International Journal of Biological Macromolecules 67 (2014) 458462

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International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac

Preparation and characterization of bionanocomposite lms based on

potato starch/halloysite nanoclay
Fatemeh Sadegh-Hassani, Abdorreza Mohammadi Nafchi
Biopolymer Research Group, Food Science and Technology Division, Agriculture Department, Damghan Branch, Islamic Azad University, Damghan,
Semanan, Iran

a r t i c l e i n f o a b s t r a c t

Article history: In this research casting method was used to prepare potato starch based bio-nanocomposite lms with
Received 7 March 2014 halloysite nanoclay as the reinforcing materials. The composition included potato starch with 40% (w/w)
Received in revised form 30 March 2014 of a mixture of sorbitol/glycerol (weight ratio of 3 to 1as plasticizer) with nanoclay (05% w/w). The lms
Accepted 5 April 2014
were dried under controlled conditions. Physicochemical properties such as solubility in water, water
Available online 18 April 2014
absorption capacity (WAC), water vapour permeability (WVP), oxygen permeability, and mechanical
properties of the lms were measured. Results showed that by increasing the concentration of nanoclay,
Keywords:
mechanical properties of lms were improved. Tensile strength was increased from 7.33 to 9.82 MPa, and
Halloysite nanoclay, Starch lm
Mechanical properties
elongation at break decreased from 68.0 to 44.0%. Solubility in water decreased from 35 to 23%, and heat
seal strength increased from 375 to 580 N/m. Also incorporation of clay nanoparticles in the structure of
biopolymer decreased permeability of the gaseous molecules. In summary, addition of halloysite nan-
oclay, improve the barrier and mechanical properties of potato starch lms and this bionanocomposites
have high potential to be used for food packaging purposes.
2014 Elsevier B.V. All rights reserved.

1. Introduction
Starch is found in abundance in nature and due to the low price,
biodegradability, good mechanical and barrier properties, is one
of the raw materials used for food packaging. Biopolymers exhibit
high potential to be used as an alternative to plastics or synthetic
polymers as they are more environmentally friendly and are easily
biodegradable. The growth of biological products and the devel-
opment of innovative technologies, reduce dependence on fossil
fuels [1]. Sustainable and biodegradable packages are being devel-
oped worldwide and make great efforts to produce the packaging
material of natural origin (proteins, fats and carbohydrates) to take
lms or coatings. Biopolymers have advantages over the synthetic
polymers; biopolymers are biodegradable and renewable materi-
als. Permeability inhibition of gases by biopolymers increases shelf
life of fresh products such as fruits and vegetables [2]. Inhibitory
to volatile compounds and oils maintain the quality of the food
in production and distribution cycle. Besides, the lm additives in
the form of plasticizers (glycerol or sorbitol) used to enhance the
Corresponding author. Tel.: +98 232 522 5045/+98 935 501 1872;
fax: +98 232 522 5039.
E-mail addresses: fstiau@damghaniau.ac.ir,
fstiau@gmail.com (A. Mohammadi Nafchi).
exibility of biopolymer lms are also reported to impair the barrier
properties [3].
However, poor mechanical properties and high permeability
to water vapour are two main disadvantages of biopolymers that
recently nanotechnology helps to solve these problems. The use
of nanotechnology in the eld of polymer science has led to the
production of nanocomposites polymers [4]. Bionanocomposites
represent the new generation of nanocomposites, and comprises
of the combination of biopolymers and an inorganic material that
has at least one nanometer scale dimension. Bionanocomposites
is an emerging group of nanostructured hybrid materials in the
frontier between nanotechnology, material science, and life science
[5,6]. Efforts have been devoted to the development of nanobio-
composites with enhanced thermal, mechanical, and functional
properties due to the matrix or nanoparticle llers [7,8]. In addi-
tion, biopolymer-based materials are known as a green technology
and have shown biodegradability and biocompatibility in pharma-
ceutical, food packaging, and agriculture technologies [9,10].
