Journal of Food Processing and Preservation ISSN 1745-4549

INCORPORATION OF RICE STARCH AFFECTING ON

MORPHOLOGY, MECHANICAL PROPERTIES AND WATER VAPOR
PERMEABILITY OF GLUTELIN-BASED COMPOSITE FILMS
DOUNGJAI 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

1
Corresponding author. ABSTRACT
TEL: 663-421-9361;
FAX: 663-427-2194; Protein is a by-product of rice starch production. The major component of rice
EMAIL: thdoungjai@yahoo.com protein found in endosperm is glutelin, an alkali-soluble protein. Pure glutelin
film could not peel off from the casting plate; however, glutelin films containing
Received for Publication August 1, 2012
starch were easily removed from the plate. Various ratios of rice starch and glutelin
Accepted for Publication June 13, 2013
at 100:0, 92:8, 75:25, 50:50 and 25:75 (w/w) were applied for casting film. Sorbitol
doi:10.1111/jfpp.12149 was used as a plasticizer. Morphology was studied by scanning electron micro-
scope. The mechanical properties and water vapor permeability (WVP) were
evaluated. The results were found that rice starch and glutelin-based composite
films became thinner with smoother surface as starch content increased. Elonga-
tion and flexibility of composite films containing glutelin could be improved by
incorporation of rice starch at a considerable amount without affecting on tensile
strength 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 the
applications used as edible films/coatings. Edible films and coatings could be used
to protect the product from mechanical damage, physical and chemical. In this
paper, we did not add any antimicrobial agent. So, this film cannot protect the
product from microbiological activities. The applications of edible films and
coating have been successfully applied in fresh foods and processed foods.

plastic films (Lourdin et al. 1995; Rindlav-Westling et al.

INTRODUCTION
Edible films and coatings could be used to protect the
product from mechanical damage, physical, chemical and
microbiological activities. The structural materials are
classified as proteins, lipids, polysaccharides or composite
(Falguera et al. 2011). Proteins and polysaccharides provide
good mechanical and organoleptic properties and are effec-
tive barriers to aroma compounds, light and gases such as
oxygen and carbon dioxide (Xu et al. 2005; Bourtoom and
Chinnan 2008). Starch films and coatings are odorless,
tasteless, colorless, nontoxic, biologically absorbable (Cutter
and Sumner 2002), semipermeable to carbon dioxide
(Guilbert et al. 1996; Garcia et al. 2001; Cutter and Sumner
2002) and good oxygen barriers (Mark et al. 1966; Roth and
Mehtretter 1967), with many characteristics similar to
1998). However, starch films and coatings are tacky at
high relative humidity and rather brittle at extremely low
humidities (Jokay et al. 1967).
Proteins have been widely studied as film-forming mate-
rials due to their relative abundance, film-forming ability
and nutritional qualities (Park et al. 2002).
Protein films appear to have lower tensile strength (TS)
than most polysaccharide films. Proteins can be combined
with polysaccharides to modify film mechanical properties.
A polysaccharide material, such as starch, can be combined
with protein to produce a blended film system. It appears
that there is little effect on film water vapor permeability
(WVP), but that addition of polysaccharide may increase
film oxygen permeability and TS and decrease film elonga-
tion (E) (Krochta 2002).

