48) I t is also evident from

Figure 47 that higher increase in level of tr iglyceride was

observed by the increasing level of β -glucan in the formulation of

different beverages

The HDL content increased at a faster rate during f irst two

weeks while the rate of increase was less at the end of the

experimental study The highest increase in the HDL content was

observed in the group fed on the beverage D (06 β -glucan) when

tested at the end of week 2 while the lowest increase was

observed in the group consuming control beverage The increase

in HDL content of test subjects was lower after fol lowing f irst two

weeks of study

123

Week2Week4

135

532

9931069

005025034 0310

123456789

1011

In

crea

se

Study Period

ABCD

Table 430 Effect of β-glucan supplemented beverage on serum HDL content (mgdl) of healthy subjects

Study Periods Beverage

Base Line Week-2 Week-4 Means

A 6237 6321 6324 6261d

B 6184 6513 6529 6398c

C 6206 6822 6845 6608b

D 6214 6878 6899 6632a

Means 6210c 6634a 6580b

Means carrying same letters within a column or row do not differ significantly (P lt 001)

Figure 47 increase in the serum HDL level of subjects fed on different beverages

A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

124

62246184

6497

6237 6321

65136206

67956822 6803

6214

6878

58

60

62

64

66

68

70

Base Line Week-2 Week-4

Weeks

HDL

(mg

dl)

A B C D

Figure 48 Effect of β-glucan beverage on HDL (mgdl) content of healthy

volunteers A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

125

The study period showed a signif icant effect on the HDL

content of test subjects The maximum increase in HDL was

observed in the f irst f i f teen days (two week) while the lower

response was observed following the next f i f teen days upto the

expiry of the experiment (Table 430) The results of the present

study are well supported by Kalra and Jood (2000) who observed a

higher HDL content of rats with the consumption of barley β -

glucan gum as compared to control group of rats The results from

such type of studies demonstrated that every 1 rise in HDL by

the uti l ization of medicine there is a 3 reduction chance in

coronary heart diseases (Frick et a l 1987) The results of the

present study are also in l ine with the f indings of Naumann et a l

(2006) who incorporated β -glucan into fruit drink and observed

274 percent increase in HDL during f ive weeks study period in

human subjects They suggested that in order to overcome and

reduce cardiovascular diseases i t is better to use β-glucan in our

daily diet because low HDL heightened risk for heart disease The

results of the present study showed that intake of β -glucan in

beverage signif icantly reduced serum cholesterol and LDL while

signif icantly increased HDL level This study demonstrates that

beverage containing β-glucan can help to reduce risk of coronary

heart disease

465 Blood Glucose concentarion

The statist ical results regarding blood glucose level of adult

volunteers showed signif icant effect of β -glucan treatment

feeding intervals and study periods on blood glucose level (Table

432) The interactive effect of intervals and treatments also

126

possessed signif icant effect on the blood glucose of adult

volunteers subjects All interactions among these three variables

were found to be non signif icant for blood glucose level

The results presented in Table 433 showed different

response towards level of blood glucose by different beverages I t

is evident from the results (Table 432) that higher blood glucose

level (10017 mgdl) was observed in the adults fed on control

beverage i e A (0 β -glucan) fol lowed by beverage B (02 β -

glucan) The lowest blood glucose content (9755 mgdl) was

recorded in the group fed with D beverage (06 β -glucan) i t is

also obvious from the results shown in Figure 49 that higher

reduction in blood glucose level of adult subjects was observed by

increasing the level of β -glucan in the beverage formulation The

level of blood glucose increased in al l beverages t i l l f irst hour of

study and then started declining after one hour The results

indicated (Table 433) that rate of reduction in the concentration

of blood glucose was signif icantly different among different

beverages The adult subjects fed on beverages D (06 β -glucan

beverage) showed higher reduction in blood glucose level than

groups fed on al l other treatments The blood glucose level of the

adults fed with beverage D reduced from 9339 mgdl to 8135

mgdl from 0 to 60 minutes of the study

The blood glucose level varied signif icantly during different

study periods I t is evident from Table 432 that blood glucose

was found the highest (9510 mgdl) at the beginning of the study

(0 day) when the data for beverage and study period were pooled

but i t reduced signif icantly from 9324 mgdl to 9192 mgdl

127

Table 431 Mean sum of squares for blood glucose contents of volunteers SOV df MSS Intervals (A) 5 12929373 Diets (B) 3 19069863 Days (C) 2 17178671 A x B 15 94341233 A x C 10 26435555NS B x C 6 15218384 NS A x B x C 30 13125518 NS Error 144 18758931 Total 215