Starch lms stronger than other biopolymer lms and mechan-
ical properties of the starch lms are better than polysaccharides
and proteins lm [11,12]. Mechanical properties of starch lms are
inuenced by many factors such as amylose to amylopectin ratio
in starch that plays an important role in the mechanical properties
of the lms. Starch lms are transparent, semi-transparent, taste-
less, colourless and avourless. Starch because of its nature, cost

http://dx.doi.org/10.1016/j.ijbiomac.2014.04.009
0141-8130/ 2014 Elsevier B.V. All rights reserved.
 F. Sadegh-Hassani, A. Mohammadi Nafchi / International Journal of Biological Macromolecules 67 (2014) 458462
and reliability of thermo plastics has a great interest as packaging
materials. However, starch alone has some disadvantages such as
hydrophilic nature and poor mechanical properties [13].
Potato starch has some number of desirable features to improve
and enhance the stability and rigidity of the gel in some foods, and
as a major component prefer to corn, wheat and other starches. The
amount of phosphorus present in the amylopectin of potato starch
has negative charge and repulsive little faster because there may be
swelling of potato starch granules in hot water and high viscosity,
high transparency and it is involved in low retrogradation speed of
potato starch pastes [14].
Incorporation of nanoparticles such as nanosilver, titanium
dioxide, nanoclay, nano zinc oxide, and silicon dioxide into food
packaging materials is reported to exhibit improved mechanical,
barrier, thermal, and physicochemical properties compared to the
pure polymers or conventional composites [10,1518]. A compos-
ite of starch and another additive might be a solution to improve
these properties. Nano clay and nano silicon dioxide are examples
of additive that is widely available, cost effective, and biodegradable
and has been shown to improve the properties of various polymer
materials [18].
Based on these observations, the major objective to undertake
the present study was to develop a biodegradable starch-based
nanocomposite lm using halloysite nanoclay to enhance their
functional properties. The method used in this research is casting
method comprising the steps of gelatinization, pasting, bursting of
granules, and starch retrogradation. It is envisaged that this type
of lms developed might nd applicability and can be exclusively
used in food packaging industry in the near future.
2. Materials and methods
2.1. Materials
Starch from potato was purchased from Glucosan Company
(Ghazvin, Iran). Glycerol and sorbitol was purchased from R&M
Marketing (Essex, UK). Halloysite nanoclay (30 0.254 m, nano-
tube) was obtained from Sigma Chemical Co (St. Louis, MO, USA).
2.2. Film preparation
Halloysite nanoclay was dispersed in water at different con-
centration (1, 2, 3, and 5% of total starch solid) and heated to
60 C with continuous stirring for 1 h, then sonicated in an ultra-
sonic bath (Marconi modelUnique USC 45 kHz) for 45 min to ensure
homogenization was completed. The solution was cooled to room
temperature and was used to prepare the aqueous starch disper-
sion 3.5% (w/w) [4]. The starch dispersion was heated to 85 C and
held at this temperature for 45 min to complete the gelatiniza-
tion. A 3:1 mixture of sorbitolglycerol at 40% (w/w of starch) was
added as plasticizers; the choice of the plasticizers was based on
the previous research by Abdorreza et al. [3]. Upon completion of
starch gelatinization, the solution was cooled to room temperature.
A portion (90 g) of the dispersion was cast on Perspex plates t-
ted with rims around the edge to yield a 16 16 cm2 lm-forming
area. Films were dried under controlled conditions in a humidity
chamber (25 C and 50% RH). Control lms were prepared similarly
but without addition of nanoparticles. Dried lms were peeled and
stored at 25 2 C and 55 5% relative humidity (RH) until experi-
mentation. All lms (including control) were prepared in duplicate.
Film thickness was measured using a micrometer (Mitutoyo Co.,
Tokyo, Japan). Overall, 10 thickness measurements were taken for
each lm and the average values were recorded.
459
2.3. Characterizations
2.3.1. Mechanical properties
ASTM D882-10 [19] was used to determine the mechanical
properties under standard conditions. Film strips were cut to
150 mm long and 25 mm wide and conditioned for 48 h in 25 C
and 55% relative humidity. Texture analyzer (TA.XT2, Stable Micro
System, Surrey, UK) equipped with Texture Exponent 32 soft-
ware was used for measuring mechanical properties of the lms.
