incorporation of rice starch affecting on morphology, mechanical properties and water vapor...

INCORPORATION OF RICE STARCH AFFECTING ONMORPHOLOGY, MECHANICAL PROPERTIES AND WATER VAPORPERMEABILITY OF GLUTELIN-BASED COMPOSITE FILMSDOUNGJAI THIRATHUMTHAVORN1 and WIRAWAN THONGUNRUAN

Department of Food Technology, Faculty of Engineering and Industrial Technology, Silpakorn University, 6 Rachamakanai Road, Mueng district,Nakhon Pathom 73000, Thailand

1Corresponding author.TEL: 663-421-9361;FAX: 663-427-2194;EMAIL: [email protected]

Received for Publication August 1, 2012Accepted for Publication June 13, 2013

doi:10.1111/jfpp.12149

ABSTRACT

Protein is a by-product of rice starch production. The major component of riceprotein found in endosperm is glutelin, an alkali-soluble protein. Pure glutelinfilm could not peel off from the casting plate; however, glutelin films containingstarch were easily removed from the plate. Various ratios of rice starch and glutelinat 100:0, 92:8, 75:25, 50:50 and 25:75 (w/w) were applied for casting film. Sorbitolwas used as a plasticizer. Morphology was studied by scanning electron micro-scope. The mechanical properties and water vapor permeability (WVP) wereevaluated. The results were found that rice starch and glutelin-based compositefilms became thinner with smoother surface as starch content increased. Elonga-tion and flexibility of composite films containing glutelin could be improved byincorporation of rice starch at a considerable amount without affecting on tensilestrength and WVP.

PRACTICAL APPLICATIONS

This research had an objective to study the surface morphology, mechanical prop-erties and WVP of composite films based on glutelin (by-products from produc-ing rice starch) and rice starch. Those properties are important for theapplications used as edible films/coatings. Edible films and coatings could be usedto protect the product from mechanical damage, physical and chemical. In thispaper, we did not add any antimicrobial agent. So, this film cannot protect theproduct from microbiological activities. The applications of edible films andcoating have been successfully applied in fresh foods and processed foods.

INTRODUCTION

Edible films and coatings could be used to protect theproduct from mechanical damage, physical, chemical andmicrobiological activities. The structural materials areclassified as proteins, lipids, polysaccharides or composite(Falguera et al. 2011). Proteins and polysaccharides providegood mechanical and organoleptic properties and are effec-tive barriers to aroma compounds, light and gases such asoxygen and carbon dioxide (Xu et al. 2005; Bourtoom andChinnan 2008). Starch films and coatings are odorless,tasteless, colorless, nontoxic, biologically absorbable (Cutterand Sumner 2002), semipermeable to carbon dioxide(Guilbert et al. 1996; Garcia et al. 2001; Cutter and Sumner2002) and good oxygen barriers (Mark et al. 1966; Roth andMehtretter 1967), with many characteristics similar to

plastic films (Lourdin et al. 1995; Rindlav-Westling et al.1998). However, starch films and coatings are tacky athigh relative humidity and rather brittle at extremely lowhumidities (Jokay et al. 1967).

Proteins have been widely studied as film-forming mate-rials due to their relative abundance, film-forming abilityand nutritional qualities (Park et al. 2002).

Protein films appear to have lower tensile strength (TS)than most polysaccharide films. Proteins can be combinedwith polysaccharides to modify film mechanical properties.A polysaccharide material, such as starch, can be combinedwith protein to produce a blended film system. It appearsthat there is little effect on film water vapor permeability(WVP), but that addition of polysaccharide may increasefilm oxygen permeability and TS and decrease film elonga-tion (E) (Krochta 2002).