Table 432 Effect of β-glucan beverage on blood glucose (mgdl)content

with different time intervals Beverage Days 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

day0 8533 10132 11045 10875 10533 10141 day15 8401 9813 10833 10629 10348 9841

A day30 8246 9927 10637 10426 10217 9725

day0 8499 9862 10662 10330 10034 9430 day15 8360 9860 10432 10020 9730 9355 B

day30 8219 9823 10414 9766 9650 9212 day0 8518 9220 9643 9445 9149 8445

day15 8363 9273 9520 9336 8880 8319 C day30 8250 9026 9461 9242 8727 8267

day0 8520 9202 9502 9288 8977 8261 day15 8374 9051 9319 8846 8732 8152 D day30 8215 8921 9212 8684 8350 7993

Table 433 Interactive effect of diets and time scale intervals on the blood glucose

contents (mgdl) of volunteers Time scale intervals Beverage 0 Min 30 Min 60 Min 90 Min 120 Min 180 Min Means

A 8393 9957 10838 10643 10366 9903 10017a B 8359 9848 10503 10039 9805 9333 9648b C 8377 9173 9541 9341 8919 8344 8949c D 8370 9058 9344 8939 8686 8135 8755d

Means 8375e 9509c 10057a 9741b 9444c 8929d 0 Min = fasting

128

Effect of different beverages on the blood glucose level of subjects

60

70

80

90

100

110

120

0 Min 30 Min 60 Min 90 Min 120 Min 180 Min

Time (Minutes)

mg

dl

Diet A

Diet B

Diet C

Diet D

Figure 49 Effect of β-glucan beverage on blood glucose (mgdl) content of

healthy volunteers Table 434 Interactive effect of diets and study duration on the blood glucose

contents (mgdl) of volunteers Beverage Study Periods

0 Days 15 Days 30 Days Means

A 10210 9978 9863 10017a B 9803 9626 9514 9648b C 9070 8949 8829 8949c D 8958 8746 8562 8755d

Means 9510a 9324b 9192c A = Beverage supplemented with 0 β-glucan (control) B = Beverage supplemented with 02 β-glucan C = Beverage supplemented with 04 β-glucan D = Beverage supplemented with 06 β-glucan

129

when blood glucose level was observed after 15 and 30 days

respectively

The interactive effect of diets (beverages) and study t ime

(Table 432) indicated that the control beverage (0 β -glucan)

possessed the highest blood glucose level of adults when tested

f irst t ime however the lowest blood glucose level was observed

in the adult subjects who were fed on diet D (06 β -glucan

beverage) when tested after 30 days (Table 432)

The results indicated that level of blood glucose was

signif icantly affected by the difference in beverages and t ime

intervals The beverages supplemented with β -glucan showed

pronounced effect on the reduction of blood glucose level

whereas the control diet did not signif icantly affect the level of

blood glucose in the adult subjects The reduction in blood

glucose level was more when level of β -glucan in the beverage

formulations was increased I t is true due to the assumption that

complex carbohydrates were digested and absorbed more slowly

than simple sugars result ing in a f lattened glucose response

curve The fal lacy was revealed when researchers discovered that

blood glucose and insulin responses varied greatly independent

of diet c lassif ication as simple or complex carbohydrate

(Schauberger et a l 1977 Jenkins et a l 1983)