The initial grip separation and crosshead speed were 100 mm and
0.5 mm/s, respectively. Youngs modulus, elongation at break and
tensile strength were calculated from the deformation and force
data recorded by the software. Eight replicates for every sample
were evaluated.
2.3.2. Heat seal strength measurement
Heat sealability was determined with ASTM F88/F88M-09 [20]
standard and with some modications described by Mohammadi
Nafchi et al. [21]. For each type of lm, seal strength experi-
ments were replicated ve times in a randomized, complete block
experiment. Film samples were cut into 7.62 2.5 cm strips. Two
lm strips were placed on top of one another, and an area of
(25.4 20) mm2 (at the edge of the lm) was heat-sealed using
an impulse heat-sealer Model-SP-300H (Triumph Mercantile Corp,
Taipei, Taiwan) at a temperature of 120 5 C, a sealing time of
0.9 s and heat seal pressure approximately 1.8 105 Pa. Prior to
determining seal strength all sealed lm samples were conditioned
at 23 2 C and 50 5% RH for 48 h with magnesium nitrate sat-
urated solution. The distance between the clamps was 2.5 cm. A
30-kg static load cell and a test speed of 90 mm/min were used.
Seal strength was calculated as follows:
peak force
Seal strength = (1)
lm width
The maximum force required to cause seal failure was reported
as the seal strength in Newton/metre (N/m) [20].
2.3.3. Solubility in water
Solubility of the lms in water was determined according to
Nafchi et al. [17]. Pieces of lm (2 3 cm 500 mg) were cut from
each lm and were stored in a desiccators with P2 O5 (0% RH) for 2
days. Samples were weighed to the nearest 0.0001 g and placed into
beakers with 80 mL deionized water (18 M). The samples were
stirred with constant agitation for 1 h at room temperature. The
remaining pieces of lm after soaking were ltered through lter
paper (Whatman no.1), followed by oven drying at 60 C to constant
weight. Samples were measured in triplicates. Water solubility was
calculated as following equation:
(Initial dried weight of lm Final dried weight of lm)
Solubility(%) = 100 (2)
Initial dried weight of lm
2.3.4. Water absorption capacity
The water absorption capacity measured as adapted method by
Kiatkamjornwong et al. [22]. Nanobiocomposite lms rst were
dried in a P2 O5 for one week and then a piece of 2 2 cm2 of
dried lms was added to 100 mL of distilled water and allowed to
stand for 30 min for swelling. The swollen lms were drained were
weighted. The amount of water retained by lms per dried weight
of the lms calculated as water absorption capacity.
2.3.5. Water vapour permeability (WVP)
The modied gravimetric cup method [23] based on ASTM E96-
05 [24] was used to determine the water vapour permeability
(WVP) of lms. The test cups were lled with water 1.5 cm below
the lm. A plot of weight gained versus time was used to determine
the WVTR. The slope of the linear portion of this plot represented
460 F. Sadegh-Hassani, A. Mohammadi Nafchi / International Journal of Biological Macromolecules 67 (2014) 458462
Table 1
Mechanical properties of potato starch nanocomposite lms.
Nanoclay (%) Tensile strength (MPa)
0 7.33 0.34c
1 8.09 0.49b
2 8.27 0.32b
3 8.45 0.25b
5 9.82 0.42a
Values are mean (n = 12) SD. Different letters in each column represent signicant difference at 5% level of probability among potato starch lms.
the steady state amount of water vapour transmission through the
lm per unit time (g/h). Six samples per treatment were tested. The
slopes yielded regression coefcients of 0.99 or greater. The WVP
of lm was calculated by multiplying the steady WVTR by the lm
thickness and dividing that by the water vapour pressure difference
across the lm.