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Journal of Food Processing and Preservation 38 (2014) 1799–1806 © 2013 Wiley Periodicals, Inc. 1799

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There are many reports on composite films made bymixing protein with polysacchride for example; rice proteinconcentrate/pullulan (Shih 1996); whey protein isolate(WPI)/mesquite gum (Oses et al. 2009); WPI/pullulan(Gounga et al. 2007); casein/starch (Jagannath et al. 2003);soy protein/starch (Zeng et al. 2011); soy protein isolate(SPI)/carboxylmethyl cellulose (Su et al. 2010); SPI/propyleneglycol alginate (PGA) (Shih 1994); SPI/PGA(Rhim et al. 1999); SPI/cellulose (Wu et al. 2009); soybeanflour protein/pectin (Liu et al. 2007); gelatin/starch(Arvanitoyannis et al. 1998; Jagannath et al. 2003); gelatin/sago starch (Al-Hassan and Norziah 2012); gelatin/gellan(Lee et al. 2004); gelatin/pectin (Liu et al. 2007); gelatin/chitosan (Cheng et al. 2003); egg white/dialdehyde starch(Gennadios et al. 1998); albumen/starch (Jagannath et al.2003; Wongsasulak et al. 2006); zein/starch (Zeng et al.2011); gliadin/starch (Zeng et al. 2011). The interaction ofproteins and polysaccharides produced better strength andmoisture barrier film properties (Shih 1994). The associa-tion among the polymers can be achieved through blend-ing, laminating or coating with other polymers withdesirable properties. Blending is an easier and more effectiveway to prepare compatible multiphase polymeric materials(Zhong and Xia 2008).

Rice is the most widely consumed basic food in theworld. The Office of Agricultural Economics (2012)reported that Thailand could produce about 22 milliontons per year. Rice grain consists of rice starch (90%) andothers such as protein (7–8%), fat (0.4–0.6%), fiber (0.3–0.6%) and ash (0.4–0.9%). Rice starch is a promisingmaterial for making film because of its low cost andrenewability, as well as possessing good mechanical proper-ties (Xu et al. 2005). The blending of starch and proteincan improved water barrier of the films and increased TS(Jagannath et al. 2003).

Rice proteins are considered valuable because they arecolorless, rich in essential amino acids, possess a bland taste,and are hypoallergenic and hypocholesterolemic. Glutelin, amajor protein found in rice endosperm, can be extractedfrom regular rice flour by alkali method (Shih and Daigle2000). Glutelin of rice is a protein of high molecular weight(6 × 104–6 × 105) and composed of subunits bound bydisulphide linkages (Tecson et al. 1971; Ju et al. 2001). Thedenaturation temperature of glutelin was 82.2C (Ju et al.2001). The most abundant amino acids in rice glutelin areglutamine, asparagine, arginine, glycine and alanine. Theamide groups in glutamine and asparagine side chainspromote aggregation of glutelin (Wen and Luthe 1985;Paraman et al. 2007). Glutelin is extremely insoluble inwater because of hydrophobic, hydrogen and disulfidebonding (Agboola et al. 2005). There are some researchesregarding rice protein films that are the effect of pH ondefatted rice bran films (Gnanasambandam et al. 1997) and

the composite films based on rice protein concentrate andpullulan (Shih 1996).

This research had an objective to study the surface mor-phology, mechanical properties and WVP of compositefilms based on rice starch and glutelin.

MATERIALS AND METHODS

Materials

Glutelin was extracted from rice flour by modified methodof Ju et al. (2001). Rice flour was first defatted by hexane.Rice proteins, albumin, globulin and prolamin were sequen-tially extracted as described (Ju et al. 2001). After that, glute-lin was extracted using 0.1 N NaOH and then precipitatedby adjusting pH to 4.8 (an isoelectric point of glutelin). Theextracted glutelin contained 64.39% protein and 4.72% car-bohydrate. Native rice starch (1.08% protein) and rice flour(7.53% protein) were from Cho Heng Co., Ltd., NakhonPathom, Thailand. Sorbitol was purchased from Siam Sor-bitol Co., Ltd., Bangkok, Thailand. Sodium hydroxide was ofAmerican Chemical Society reagent grade.

Methods

Film Preparation. Aqueous dispersions of rice starch orglutelin were prepared. Rice starch (3% w/w dry basis oftotal solution) and sorbitol (40% w/w dry basis of totalsolid) were dispersed in distilled water for 5 min. Glutelin(3% w/w dry basis of total solution) was dissolved in0.1 M NaOH for 1 h. Each dispersions were prepared in thefollowing proportions of rice starch to glutelin (100:0, 92:8,75:25, 50:50 and 25:75 w/w). The mixture was stirred at500 rpm for 5 min using magnetic stirrer. The pH was thenadjusted to 11 with 0.1 M NaOH. After that, the mixturewas heated to 80C for 20 min on a hot plate and thendegassed. The solution was poured onto plastic plate anddried at 40 ± 1C for 16 h in a convection oven (model WTB,Binder, Tuttlingen, Germany). The films were peeled manu-ally and stored in a closed container at 25 ± 5C and 50 ± 5%relative humidity (RH) for 2 days for further evaluation.