The β -glucan has abil i ty to retard the absorption rate of food

in the intest ine due to increased viscosity thus balancing the

post-prandial glucose and insulin response (Wursch and Sunyer

1997 Wood et a l 2000) The viscous nature of β -glucan physically

slows glucose absorption in the gut This property is useful in the

130

formulation of products targeting management of diabetes Wood

et a l (1990 and 1994) also reported similar results who prepared

porridge from βndashglucan and after consumption demonstrated that

product has reduced postprandial blood glucose level Jenkins et

a l (2002) showed that a food in which β -glucan is incorporated as

a functional ingredient tends to reduce glycemic indices of that

particular food addition of β -glucan predictably reduces the GI

while maintaining palatabil i ty Foster-Pwer and Miller (1994) also

observed similar reduction in blood glucose level by the β -glucan

containing food bars Thus the reduction of blood glucose in the

present study by intake of beverages containing β -glucan is in l ine

with the f indings reported above I t may be concluded from the

present study that diabetic patient may use beverages in which β -

glucan is incorporated which wil l help to reduce the level of

blood glucose

131

CHAPTER-5

SUMMARY

Barley (Hordeum vulgare L) is one of the f irst ancient plant

species I t is r ich in dietary f ibre and possessing mixed-l inkage

(1rarr3) (1rarr4)-β -D-glucans a soluble f iber component The

nutrit ional and functional properties of β -glucan make it suitable

ingriedient to use in functional foods The β -glucan was used for

the development of functional beverages and the results are

summarised as follow

The barley f lour contained crude protein crude fat crude

f iber ash and nitrogen free extract (NFE) 1165 231 675

222 and 7707 respectively The barley f lour possessed total

dietary f ibre (TDF) and β -glucan content 1148 and 487

respectively The crude protein crude fat crude f iber ash and

nitrogen free extract (NFE) in β -glucan was found 9 96 117

722 172 and 7638 respectively The β -glucan contained

soluble dietary f iber (SDF) insoluble dietary f iber (IDF) and a

total dietary f iber (TDF) 7505 1025 and 8530 respectively

The β -glucan possessed 263 pentosans The crude fat and ash

contents in β -glucan gum pellets were found 117 and 172

respectively

The L-value (color index) of functional beverages increased

signif icantly as the level of β -glucan increased in the formulation

of different beverages The beverage of T6 containing 10 β -

132

glucan showed the highest L-value (2128) and fol lowed by

control beverage (without β -glucan) which got L-value 1969 L-

value of functional beverages declined signif icantly as the storage

period increased

The beverage of T5 containing 08 β -glucan gave the

highest a-value (165) and the lowest a-value (-227) was given

by T1 control beverage (without β -glucan) a-value of functional

beverages decreased signif icantly by increasing in storage

intervals b-value was signif icantly affected by treatments as well

as storage intervals The beverage T1 contains 02 pectin

possessed the highest b-value (1080) fol lowed by the beverage

T6 contains 1 β -glucan and signif icantly the lowest b-value was

recorded in the beverage of T2 (02 β -glucan)

The viscosity of beverages improved signif icantly due to the

incorporation of β -glucan in beverages The highest viscosity

(2175 mPa-s) was found in beverages of T6 containing 1 β -

glucan fol lowed by T5 beverage containing 08 β-glucan The

lowest viscosity was recorded in beverage of T1 (0 β -glucan)

The total soluble solids were signif icantly affected by the levels of

β -glucan in beverages The highest of total soluble solids

(1042ordmbrix) were yielded by the the beverages of T6 containing 1

β -glucan fol lowed by beverage of T5 containing 08 β -glucan T1

(0 β-glucan) gave the lowest total soluble solids (TSS) The pH

of different beverages differed signif icantly due to storage

intervals The pH decreased signif icantly in al l beverages

throughout the storage period Total acidity and ascorbic acid

varied signif icantly as a function of storage The ascorbic acid

content was higher (29406 mgkg) in fresh beverage which

133

declined signif icantly to 27933 mgkg and 26211 mgkg after 45

and 90 days of storage respectively Reducing sugars showed non

signif icant change due to incorporation of β -glucan in different

beverage The reducing sugars increased from 372 to 431 from 0

to 90 days of storage respectively The non reducing sugars

differed signif icantly among different beveragesThe total plate

count (TPC) values decreased in al l beverages during the storage

periods The TPC value of freshly prepared beverages (0 day) was

higher 129 times 104 - 4 46 times 104 which decreased to 117 times 104 - 4 32 times