Journal of Food Processing and Preservation 38 (2014) 1799–1806 © 2013 Wiley Periodicals, Inc. 1799
COMPOSITE FILMS BASED ON GLUTELIN AND RICE STARCH
There are many reports on composite films made by
mixing protein with polysacchride for example; rice protein
concentrate/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); soybean
flour 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 of
proteins and polysaccharides produced better strength and
moisture barrier film properties (Shih 1994). The associa-
tion among the polymers can be achieved through blend-
ing, laminating or coating with other polymers with
desirable properties. Blending is an easier and more effective
way to prepare compatible multiphase polymeric materials
(Zhong and Xia 2008).
Rice is the most widely consumed basic food in the
world. The Office of Agricultural Economics (2012)
reported that Thailand could produce about 22 million
tons per year. Rice grain consists of rice starch (90%) and
others 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 promising
material for making film because of its low cost and
renewability, as well as possessing good mechanical proper-
ties (Xu et al. 2005). The blending of starch and protein
can improved water barrier of the films and increased TS
(Jagannath et al. 2003).
Rice proteins are considered valuable because they are
colorless, rich in essential amino acids, possess a bland taste,
and are hypoallergenic and hypocholesterolemic. Glutelin, a
major protein found in rice endosperm, can be extracted
from regular rice flour by alkali method (Shih and Daigle
2000). Glutelin of rice is a protein of high molecular weight
(6 × 104–6 × 105) and composed of subunits bound by
disulphide linkages (Tecson et al. 1971; Ju et al. 2001). The
denaturation temperature of glutelin was 82.2C (Ju et al.
2001). The most abundant amino acids in rice glutelin are
glutamine, asparagine, arginine, glycine and alanine. The
amide groups in glutamine and asparagine side chains
promote aggregation of glutelin (Wen and Luthe 1985;
Paraman et al. 2007). Glutelin is extremely insoluble in
water because of hydrophobic, hydrogen and disulfide
bonding (Agboola et al. 2005). There are some researches
regarding rice protein films that are the effect of pH on
defatted rice bran films (Gnanasambandam et al. 1997) and
1800 Journal of Food Processing and Preservation 38 (2014) 1799–1806 © 2013 Wiley Periodicals, Inc.
D. THIRATHUMTHAVORN and W. THONGUNRUAN
the composite films based on rice protein concentrate and
pullulan (Shih 1996).
This research had an objective to study the surface mor-
phology, mechanical properties and WVP of composite
films based on rice starch and glutelin.
MATERIALS AND METHODS
Materials
Glutelin was extracted from rice flour by modified method
of 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 precipitated
by adjusting pH to 4.8 (an isoelectric point of glutelin). The
extracted 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., Nakhon
Pathom, Thailand. Sorbitol was purchased from Siam Sor-
bitol Co., Ltd., Bangkok, Thailand. Sodium hydroxide was of
American Chemical Society reagent grade.
Methods
Film Preparation. Aqueous dispersions of rice starch or
glutelin were prepared. Rice starch (3% w/w dry basis of
total solution) and sorbitol (40% w/w dry basis of total
solid) were dispersed in distilled water for 5 min. Glutelin
(3% w/w dry basis of total solution) was dissolved in
0.1 M NaOH for 1 h. Each dispersions were prepared in the
following proportions of rice starch to glutelin (100:0, 92:8,
75:25, 50:50 and 25:75 w/w). The mixture was stirred at
500 rpm for 5 min using magnetic stirrer. The pH was then
adjusted to 11 with 0.1 M NaOH. After that, the mixture
was heated to 80C for 20 min on a hot plate and then
degassed. The solution was poured onto plastic plate and
dried 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 film
samples were mounted on a metal stub with double-sided
adhesive tape and sputter coated with a layer of gold prior
to imaging. The morphological structures of the films were
studied by a JSA-541QLV (SEM, JEOL Ltd., Tokyo, Japan)
and the images were taken at accelerating voltage 10 kV and
a magnification 1,000 times of origin specimen size.
Color and Opacity. The samples were cut 20 mm wide
and 20 mm long. The color and opacity of film samples
was determined with a colorimeter (HunterLab, model
D. THIRATHUMTHAVORN and W. THONGUNRUAN
Miniscan XE, Reston, VA), working with D65 (day light) and
measure cell with opening of 30 mm, using the CIELab
color parameters. The color of films was expressed (Eq. (1))
as the difference of color (ΔE*).
ΔE * = ( ΔL*)2 + ( Δa*)2 + ( Δb*)2
Where ΔL*, Δa*, Δb* are the differentials between the color
