honey based polyherbal anti tussive and expectorant …

www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │

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Asadulla et al. World Journal of Pharmacy and Pharmaceutical Sciences

HONEY BASED POLYHERBAL ANTI TUSSIVE AND EXPECTORANT

AND THEIR ANTI-BACTERIAL ANALYSIS

Syed Asadulla*, Champa Kumari S., Fathima Fayiza, Fathimath Fayiza M., Fathima

Unaisa U.K., Fayis Muhammed K. and Mintu Aleyas

Department of Pharmacognosy, Malik Deenar College of Pharmacy, Kasaragod, Kerala.

ABSTRACT

The extracts of dried powders of Lobelia (Lobelia erinus),

Pushkaramoola root (Inula racemosa), Tulsi (Ocimum sanctum),

vasaka(Adhatoda vasika), Yastimadhu (Glycyrrhiza glabra), Long

pepper(Pipper longum) etc and also plants which enhance thier anti

microbial activity like cardamom, mustard, cinnamon, kantakari, etc

was selected in the present study for developing poly herbal syrup. The

prepared polyherbal syrup was evaluated immediately after

preparation. The purpose of this research was to formulate honey

polyherbal antitussive and expectorant and to evaluate antimicrobial

activity of these preparation againt S.aureus and E.coli by using agar

well diffusion method using different concentration of extracts.

KEYWORDS: Polyherbal, formulation, syrup, liquid dosage form, S.aureus, E.coli.

INTRODUCTION

There are many types of drugs that are used to suppress cough and are often prescribed in

combination. Before dealing with the particular type of drug used, it is important to consider

briefly the nature of cough production, its role in disease and desirability of suppressing it.

The studies showing that about 95 million units of pediatric drugs were sold for cough and

cold every year in India. Commercially available cough syrups containing synthetic and

semisynthetic have several disadvantages. To overcome such disadvantages naturally

occurring herbs having antitussive and expectorant activity can be used individually or in

combination. The herbal medicines are normally considered safer than the non -herbal

medicines because the naturally active ingredients in the herbal medicines are in combination

with other component.

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 7.632

Volume 9, Issue 11, 1768-1779 Research Article ISSN 2278 – 4357

*Corresponding Author

Syed Asadulla

Department of

Pharmacognosy, Malik

Deenar College of

Pharmacy, Kasaragod,

Kerala.

Article Received on

06 Sept. 2020,

Revised on 27 Sept. 2020,

Accepted on 18 October 2020

DOI: 10.20959/wjpps202011-17686


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Cough syrups are the medicated liquid preparations used for relieve coughing. Cough syrups

can be prepared by using traditional drugs. The herbal cough syrups are available in the

market and Are more effective against cough. In the recent times the use of herbal cough

syrups is on the rise due to the spread of the awareness of the effect of complementary and

alternative medicine. It is also due to the much stronger believe that alternative therapy is

with less side effects, that makes the demand of herbal products in the market. People are

willing to follow alternative therapy since nowadays they are well bothered about the side

effects of organic medicines.

Herbal cough syrups are prepared by using honey as vehicle. The stability and fragrance of

the preparation can be improved by addition of excepients. Herbal formulations were

generally examined through physicochemical as well as pharmacological studies. pH,

viscosity, skin irritations, diffusion studies, visual appearance and stability studies.

Combination of plant drug extract shows synergistic activities like antimicrobial activity.

METHODOLOGY

Plant Collection

The plants were collected from Bangalore. The plant material was taxonomically identified

by the botanist Mr. Dr. V S Anil Kumar, H O D, department of botany, Govt college

Kasaragod. The plant parts were dried and then powdered with mechanical grinder and stored

in an air tight container.

PREPARATION OF EXTRACT

Maceration

Weighed about 25 g of powdered crude drug, to this added 100 ml of water and 10 g of

sucrose(10% of water taken) and boiled for 2 hrs. Allowed to cool to this added 1.5 g of yeast

and kept for 7 days. Then the mixture were filtered through muslin cloth and evaporated to

get the extract

Preparation of Artificial Honey

For the preparation of honey first we have taken the microorganism Aspergillus niger. Which

is obtained from the bulbs of Onion as well as garlic. It can be seen in the decade bulbs in the

form of black powder. It is then collected and cultured using Sabouraud’s dextrose agar

medium. The fully grown fungi were obtained after 5 days.


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In this honey we are giving a pineapple flavor so that the palatability can be enhanced. For

that we have taken a large pineapple, it is then cut in to small pieces, blended it and allowed

to boil for the separation of fibers about 2-3 hrs. Then the juice was filtered using muslin

cloth.

For the artificial honey we have to prepare immobilized Aspergillus beads. This beads can be

prepared by following procedure, 2% of sodium alginate solution was prepared by dispersing

and mixing 1 g in 50 ml of Tris buffer HCl. The Aspergillus separated from culture medium

was added to 50 ml of alginate solution and mixed thoroughly to make dispersion. The gel

was then taken into a syringe and added drop wise to calcium chloride solution of volume 50

ml. It was then kept for 20 minutes and beads obtained were washed with dist. Water and

then with buffer. It was then dried to room temperature.