104 at the end of the storage

The color scores differed signif icantly due to treatments and

storage intervals among beverages The beverage containing 02

β -glucan got the highest color scores (684) fol lowed by the

control (0 2 pectin) while beverage of (1 0 β -glucan) got the

lowest scores (494) The scores of f lavor varied signif icantly due

to differences (β -glucan levels) in treatments as well as storage

intervals The beverage of T3 containing 04 β -glucan got

signif icantly the highest scores for f lavor The highest scores for

sweetness (674) were given to control beverage fol lowed by

beverage containing 02 β -glucan The lowest scores (503) was

given to the sourness of T6 beverage (10 β -glucan) The scores

given to sourness of beverages decreased as a function of storage

period

The beverage prepared from the control treatment T2 (02

Pectin) got the highest total scores (731) The beverage containing

more than 06 of β -glucan got mimimum total scores for overall

acceptabil i ty Total scores among beverages decreased

signif icantly among storage periods

134

Total serum cholesterol of the test subjects was affected

signif icantly due to variation in beverage formulations and study

periods Maximum total cholesterol (13953 mgdl) was recorded

in the control group and the lowest content of total cholesterol

(13230 mgdl) in serum of adult subjects was observed when

human subjects were fed on 06 β -glucan The contents of total

serum cholesterol decreased signif icantly by increasing the level

of β -glucan in the beverages Minimum decrease decrease in the

serum cholesterol was measured in the test group fed on control

beverage (0 β -glucan)

The level of serum triglyceride was found higher in the human

subject fed on control beverage (0 β -glucan) and the lowest

tr iglyceride content was observed in the subjects fed on beverage

D (06 β -glucan) Higher reduction in the tr iglyceride content

was found by increasing the level of β -glucan in the beverage

formulations Maximum decrease in tr iglyceride content (1099)

was recorded in the subject group fed on the beverage D (06 β -

glucan)

The highest concentration of LDL (5202 mgdl) was found

in the human subject group fed on control beverage The beverage

containing 06 β -glucan (D) exhibited the lowest content of LDL

(4681 mgdl) in serum of the test subjects The LDL decreased

progressively by increasing the level of β -glucan in the beverage

formulations The serum HDL content was observed higher in the

human subjects fed on D beverage (06 β -glucan) while the

lowest HDL content was recorded in the human fed on control

beverage (0 β -glucan)

135

The blood glucose level of human subjects was affected

signif icantly by treatments feeding intervals and study periods

Higher blood glucose level (10017 mgdl) was observed in the

adults fed on control beverage i e A (0 β -glucan) and fed on

beverage B (02 β -glucan) The lowest blood glucose content

(9755 mgdl) was measured in the human subject group fed on D

beverage (06 β -glucan) Higher reduction in blood glucose level

was observed by increasing the level of β -glucan in the beverage

formulations The rate of reduction in the concentrat ion of blood

glucose was signif icantly different for different functional

beverages The human subjects fed on beverage D (06 β -glucan

beverage) showed higher reduction in level of blood glucose than

groups fed on al l other beverages The blood glucose level of the

adults fed on beverage D reduced from 9339 mgdl to 8135

mgdl during 0 to 60 minutes of the study

I t is evident from the present study that (1rarr3) (1rarr4) - β -D-

glucan is a dominant soluble f iber component in barley During

three months refrigerated storage barley β -glucan was found to be

stable at low pH conditions in beverages system and showed shelf

stabil i ty Consumption of foods rich in β -glucan (soluble f iber)

may reduce the risk of chronic diseases and such foods exhibited

decrease in serum cholesterol levels and postprandial blood

glucose levels in adult subjects This study suggested the use of β -

glucan in beverages can help to reduce riskes of coronary heart

disease and diabetes

136

Conclusions

Concentration of β -glucan had a signif icant effect on the

sensory parameters of beverage

Beverage formulate with the incorporation of β -glucan exert

i ts effect on physicochemical characterist ics of beverage

β -glucan improved most of the sensory characterist ics of the

beverage

The beverages below 08 containing β -glucan were found to

be acceptable during the three month refrigerated storage

period

The different formulated functional beverages showed no

phase separation very minute quantity of impurit ies such as

protein and starch content founded at the bottom of bott les

All levels of β -glucan decrease the total cholesterol LDL

cholesterol and triglycerides in healthy subjects

Further research is needed to know the thermal stabil i ty of

β -glucan and its behavior with other food ingredients in

beverages application to make stable foods

137

Recommendations

All local and indigenous sources for β -glucan isolation should be exploited

The relationship between molecular weight of β -glucan with respect to physiological functional i ty has to be kept in mind

Clinical studies are needed to investigate the physiological effects of β -glucan preparations differing in molecular weight and viscosity