2.3.6. Oxygen permeability (OP)
Oxygen permeability measurements were performed on lms
with Mocon Oxtran 2/21 (Minneapolis, USA) equipped with a
patented colometric sensor (Coulox ) and WinPermTM permeabil-
ity software. The meaurements were done using the ASTM standard
method D3985-05. The lms were placed on an aluminum foil mask
with an open area of 5 cm2 and were mounted in diffusion cells.
Tests were carried out at 25 C temperature, atmospheric pres-
sure, and 50% RH using 21% oxygen as test gas. Transferred oxygen
through the lms was conducted by the carrier (N2 /H2 ) gas to the
colometric sensor. Measurements were carried out on convergent
by hour mode to reach the steady state of oxygen transmission.
The permeability coefcients in cm3 m/(m2 day atm) were calcu-
lated on the basis of oxygen transmission rate in steady state taking
into account the lms thickness.
2.3.7. Hydrophobicity measurements
To determine the hydrophobicity of the bionanocomposite
lms, water contact-angle measurements were performed on a
static contact-angle meter (CAM-PLUS, Tantec, Germany). Data
presented are the mean of eight independent determinations at
different sites. Contact angles on each lm surface were measured
immediately after the addition of 1 L deionized water using the
Sessile Drop Half-AngleTM Tangent line method [17].
2.3.8. Color measurement
Using a colorimeter (Minolta CM-3500D; Minolta Co. Ltd., Osaka,
Japan), we studied both transmission and reection colorimetry of
triplicate lm samples. The instrument was calibrated with a zero
transmittance calibration plate CM-A100 and air as full transmit-
tance prior to use. A large size aperture was used. Color parameters
(L* , a* , b* , C* , hab ) corresponding to the uniform color space CIELAB
were obtained via the computerized system using Spectra Magic
software version 2.11 (Minolta Cyberchrom Inc., Osaka, Japan).
2.4. Statistical analysis
ANOVA and Tukeys Post Hoc tests were used to compare means
of physical, mechanical, thermal, and barrier properties of starch
based lms at the 5% signicance level. Statistical analysis was
conducted using IBMSPSS 21.0 for windows and GraphPad Prism
6 (GraphPad Software Inc., 2236 Avenida de la Playa, La Jolla, CA
92037, USA).
Elongation at break (%) Young modulus (MPa)
68.0 1.8a 188 14d
61.5 2.2b 297 5c
54.5 6.1c 315 7b
52.4 3.5c 332 14b
44.0 3.9d 376 12a
3. Results and discussions
3.1. Film formation and thickness
In the present study, control potato starch lms prepared
were exible, free standing, and transparent, while the free-
standing potato starch/halloysite nanoclay nanocomposite lms
were milky, and opaque in appearance. An increase in the milky and
opaque appearance was observed with a corresponding increase
in the content of halloysite nanoclay. The mean thickness of the
resulting nanocomposite lms remained unchanged with the addi-
tion of nanoparticles. The average values of lm thickness were
0.100.11 mm.
3.2. Mechanical properties
Tensile strength (TS), elongation at break (E), and Youngs mod-
ulus (YM) of the bionanocomposite lms are given in Table 1.
As expected from other studies on biopolymer lms reinforced
by nanoparticles [2,25], signicant increase was observed in TS
and YM and a signicant decrease was observed in elongation at
break by addition of nanoclay. Increase in TS is of vital criteria
for food packaging applications as high TS can allow the packag-
ing lms to withstand the normal stress encountered during food
handling, shipping, and transportation. The main reason is related
to the interfacial interaction between nanoclay and biopolymer
matrix. The elasticity of the lms is related to interactions of the
macromolecules and can be reduced by addition of plasticizer. For
elongation at break (E), a decreasing trend was found. Decrease
in E can be desirable and advantageous for food packaging as it is
directly related to the biodegradability of the lms [18]. As shown in
the previous researches nanoparticles signicantly reduced mois-
ture content [2]. It is likely that incorporation of nanoparticles
occupy the sites on starch that normally would be occupied by
water [25,26]. Mechanical properties of the composite lms have
been reported to be highly dependent on the interfacial interac-
tion between the matrix and llers [9]. Wu and others [27] showed
that nanoparticles, used as a lling agent, improve the wear per-
formance and tensile strength of starch lms. The nanoparticles are
likely to bond with hydroxyl groups and other possible hydrogen
or Van der Walls bonds of starch macromolecules strengthening
molecular forces between nanoparticles and starch [27].