Film Properties. Surface Morphology. The dried filmsamples were mounted on a metal stub with double-sidedadhesive tape and sputter coated with a layer of gold priorto imaging. The morphological structures of the films werestudied by a JSA-541QLV (SEM, JEOL Ltd., Tokyo, Japan)and the images were taken at accelerating voltage 10 kV anda magnification 1,000 times of origin specimen size.

Color and Opacity. The samples were cut 20 mm wideand 20 mm long. The color and opacity of film sampleswas determined with a colorimeter (HunterLab, model

COMPOSITE FILMS BASED ON GLUTELIN AND RICE STARCH D. THIRATHUMTHAVORN and W. THONGUNRUAN

Journal of Food Processing and Preservation 38 (2014) 1799–1806 © 2013 Wiley Periodicals, Inc.1800

Miniscan XE, Reston, VA), working with D65 (day light) andmeasure cell with opening of 30 mm, using the CIELabcolor parameters. The color of films was expressed (Eq. (1))as the difference of color (ΔE*).

Δ Δ Δ ΔE L a b* * * *= ( ) + ( ) + ( )2 2 2 (1)

Where ΔL*, Δa*, Δb* are the differentials between the colorparameter of the samples and the color parameter of thewhite standard (L* = 93.64, a* = −1.19, b* = 1.09) used asthe film background. The opacity (Y) was calculated as theratio between the opacity of each sample on the black stan-dard (Yb) and the opacity of each sample on the white stan-dard (Yw). Three replicates of each film sample weredetermined randomly for Yb and Yw. The results wereexpressed as: Y = Yb/Yw.

Film Thickness. Film thickness was measured at five differ-ent locations with a micrometer (No. 7326, MitutoyoManufacturing, Kanagawa, Japan) to the nearest 0.0001inch (0.0025 mm.).

Mechanical Properties. TS, %E at break and elastic modulus(EM) were determined by Texture analyzer (TA-XT2i, StableMicro System Co, Ltd., Surrey, U.K.). Film specimens werecut as a rectangular center, 15 mm wide and 60 mm longand then preconditioned at 25 ± 5C and 50 ± 5% RH for atleast 24 h. Initial grip separation and the crosshead speedwere set at 30 mm and 60 mm/min, respectively. TS was cal-culated by dividing the maximum load for breaking the filmby its cross-sectional area. %E was determined by dividingthe film E at rupture by the initial grip separation. EM wasdefined as the ratio of stress to strain in the initial linearpart of the stress–strain curve.

WVP. WVP was determined by modified method ofNavarro-Tarazaga et al. (2008). Three specimens from eachreplication of each formulation were cut and mounted onpolymethacrylate test cups containing 6 mL of distilledwater. Thickness of each film was measured with a microm-eter at five randomly selected points before the film wasattached to the cup. The specimens were placed in a desicca-tor cabinet at 25 ± 2C and 0% RH using anhydrous calciumsulfate. Weights were taken periodically until steady wasachieved, and average film thickness measured at fiverandom positions was used to calculate the resulting WVP.

RESULTS AND DISCUSSION

Surface Morphology

Starch films containing glutelin appeared rougher surfacethan pure starch film (Fig. 1). The incorporation of increas-

ing amounts of glutelin in the formulation resulted in filmswith greater roughness. This was related to the unfoldedproteins (glutelin) that underwent the aggregation throughhydrogen, ionic, hydrophobic and covalent bonding(Prodpran and Benjakul 2005). The protein-protein interac-tion in the film samples increases with a large proportion ofprotein; causing the higher aggregation and yields filmswith rougher surface (Oses et al. 2009). Excessive hydropho-bic and disulphide interactions of rice protein were theprime cause of protein insolubility (Paraman et al. 2007). Inaddition, the presence of polysaccharide in the film contain-ing protein enhanced protein aggregation (Oses et al. 2009).