Studies should be carried out to explore the molecular weight of β -glucan to proper understanding of functional properties of β -glucan

Consumer studies are needed to explore the acceptabil i ty of food products having β -glucan along with the substitution of β -glucan enriched barley f lour for some wheat f lour and dairy products

There is need to develop new foods with the addition of soluble dietary f iber from barley source with enhanced health properties by keeping in mind shelf stabil i ty

Structural differences which are present in the soluble and insoluble dietary f ibre of β -glucan should also be investigated for indigenous variet ies

The Genes responsible for the synthesis of β -glucan should be characterized and identif ied in cereal crops and strains of microorganisms

The role of β -glucan in increasing immune system should also be discovered

138

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Aastrup S 1979a The effect of rain on β -glucan content in barley grains Carlsberg esearch Communications 44381-393

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Anderson J W and J Tieyen-clark 1986 Dietary f iber Hyperlipidemiahypertension and coronary heart disease Am J Gastroenterol 81907-919

Anderson J W DB Spencer CC Hamilton SF Smith and J Tietyen CA Bryant P Oeltgen 1990 Oat-bran cereal lowers serum total and LDL cholesterol in hypercholesterolemic men Am J Clin Nutri 52 495-499

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Andersson AAM E Armo E Grangeon H Fredrikssonm RA Andersson P Man 2004 Molecular weight and structure units of (1- 3 1-4)- β -glucans in dough and bread made from hull- less barley mil l ing fractions J Cereal Sci 40195ndash204

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Assmann G 1979 HDL-cholesterol precipitant Randox Labs Ltd CrumLin Co Antrim N Ireland Internist 20559-567

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Basman A and HK Ksel 1999 Properties and composit ion of Turkish f lat bread (bazlama) supplemented with barley f lour and wheat bran Cereal Chem 76506ndash511

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Beer MU PJ Wood J Weisz N Fi l l ion 1997 Effect of cooking and storage on the amount and molecular weight of (1rarr3) (1rarr4) - β -D-glucan extracted from oat products by an in vitro digestion system Cereal Chem 74 705-709

Bell S VM Goldman BR Bistrian AH Arnold G Ostroff R Forse 1999 Effect of β -glucan from oats and yeast on serum lipids Crit Rev Food Sci Nutri 39(2) 189ndash202

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Beneke ES 1962 Medical Mycology Lab Manual Burgess Pub Co Minneapolis Minnisota USA

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Bhatty R S 1999 The potential of hull- less barley Cereal Chem 76(5) 589ndash599

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Biorklund M Rees A van RP Mensink and G Onning 2005 Changes in serum lipids and postprandial glucose and insulin concentrations after consumption of beverages with β -glucan from oat or barley a randomized dose-controlled tr ial Eur J Clin Nutri 591272-1281

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BNF (Brit ish Nutrit ion Foundation) 1994 Starchy Foods in the Diet BNF London

Braaten J T PJ Wood FW Scott MS Wolynetz MK Lowe P Bradleywhite MW Coll ins 1994 Oat β -glucan reduces blood cholesterol concentration in hypercholesterolemic subjects Eur J Clin Nutri 48465ndash474

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Brennan C S and LJ Cleary 2005 The potential use of cereal (13 14)-b-D-glucans as functional food ingredients J CerSci 421ndash13

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Brennan CS CM Tudorica V Kuri 2002 Soluble and insoluble dietary f ibres (non-starch polysaccharides) and their effects on food structure and nutrit ion F Ind J 5 261-272

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Brunswick P DJ Manner and J K Stark 1987 Development of β -D-glucanases during germination of barley and the effect of ki lning on individual isoenzymes J Inst Brew 93181-186

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Burkus Z 1996 Barley P-Glucan Extraction Functional Properties and Interactions with Food Components MSc thesis Edmonton AlbertaCanda

Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloid F Glicksman M (Ed) p 49-93 CRC Press Inc Boca Raton

Burkus Z 1996 Barley β -glucan Extraction Functional properties and interaction with food components MSc Thesis Dept of Agricultural Food and Nutrit ional Science Univ of Alberta Edmonton Canada

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Burkus Z and F Temeil i 1998 Effect of extraction conditions on yield composit ion and viscosity stabil i ty of barley P-glucan gum Cer Chem 75 805-809