3.3. Heat seal strength
Heat sealability is a critical factor in use of lms for packaging
purposes. Fig. 1 shows the effect of nanoclay on heat sealability of
potato starch lms. Potato starch lms without nanoparticles had
good heat sealability and the seal strength of near 500 N/m which is
comparable with that of synthetic lms (>600 N/m). Introduction of
nanoclay improved heat sealability. These results support the the-
ory proposed by Kim and Ustunol [28] using electron spectroscopy
for chemical analysis (ESCA). They reported that increasing hydro-
gen and covalent bonds involving C O H and N C, which may
be the main forces responsible for the sealed joint formation of
 F. Sadegh-Hassani, A. Mohammadi Nafchi / International Journal of Biological Macromolecules 67 (2014) 458462 461

Fig. 1. Effects of nanoclay on seal strength of potato starch lms. Values are mean Fig. 3. Water absorption capacity of potato starch lms and effects of nanclay on
(n = 8) SD. Bars with different letters are signicantly different at 5% level of prob- WAC of lms in water at 25 C. Values are means (n = 5) SD and bars with different
ability. letters are signicantly different at 5% level of probability.

the lms. It was likely that Si O bonds interconnecting starch components [32]. For this reason, free water molecules do not inter-
molecules improved the heat sealability. These results are consis- act as strongly as with nanocomposite lms as with composite lms
tent with data reported by Abdorreza and Karim [21]. alone. These results not only consistent with the water solubil-
ity results but also with results reported by other researchers on
nanobiocomposites [2,32,33].
3.4. Solubility in water and water absorption capacity
3.5. Hydrophobic effects of halloysite nanoclay
Solubility in water may be an important factor in dening appli-
cations for biopolymer composite lms. Most of the biopolymers
One method of comparing the hydrophobicity of material sur-
are sensitive to water. By incorporating lipids, crosslinking of the
faces is the determination of the contact angle of a water droplet on
structure or incorporation of nanoparticles sensitivity to water
the surfaces. A high contact angle indicates greater hydrophobic-
could be decreased [29]. Fig. 2 shows the solubility of potato starch
ity of the surface and vice versa. The contact angle increased with
lms in deionized water after 1 h.
increasing halloysite nanoclay content of potato starch lms (Fig. 4),
Incorporation of nanoparticles suppressed the diffusion of water
which indicates that the tendency to absorb water decreased and
into the structure. Effects of nanoclay on the water resistance of
that the surfaces became more hydrophobic. These ndings concur
biopolymeric lms reported by other researchers and these results
with the ndings of water solubility and water absorption capacity
are consistent with their results [18,27,30,31].
investigations detailed above.
Water absorption capacity of the nanoclay biocomposite lms
are given in Fig. 3. Introducing nanoclay to potato starch matrix
3.6. Water vapour permeability
signicantly decrease the water absorption capacity of the potato
starch. This could be attributed to the interactions between clay
Decrease in the water vapour permeability is of high importance
and starch in lm structure. The experimental results showed that
when developing food packaging materials where efcient barrier
when the nanoparticle content of potato starch lms was increased,
properties are desired to minimize moisture transfer between out-
more hydrogen bonds formed between the SiO2 and the matrix
side packaging environment and packaged food. Table 2 shows
the effect of nanoparticles on the water vapour transmission
rate through potato starch lms. As expected, introduction of

Fig. 2. Water solubility of potato starch lms and effects of nanclay on solubility of Fig. 4. Water contact angle on potato starch lms surfaces as a function of nanoclay
lms in water at 25 C. Values are means (n = 5) SD and bars with different letters content. The bars show mean (n = 8) SD. Different letters on the bars represent the
are signicantly different at 5% level of probability. signicant difference at 5% level of probability.