Here we tried to prepare the best honey for that we have taken different concentration of

pineapple juice as well as sucrose using 6 beakers. Each beaker was added with 100 beads of

Aspergillus. To this 90 ml, 85 ml, 80 ml, 75 ml, 70 ml, 66.4 ml pine apple juice was added

from 1-6 beakers. Similarly sucrose was added in concentration of 10%, 15%, 20%, 25%,

30%,33.6% respectively. Then the beakers were kept for 7 days and observed regularly.

The best honey was obtained in the 6 th beaker which contained high amount of fructose. It

was then taken and filtered through muslin cloth for the separation of beads. Then it was

warmed and allowed to cool. Now the artificial honey is ready to use.

1) 90 ml juice + 10% sucrose 2) 85 ml juice + 15% sucrose


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3)80 ml juice + 20% sucrose 4) 75 ml juice +25% sucrose

(5) 70 ml juice + 30% sucrose (6) 66.6 ml juice+ 33.4% sucrose

PREPARATION OF POLYHERBAL ANTITUSSIVE AND EXPECTORANT

Formulation For Antitussive

Aniseed 10mg

Thyme 10mg

Peppermint 30mg

Cinnamon 15mg

Pushkaramoola root .................................. 250mg

Kantakari plant 150mg

Ocimum sanctum ...................................... 250mg

Vasaka leaf 250mg

Yeshtimadhu stem ..................................... 250mg

Shunti root 250mg

Pippal root 250mg

Pineapple flavoured honey .......................................... qs


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Formulation For Expectorant

Lobelia 50mg

Liquorice 100mg

Wild cherry 15mg

Black mustard seed…… 15mg

Black pepper 30mg

Cardamom 30mg

Ammonium chloride ............................... 0.138g

sod. citrate 57.03mg

Menthol 1.14mg

Nilgiri oil 1.2mg

Pineapple flavoured honey ............................. qs

Procedure

Sufficient quantity of drug extract were weighed and dissolved in small amount of prepared

honey. These are mixed well and add preservative and other additives in remaining amount of

honey. And mix the two solutions. Volume was made to 10 ml with honey.

Physico Chemical Evaluation of Anti Tussive And Expectorant

The physical parameters of antitussives and expectorants like colour, appearance, pH were

evaluated.

Visual Appearance

The preparations were inspected for its clarity, color and transparency. They were also

evaluated for the presence of particles.

Stability Study

The purpose of stability testing is to provide evidence on how the quality of drug substance

varies with time, under the influence of variety of factors such as temperature, humidity and

light. And to establish a retest period for the drug substance or a shelf life for the drug

product and recommended storage conditions.

Placebo and cough syrup were evaluated for the thermo stability, 10 ml of the antitussive an

10 ml of expectorant was taken in 25 ml beaker, each were taken in 2 beakers, one was kept

at room temperature and the other was kept at 40˚C for 2 weeks. It was also examined for pH,


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appearance and homogenecity.

p H STUDIES

Well calibrated pH meter was used to determine the pH of cough syrups. The result were

documented and compared over 2 weeks.

Antibacterial Activity Test

Individual plant extracts and formulated antitussive and expectorant are studied for its anti-

microbial properties. However there are no reports regarding the evaluation of antimicrobial

activity of these herbs in combination. Hence the drug was evaluated for antimicrobial

activity in S. aureus and E. coli

Screening of Antibacterial Activity

Table No 2: Screening Of Antibacterial Activity Preparation Of Culture Media

Preparation Of Nutrient Agar Media.

ORGANISM S. aureus, E. coli

METHOD Agar well diffusion method

MEDIA Nutrient agar media

STANDARD Ciprofloxacin

TEST Plant extracts and formulations

Suspended 1.4 g in 50 ml dist. Water, heat to boiling to dissolve the medium completely.

Sterilized by autoclaving at 15 Ibs pressure. 121ºC for 15 minutes

Screening By Agar Well Diffusion Method

Nutrient agar was prepared. Then it was poured on sterilized petri plate and allowed to

solidify. The bacterial culture was swabbed over the plates. Then, the wells were prepared by

using sterilized cork borer. To that the separate drug extract and combined drug extract as

well as std ciprofloxacin was added. Then the plates were incubated at 37˚C for 24-48 hrs.

The zone of inhibition was measured in mm.

Confirmation of Fructose Content In Honey

For the confirmation of fructose content in honey seliwanoff’s test and Osazone test was done

Seliwanoff’s Test

To 3 ml of seliwanoff’s reagent, add 1 ml of the test solution. Boil in water bath for 2

minutes. A cherry red colour precipitate with in 5 minutes was obtained.


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Osazone Test

To 5 ml of test solution add 10 drops of glacial acetic acid 1 g of crystalline sodium acetate,

0.5 g of phenyl hydrazine hydrochloride and mix well. Keep the tube boiling for 20 minutes

and cool and observed under microscope.