Burkus Z and F Temell i 1999 Glucan concentrate J Food Sci 64198-201 Glicksman M 1982 Functional properties of hydrocolloids Ch 3 in Food Hydrocolloidr Glicksman M (Ed) p 49-93 CRC Press hc Boca Raton FL

Burkus Z and F Temell i 2005 Rheological properties of barley β -glucan Carbohydr Polym 59 459ndash465

Burkus Z F Temell i 1999 Gelation of barley β -glucan - concentrate J Food Sci 64198-201

Calix FD and N Bardrie 2004 Consumer acceptance and physicochemical quality of processed red sorrelroselle (Hibiscus sabdar i f fa L) sauces from enzymatic extracted calyces 4 141-148

Carpita NC 1996 Structure and biogenesis of cel l walls of grasses Annual Rev Plant Physiol Plat Molecular Biol 47445-476

Carr J M S Glatter J L Jeraci and B A Lewis 1990 Enzymes Determination of Beta-Glucan in Cereal-Based Food Products Cereal Chem 67226-229

Casterl ine J L CJ Oles and Y Ku 1997 In vitro fermentation of various food f iber reactions J Agric Food Chem 452463ndash2467

Cavallero S F Empill i Brighenti and A M Stanca 2002 High (1rarr31rarr4)-_-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response J Cere Sci 36 59ndash66

Chowdhury MGF MN Islam MS Is lam T Is lam and MS Hossain 2008 Study on Preparation and Shelf-Life of Mixed Juice Based on Wood Apple and Papaya J Soil Nature 2(3) 50-60

Chung OK and Y Pomeranz 1985 Amino acids in cereal proteins and protein fractions Ch 5 in Digesfibi l i~ and

144

Amino Acid Availabil i ty in Cereals andOilseeds J W Finley and DT Hopkins (Eds) pp 169-232 AACC St Paul MN

Clara C J Mar ıacutea Esteve and Ana Fr ıacutegola 2008 Color of orange juice treated by High Intensity Pulsed Electric Fields during refrigerated storage and comparison with pasteurized juice Food Control 19 151ndash158

Crandall PG CS Chen and KC Davis 1987 Preparation and storage of 72 brix orange juice concentration J Food Sci 52 (3) 381

Davidson MH andm A McDonald 1998 Fiber forms and functions Nutri Res 18 617ndash624

Daw ZY YSA El-Gizaw and AMB Said 1994 Microbiological evaluation of some local juices and drinks Chemie Mikrobiologie Technologie der Lebensmittel 168ndash15

Dawkins N L and I D Nnanna 1995 Composit ion molecular 4)-3 1A 1995 Studies on oat gum [(1 weight est imation and rheological properties Food Hydrocol 9 1-7

Dawkins NL I A Nnanna 1993 Studies on oat gum [(1rarr31rarr4)- β-D-glucan] Composit ion molecular weight est imation and rheological properties Food Hydrocol 9 1-7

Del PS F Leonett i DC Simonson P Sheehan M Matsuda and RA DeFronzo 1994 Effect of sustained physiologic hyperinsulinaemia and hyperglycaemia on insulin secretion and insulin sensit ivity in man Diabetologia 371025ndash1035

Delaney B RJ Nicolosi TA Wilson T Carlson S Frazer GH Zheng R Hess K Ostergren J Haworth and N Knutson 2003 The American Society for Nutrit ional Sciences J Nutri 133468-475

DeVries J W 2001 AACC report The definit ion of dietary f iber Cereal Foods World 46(3) 112-126

Dohnalek MH 2004 The role of f ibre in cl inical nutrit ion In Van der Kamp JW Asp NG Miller J J Schaafsma G (Ed) Dietary f ibre bioactive carbohydrates for food and feed Wageningen Academic Publishers Wageningen pp 271294

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Dongowski G M Huth E Gebhardt and W Flamme 2002 Dietary f iber-rich barley products beneficial ly affect the intestinal tract of rats J Nutri 132(12) 3704-14

Drzikova B G Dongowski E Gebhardt and A Habel 2005 The composit ion of dietary f ibre-rich extradites from oat affects bi le acid binding and fermentation in vitro Food Chem 90 181-192

Estevea MJ A Fr ıgola C Rodrigob and D Rodrigo 2005 Effect of storage period under variable conditions on the chemical and physical composit ion and colour of Spanish refrigerated orange juices Food and Chemical Toxicol 431413ndash1422