462 F. Sadegh-Hassani, A. Mohammadi Nafchi / International Journal of Biological Macromolecules 67 (2014) 458462
Table 2
Water vapour permeability, oxygen permeability, and tortuosity value of potato starch nanocomposites.
Nanoclay (%) WVP 1011 [g/m/s/Pa]
0 2.91 0.53a
1 2.14 0.29b
2 1.44 0.24c
3 1.02 0.12d
5 0.83 0.11d
Values are mean (n = 5) SD. Different letters in each column represent signicant difference at 5% level of probability among potato starch lms. O.P.: Oxygen permeability
coefcients at 25 C and 50% RH. : tortuosity value from Nielsons equation.
nanoparticles signicantly decreased water vapour permeability
(WVP) of potato starch lms. Nanoparticles lling the pores in the
macromolecules structures can decrease gas permeability as well
as permeability to water vapour [34]. Also other researchers have
shown positive effects of nanoparticles on WVP. For example, Baj-
pai et al. [30] have shown that WVP of chitosan lms added with
zinc oxide nanoparticles decreased signicantly.
3.7. Permeability to oxygen
The results of oxygen permeability are presented in Table 2. Oxy-
gen permeability coefcient signicantly decreased from 70 to
45 cm3 m/(m2 day atm) by addition of halloysite nanoclay. The
inorganic nanoclay particles are more water resistance than the
biocomposite matrix; therefore incorporation of these nanoparti-
cles to the matrix could likely introduced a tortuous pathway for
oxygen molecules to pass through [35]. Based on the Nielsens [36]
simple model on tortuosity, it is possible to describe the permeabil-
ity reduction in halloysite nanoclay incorporated biocomposites.
The reduction in permeability coefcients means that gases should
travel in a longer diffusive path. The relation between tortuosity
and permeability as given by Nielsens [36] model is
pc 1 s
= (3)
pm 
where  is tortuosity, s is the volume fraction of halloysite
nanoclay, Pm and Pc are the permeability coefcients of the bio-
composite and bionanocomposite, respectively. Table 2 shows the
tortuosity value for each bionanocomposite matrix. Zeppa et al.
[37] and Nafchi et al. [2] also reported similar results for oxygen
permeability by incorporation of nanoparticles.
3.8. Colour characteristics
The color characteristic parameters for potato starch and
nanoclay-incorporated potato starch nanocomposites are reported
in Table 3. Overall, depending on the amount of nanoparticles
added, the resulting lms exhibited an increase in brightness and
different color values. The lightness (L* ) signicantly decreased
with increasing halloysite nanoclay content. The transmissions of
the lms in different wavelengths signicantly decreased upon
addition of nanoparticles. This means that addition of nanoparticles
to the lms will improve the ability of the lms to protect package
contents from light and will enhance the quality of the packaged
Table 3
Transparent colorimetric parameters of the starch lms.
Nanoclay (%) L* a*
0 94.46 0.71a 0.14 0.07d
1 91.18 0.41b 0.48 0.23c
2 87.04 0.40c 0.75 0.16b
3 79.09 0.24d 0.96 0.37b
5 64.30 0.23e 1.70 0.08a
Values are mean (n = 5) SD. Different letters in each column represent signicant
difference at 5% level of probability among potato starch lms.
O.P. [cm3 /m/(m2 day)] O.P.composite /O.P.matrix 
76.88 2.44a 1 1
61.72 1.28b 0.80 1.24
50.98 1.02c 0.66 1.50
45.77 0.50d 0.59 1.67
38.20 0.81e 0.50 1.99
food. The same results were reported for the effects of nano SiO2
on potato starch nanocomposites [21].
4. Conclusion
In this study, we introduced halloysite nanoclay to potato
starch matrix to fabricate bionanocomposites. Introduction of the
nanoparticles improved mechanical properties of the lms made
from potato starches. Permeability to water vapour, and oxygen,
water absorption capacity, and water solubility of the lms signif-
icantly decreased. The results showed that under strict regulation,
bionanocomposites based on halloysite nanoclay may have poten-
tial applications in food packaging industries.
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