RESULT AND DISCUSSION

Plant Collection and Drying

Dried plant parts used for the study was collected from Bangalore, Karnataka and some from

Kasaragod too. The drugs were identified and authenticated by Dr. V.S Anil kumar, HOD,

Govt. college, Kasaragod.

Preparation of Extract

The extract were prepared by maceration. The extract were evaporated. The percentage yield

of extracts were calculated and result are shown below

Drugs % Yield of Extracts

Lobelia 13.8%

Kantakari 9%

Aniseed 9.7%

Black mustard 8.5%

Black pepper 7%

Vasaka 10%

Tulsi 8.7%

Pepper mint 10.2%

Liquorice 12.5%

Pushkaramoola 11%

Ginger 6.8%

Thyme 6.6%

Wild cherry 10.8%

Cinnamon 11.4%

Cardamom 10%

Long pepper 9%

Formulation of Polyherbal Anti Tussive And Expectorant

The poly herbal antitussive and expectorant were prepared on the basis of the formula given

Visual Appearance

When the optimized formula was evaluated for 4 weeks the visual appearance of the

formulations were found to be unchanged, stable and acceptable.


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Stability Study

The formulations were stored under and observed under room temperature and elevated

temperature (40˚C) for 4 weeks. On observation the formulations were found to be stable,

unchanged and good in appearance. Hence it was found to be thermo stable.

Table: stability study.

Formulation Appearance Homogenicity Ph

Anti tussive Stable and

unchanged Maintained 4

Expectorant Stable and unchanged Maintained 4.2

Evaluation of Antibacterial Activity

Table: evaluation of antibacterial activity.

Sample Concentration(µl)

Zone of inhibition(mm)

Bacterial culture

S. aureus E. coli

Lobelia

50 10 mm 11 mm

75 12 mm 13mm

100 13 mm 15mm

Liquorice

50 9 mm 12 mm

75 10mm 12mm

100 11 mm 13 mm

Kantakari

50 10 mm 10 mm

75 11 mm 12 mm

100 12 mm 13 mm

Ginger

50 10 mm 11 mm

75 12 mm 13mm

100 13 mm 14 mm

Cardamom

50 8 mm 10 mm

75 11mm 10 mm

100 10 mm 13 mm

Pushkaramoola

50 11mm 14 mm

75 12mm 15mm

100 14mm 15 mm

Vasaka

50 9mm 10mm

75 11 mm 12mm

100 11mm 13mm

Pepper mint

50 +9mm 10mm

75 11mm 12mm

100 13mm 14mm

Black pepper

50 8mm 9mm

75 10mm 11mm

100 11mm 12mm

Black mustard 50 10mm 12mm

75 11mm 13mm


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100 11mm 14mm

Long pepper

50 9mm 10mm

75 10mm 12mm

100 9mm 10mm

Aniseed

50 10mm 13mm

75 11mm 14mm

100 14mm 15mm

Thyme

50 8mm 9mm

75 10mm 10mm

100 11mm 13mm

Tulasi

50 10mm 12mm

75 13mm 14mm

100 14mm 15mm

Wild cherry

50 7mm 8mm

75 9mm 9mm

100 10mm 11mm

Cinnamon

50 10mm 12mm

75 11mm 13mm

100 12mm 14mm

Anti tussive 100 15 mm 19 mm

Expectorant 100 18 mm 21 mm

Standard Ciprofloxacin 20mm 25 mm

Pushkaramoola Aniseed Mustard Thulasi Ginger

Cardomom Kantakari Thyme Liquorice Black pepper


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Lobelia Pepper mint Vasaka Wild cherry Long pepper

Cinnamon

Anti Bacterial Effect of Formulation

A- S. aureus B-E,coli C- S. aureus D-E. coli

A – Antibacterial activity of antitussive against S. aureus

Antibacterial activity of antitussive against E. coli

C - Antibacterial activity of Expectorant against S. aureus

D - Antibacterial activity of Expectorant against E. coli

FORMULATIONS

Pineapple honey Expectorant Antitussive


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Identification of Fructose Content In Honey

By molisch’s and fehling’s test it was found that the prepared pineapple flavored honey

contains fructose. For the confirmation also done the osazone test and the seliwanoff’s test.

The result of osazone test is given below

Cherry red Broom stick

CONCLUSION

The present study aimed at the formulation of Honey based polyherbal anti tussive and

expectorant and their Anti- bacterial analysis

Extraction of various crude drugs were done by Soxhlet extraction technique (methanolic

extract), and maceration technique (aq.extract).

Polyherbal antitussive and expectorant were prepared using honey as the solvent, menthol,

thymol, Ammonium chloride, it yielded a formulation of satisfactory phytochemical

appearance.

The stability study of the formulation was carried out.

Studied the Anti-bacterial activity of the individual extract as well as the formulation against

S. aureus and E. coli.

As per our research work, it reveals that the drug extracts have a good Antibacterial activity,

How ever further studies to be done to improve the stability of the formulations.

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