parameter of the samples and the color parameter of the
white standard (L* = 93.64, a* = −1.19, b* = 1.09) used as
the film background. The opacity (Y) was calculated as the
ratio 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 were
determined randomly for Yb and Yw. The results were
expressed as: Y = Yb/Yw.
Film Thickness. Film thickness was measured at five differ-
ent locations with a micrometer (No. 7326, Mitutoyo
Manufacturing, Kanagawa, Japan) to the nearest 0.0001
inch (0.0025 mm.).
Mechanical Properties. TS, %E at break and elastic modulus
(EM) were determined by Texture analyzer (TA-XT2i, Stable
Micro System Co, Ltd., Surrey, U.K.). Film specimens were
cut as a rectangular center, 15 mm wide and 60 mm long
and then preconditioned at 25 ± 5C and 50 ± 5% RH for at
least 24 h. Initial grip separation and the crosshead speed
were set at 30 mm and 60 mm/min, respectively. TS was cal-
culated by dividing the maximum load for breaking the film
by its cross-sectional area. %E was determined by dividing
the film E at rupture by the initial grip separation. EM was
defined as the ratio of stress to strain in the initial linear
part of the stress–strain curve.
WVP. WVP was determined by modified method of
Navarro-Tarazaga et al. (2008). Three specimens from each
replication of each formulation were cut and mounted on
polymethacrylate test cups containing 6 mL of distilled
water. Thickness of each film was measured with a microm-
eter at five randomly selected points before the film was
attached to the cup. The specimens were placed in a desicca-
tor cabinet at 25 ± 2C and 0% RH using anhydrous calcium
sulfate. Weights were taken periodically until steady was
achieved, and average film thickness measured at five
random positions was used to calculate the resulting WVP.
RESULTS AND DISCUSSION
Surface Morphology
Starch films containing glutelin appeared rougher surface
than pure starch film (Fig. 1). The incorporation of increas-
Journal of Food Processing and Preservation 38 (2014) 1799–1806 © 2013 Wiley Periodicals, Inc.
COMPOSITE FILMS BASED ON GLUTELIN AND RICE STARCH
ing amounts of glutelin in the formulation resulted in films
with greater roughness. This was related to the unfolded
proteins (glutelin) that underwent the aggregation through
hydrogen, ionic, hydrophobic and covalent bonding
(Prodpran and Benjakul 2005). The protein-protein interac-
(1) tion in the film samples increases with a large proportion of
protein; causing the higher aggregation and yields films
with rougher surface (Oses et al. 2009). Excessive hydropho-
bic and disulphide interactions of rice protein were the
prime cause of protein insolubility (Paraman et al. 2007). In
addition, the presence of polysaccharide in the film contain-
ing protein enhanced protein aggregation (Oses et al. 2009).
The compatibility of polymers can be evaluated through
the microstructural analysis (Liu et al. 2007; Jiménez et al.
2012; Zhang et al. 2013). A continuous phase of the
glutelin-starch films without noticeably different phases was
observed, indicating that both polymers are compatible.
The similar results have been reported in other blended
polymers, such as gelatin-starch (Zhang et al. 2013), pectin-
fish skin gelatin (Liu et al. 2007) and pectin-soybean flour
protein (Liu et al. 2007).
Color and Opacity
The b* values markedly increased with the addition of more
glutelin, that is films became more yellow. Therefore, they
affected to the larger ΔE* values. The yellowish color of the
films would be related to the presences of proteins in their
composition (Tapia-Blacido et al. 2007). However, L* values
were not significantly different (P > 0.05) (Table 1). Films
containing glutelin presented more yellow because of the
complex formation between protein and polyphenolic com-
pound in alkali conditions (Gnanasambandam et al. 1997).
This is mainly due to alkalinity and heat reaction during the
film preparation (Jangchud and Chinnan 1999; Bamdad
et al. 2006). Alkaline solvents can extract pigments more
than other solvents (Bamdad et al. 2006).
Rice starch-based film had less opacity than rice starch
mixed with glutelin-based films (Table 1). The addition of
glutelin at higher amount in rice starch-based edible films
leaded to the greater interaction between protein molecules.
The aggregated proteins randomly dispersed in the starch
matrix, and as such, exist in film as a discontinuous phase
after drying. Therefore, proteins provide more interfacial
area within a film structure. The light diffusion is increased
and 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
1801
COMPOSITE FILMS BASED ON GLUTELIN AND RICE STARCH D. THIRATHUMTHAVORN and W. THONGUNRUAN