The compatibility of polymers can be evaluated throughthe microstructural analysis (Liu et al. 2007; Jiménez et al.2012; Zhang et al. 2013). A continuous phase of theglutelin-starch films without noticeably different phases wasobserved, indicating that both polymers are compatible.The similar results have been reported in other blendedpolymers, such as gelatin-starch (Zhang et al. 2013), pectin-fish skin gelatin (Liu et al. 2007) and pectin-soybean flourprotein (Liu et al. 2007).

Color and Opacity

The b* values markedly increased with the addition of moreglutelin, that is films became more yellow. Therefore, theyaffected to the larger ΔE* values. The yellowish color of thefilms would be related to the presences of proteins in theircomposition (Tapia-Blacido et al. 2007). However, L* valueswere not significantly different (P > 0.05) (Table 1). Filmscontaining glutelin presented more yellow because of thecomplex formation between protein and polyphenolic com-pound in alkali conditions (Gnanasambandam et al. 1997).This is mainly due to alkalinity and heat reaction during thefilm preparation (Jangchud and Chinnan 1999; Bamdadet al. 2006). Alkaline solvents can extract pigments morethan other solvents (Bamdad et al. 2006).

Rice starch-based film had less opacity than rice starchmixed with glutelin-based films (Table 1). The addition ofglutelin at higher amount in rice starch-based edible filmsleaded to the greater interaction between protein molecules.The aggregated proteins randomly dispersed in the starchmatrix, and as such, exist in film as a discontinuous phaseafter drying. Therefore, proteins provide more interfacialarea within a film structure. The light diffusion is increasedand results in a reduction in film transparency (Thys et al.2008).

Thickness

The incorporation of glutelin at high proportion in the for-mulation resulted in the thicker films (Table 2). This was

D. THIRATHUMTHAVORN and W. THONGUNRUAN COMPOSITE FILMS BASED ON GLUTELIN AND RICE STARCH


a b

dc

eFIG. 1. SCANNING ELECTRON MICROGRAPHS OF COMPOSITE FILMS BASED ON RICE STARCH AND GLUTELIN AT 100:0 (A), 92:8 (B), 75:25 (C),50:50 (D) AND 25:75 (E) (W/W)



related to the denatured proteins that can be increasinglyreconnect to network or aggregation more than protein-starch interaction and distributed throughout the films asseen in scanning electron micrographs (Fig. 1).

Mechanical Properties

Films must be generally resistant to breakage and abrasion(to strengthen the structure of a food filling, to protect itand to ease handling) and flexible (enough plasticity toadapt to possible deformation of the fillings without break-ing) (Gontard and Guilbert 1994). TS is the maximum TSwhich a material can sustain. E is as a measure of the film’sability to stretch and an indication of the films’ flexibilityand stretchability (extensibility). EM is a measure of theintrinsic stiffness of the film (Robertson 1993; Bourtoomand Chinnan 2008). Plasticizers are used to overcome starchfilm brittleness and improve flexibility and extensibility(Arvanitoyannis et al. 1997; Sothornvit and Krochta 2005).Sorbitol is one of the plasticizers commonly used toimprove the flexibility of starch films. It has the advantageof improving mechanical properties, with less increase inpermeability compared to other plasticizers (Mchugh andKrochta 1994; Mchugh et al. 1994; Sothornvit and Krochta2000, 2001; Laohakunjit and Noomhorm 2004). Therefore,sorbitol was used as a plasticizer in this study. In addition,the type of film-forming material affects the mechanicalproperties of edible or biodegradable films depending onthe structure of the polymer and especially molecular

length, geometry, molecular weight distribution and thetype and position of its lateral groups (Guilbert andGontard 1995).

Pure rice starch-based films exhibited the greatest TS andE at break (Table 2). The fact that film with just starch wasfounded to have the greatest TS, suggests that intermolecu-lar forces between the starch chains were the responsible ofthe film strength.