Etoh H K Murakami T Yogoh H Ishikawa Y Fukuyama and H Tanaka 2004 Antioxidative compounds in barley tea Biosci Biotechnol Biochem 682616-2618

Falade OS OR Sowunmi A Oladipo A Tobosun and SRA Adewusi 2003 The level of organic acids in some Nigerian fruit and their effect on mineral availabil i ty in composite diet Pak J Nutri 2(2) 82-83

Faraj A T Vasanthan R Hoover 2006 The influence of a-amylase-hydrolysed barley starch fractions on the viscosity of low and high purity barley b-glucan concentrates Food Chem 9656ndash65

Fasoyiro S B OA Ashaye A Adeola and FO Samuel 2005 Chemical and Storabil i ty of Fruit-Flavoured (Hibiscus sabdariffa) Drinks World J Agric Sci 1(2) 165-168

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Foster-Powell K J B Mil ler 1994 International tables of glycaemic index Am J Clin Nutr 59 66ndash 69

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Frick MH O Elo and K Haapa 1987 Helsiniki heart study Primary prevention tr ial with germfibrozil in middle aged men with dyslipidemia N Eng J Med 3171237-45

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Fuleki T E Pelayo and RB Palabay 1994 Sugar composit ion of varietal juices produce from fresh and stored apple J Agric Food Chem 42 1266-75

Gallaher DD CA Hassel 1995 The role of viscosity in the cholesterol lowering effect of dietary f iber In Kritchevsky D Bonfield C editors Dietary f iber in health and disease Minnesota Eagan Press 106-114

Gasiorowski H H Chalcarz A Aniola J I Nahrung 2000 Mil l ing of barley to obtain beta-glucan enriched products Aug 44(4) 238-41

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Granzer R 1982 changes in fruit juices in consumer packs during extended storage Verpackungs-Rundschau 33(6) 35-4

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Hall ikainen MA ES Sarkkinen MI J Uusitupa 2000 Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose-dependent manner J Nutri 30 767ndash776

Hancioglu O and M Karapinar 1997 Microflora of boza a tradit ional fermented Turkish beverage Int J Food Microbiol 35271ndash274

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Handan E S Celik B Bi lgi and H Koksel 2005 A new approach for the uti l ization of barley in food products Food Chemistry1-7 Received 6 December 2004received in revised form 7 March 2005accepted 7 March 2005

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Helm CV and A Francisco 2004 Chemical characterization of Brazil ian hulless barley variet ies f lour fractionation and protein concentration Scientia Agricola 61593-97

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Holtekjolen AK AK Uhlen E Brathen E Brathen S Sahlstrom and SH Khnutesen 2006 Contents of starch and non-starch polysaccharides in barley variet ies of different origin Food Chem 94348 -358

Izydorczyk M S J Symons and J E Dexter 2002 Fractionation of wheat and barley In L Marquart J L Slavin amp R G Fulcher (Eds) Whole grain foods in health and disease (pp

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Izydorczyk MS A Hussain AW MacGregor 2001 Effect of barley and barley components on rheological properties of wheat dough J Cer Sci 34251ndash260

Izydorczyk MS LJ Macri AW MacGregor 1998a Structure and physicochemical properties of barley non-starch polysaccharides-I Water-extractable beta-glucans and arabinoxylans Carbo Poly 35249ndash258

Izydorczyk MS LJ Macri AW MacGregor 1998b Structure and physicochemical properties of barley non-starch polysaccharides-II Alkali-extractable beta-glucans and arabinoxylans Carbo Poly 35 259ndash269

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Jaumlrvi AE BE Karlstroumlm YE Granfeldt I ME Bjoumlrck NG Asp and BOH Vessby 1999 Improved glycemic control and l ipid profi le and normalized f ibrinolytic activity on a lowglycemic index diet in type 2 diabetic patients Diabetes Care 2210ndash18

Jaskari J K Henriksson A Nieminen T Suortt i H Salovaara K Poutanen 1995 Effect of hydrothermal and enzymic treatments on the viscous behaviour of dry- and wet-milled oat barns Cereal Chem 72625-631

Jenkins AL DJ Jenkins U Zdravkovic P Wursch and V Vuksan 2002 Depression of the glycemic index by high