a b

c d

e
FIG. 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)

1802 Journal of Food Processing and Preservation 38 (2014) 1799–1806 © 2013 Wiley Periodicals, Inc.
D. THIRATHUMTHAVORN and W. THONGUNRUAN COMPOSITE FILMS BASED ON GLUTELIN AND RICE STARCH

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 ± 0.94 a1/
(–)1.30 ± 0.05 d
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).

related to the denatured proteins that can be increasingly
reconnect to network or aggregation more than protein-
starch interaction and distributed throughout the films as
seen 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 it
and to ease handling) and flexible (enough plasticity to
adapt to possible deformation of the fillings without break-
ing) (Gontard and Guilbert 1994). TS is the maximum TS
which a material can sustain. E is as a measure of the film’s
ability to stretch and an indication of the films’ flexibility
and stretchability (extensibility). EM is a measure of the
intrinsic stiffness of the film (Robertson 1993; Bourtoom
and Chinnan 2008). Plasticizers are used to overcome starch
film brittleness and improve flexibility and extensibility
(Arvanitoyannis et al. 1997; Sothornvit and Krochta 2005).
Sorbitol is one of the plasticizers commonly used to
improve the flexibility of starch films. It has the advantage
of improving mechanical properties, with less increase in
permeability compared to other plasticizers (Mchugh and
Krochta 1994; Mchugh et al. 1994; Sothornvit and Krochta
2000, 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 mechanical
properties of edible or biodegradable films depending on
the structure of the polymer and especially molecular
length, geometry, molecular weight distribution and the
type and position of its lateral groups (Guilbert and
Gontard 1995).
Pure rice starch-based films exhibited the greatest TS and
E at break (Table 2). The fact that film with just starch was
founded to have the greatest TS, suggests that intermolecu-
lar forces between the starch chains were the responsible of
the film strength.
The films containing higher amount of glutelin exhibited
more rigid film. With the addition of more glutelin, film
strength decreased and so did the brittleness. The addition
of glutelin into the film formulation reduced flexibility and
increased stiffness. It indicated that rice starch-based films
containing glutelin were stiffer that related to molecular
entanglement and interactions (Zhong and Xia 2008; Su
et al. 2010). The denaturation of the protein promotes the
interaction between protein chains by intermolecular
disulphide and hydrogen bonds, resulting in a rigid matrix
with low E (Mchugh et al. 1994; Mariani et al. 2009; Oses
et al. 2009). Protein has a tendency to keep its rigid and
strong structure, being made of disulfide and hydrogen
bond (Mariani et al. 2009). The same trend was found in
the 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 2. THICKNESS, TENSILE STRENGTH (TS), % ELONGATION AT BREAK (%E) AND ELASTIC MODULUS (EM) OF COMPOSITE FILMS BASED ON
RICE 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).

Journal of Food Processing and Preservation 38 (2014) 1799–1806 © 2013 Wiley Periodicals, Inc. 1803
COMPOSITE FILMS BASED ON GLUTELIN AND RICE STARCH
FIG. 2. WATER VAPOR PERMEABILITY (WVP) VALUES OF COMPOSITE
FILMS BASED ON RICE STARCH AND GLUTELIN AT VARIOUS
AMOUNTS
number of hydrophilic interactions in the starch/glutelin
films enhances water diffusion through the film (Su et al.
2010). Polysaccharide and/or protein components of edible
films are characterized by an emphatic hydrophilicity, which
is responsible for film-water interactions and structural
changes in the polymeric film network (Kristo et al. 2007).
The resistant to water vapor decreased, with increasing
glutelin concentration. This may relate to the aggregation of
protein that causes less compact structure. The addition of
glutelin in the rice starch/glutelin composite films at high
concentration may decrease the intermolecular interactions
and increase the electrostatic repulsion between starch and
glutelin, thus increasing WVP. The results were similar to
some composite films i.e., rice protein isolate/pullulan (Shih
1996); SPI/konjac glucomanan/chitosan films (Bourtoom
and Chinnan 2008).
CONCLUSION
The incorporation of glutelin on starch films had a signifi-
cant effect on the mechanical properties with less effect on
its WVP. For films with about a ratio of glutelin and rice
starch at 25:75, improvements could be achieved with
acceptable film strength and practically no sacrifice in water
vapor resistance.
ACKNOWLEDGMENTS
This research work was supported by Research and Devel-
opment Institute, Silpakorn University, Thailand.
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