The films containing higher amount of glutelin exhibitedmore rigid film. With the addition of more glutelin, filmstrength decreased and so did the brittleness. The additionof glutelin into the film formulation reduced flexibility andincreased stiffness. It indicated that rice starch-based filmscontaining glutelin were stiffer that related to molecularentanglement and interactions (Zhong and Xia 2008; Suet al. 2010). The denaturation of the protein promotes theinteraction between protein chains by intermoleculardisulphide and hydrogen bonds, resulting in a rigid matrixwith low E (Mchugh et al. 1994; Mariani et al. 2009; Oseset al. 2009). Protein has a tendency to keep its rigid andstrong structure, being made of disulfide and hydrogenbond (Mariani et al. 2009). The same trend was found inthe films made from carboxymethyl cellulose/SPI (Su et al.2010) and mesquite gum/WPI (Oses et al. 2009).

WVP

WVP remained unchanged for films with glutelin concen-tration as high as 50% (Fig. 2). This was due to the larger

TABLE 1. THE L*, A*, B*, ΔE* AND OPACITY OF COMPOSITE FILMS BASED ON RICE STARCH AND GLUTELIN AT VARIOUS RATIOS

Starch : Glutelin L* a* b* ΔE* Opacity

100:0 90.15 ± 0.94a1/ (–)1.30 ± 0.05d 1.54 ± 0.09e 3.50 ± 1.05c 2.16 ± 0.76c

92:8 90.68 ± 0.96a (–)1.52 ± 0.07c 2.37 ± 0.19d 3.81 ± 0.56c 3.36 ± 0.86bc

75:25 89.05 ± 1.05a (–)1.66 ± 0.01bc 3.67 ± 0.29c 5.33 ± 0.74b 3.63 ± 0.64b

50:50 89.55 ± 0.77a (–)1.78 ± 0.08ab 5.47 ± 0.85b 6.21 ± 0.93ab 4.57 ± 0.82ab

25:75 89.07 ± 0.42a (–)1.81 ± 0.03a 6.65 ± 0.59a 7.23 ± 0.72a 4.92 ± 0.65a

1/ Means values with different letters in the same column are significantly different (P < 0.05).

TABLE 2. THICKNESS, TENSILE STRENGTH (TS), % ELONGATION AT BREAK (%E) AND ELASTIC MODULUS (EM) OF COMPOSITE FILMS BASED ONRICE STARCH AND GLUTELIN AT VARIOUS RATIOS

Starch :Glutelin Thickness (μm) TS (MPa) %E EM (MPa)

100:0 65.29 ± 4.32b1/ 4.14 ± 0.37a 149.74 ± 14.36a 26.22 ± 3.66bc

92:8 67.83 ± 5.02b 3.38 ± 0.30b 144.52 ± 10.78ab 18.04 ± 3.35c

75:25 68.58 ± 4.08ab 3.77 ± 0.40ab 124.24 ± 15.26b 29.65 ± 6.63bc

50:50 70.28 ± 3.61ab 3.02 ± 0.54b 92.93 ± 8.77c 46.35 ± 8.74b

25:75 74.00 ± 7.40a 3.46 ± 0.51b 50.51 ± 10.67d 83.04 ± 17.12a

1/ Means values with different letters in the same column are significantly different (P < 0.05).



number of hydrophilic interactions in the starch/glutelinfilms enhances water diffusion through the film (Su et al.2010). Polysaccharide and/or protein components of ediblefilms are characterized by an emphatic hydrophilicity, whichis responsible for film-water interactions and structuralchanges in the polymeric film network (Kristo et al. 2007).

The resistant to water vapor decreased, with increasingglutelin concentration. This may relate to the aggregation ofprotein that causes less compact structure. The addition ofglutelin in the rice starch/glutelin composite films at highconcentration may decrease the intermolecular interactionsand increase the electrostatic repulsion between starch andglutelin, thus increasing WVP. The results were similar tosome composite films i.e., rice protein isolate/pullulan (Shih1996); SPI/konjac glucomanan/chitosan films (Bourtoomand Chinnan 2008).

CONCLUSION

The incorporation of glutelin on starch films had a signifi-cant effect on the mechanical properties with less effect onits WVP. For films with about a ratio of glutelin and ricestarch at 25:75, improvements could be achieved withacceptable film strength and practically no sacrifice in watervapor resistance.

ACKNOWLEDGMENTS

This research work was supported by Research and Devel-opment Institute, Silpakorn University, Thailand.

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incorporation of rice starch affecting on morphology, mechanical properties and water vapor...

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