Food Hydrocolloids 44 (2015) 172e182

Contents lists available at ScienceDirect

Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Fabrication of bio-nanocomposite lms based on sh gelatin

reinforced with chitosan nanoparticles
Seyed Fakhreddin Hosseini a, *, Masoud Rezaei a, Mojgan Zandi b, Farhid Farahmandghavi c
a

Department of Seafood Processing, Faculty of Marine Sciences, Tarbiat Modares University, P.O. Box 46414-356, Noor, Iran
Department of Biomaterials, Iran Polymer and Petrochemical Institute, P.O. Box 14965/115, Tehran, Iran
c
Department of Novel Drug Delivery Systems, Iran Polymer and Petrochemical Institute, P.O. Box 14965/115, Tehran, Iran
b

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 25 December 2013
Accepted 1 September 2014
Available online 28 September 2014

The paper focuses on the synthesis of chitosan nanoparticles (CSNPs) by ionic gelation between chitosan
(CS) and sodium tripolyphosphate (TPP) and subsequently its use as ller in a sh gelatin (FG) matrix to
produce bio-nanocomposite lms. The obtained particles exhibited a spherical shape with size range of
40e80 nm, and a positively charged surface with a zeta potential value of 10 mV. XRD results conrmed
the cross-linking reaction between CS and TPP. SEM images showed that CSNPs could be well dispersed
in FG polymer matrix at low content, while higher CSNPs loadings (8%, w/w) resulted in the aggregation
of particles in the composites. FTIR spectroscopy results conrmed the interaction between CSNPs and
FG through hydrogen bonding. The nucleating effect of the CSNPs was conrmed by DSC analysis. Results
indicated that the addition of CSNPs caused remarkable increase in the tensile strength (TS) and elastic
modulus (EM), which leading to stronger lms as compared with individual FG lms, but decreased the
elongation at break (EAB). Furthermore, addition of CSNPs contributed to the signicant decrease
(p < 0.05) of water vapor permeability (WVP), leading to a 50% decline at 6% (w/w) ller. The light barrier
measurements presented low values of transparency at 600 nm of the FG-based nanocomposite lms,
indicating that these lms are very transparent (lower in transparency value) while they have excellent
barrier properties against UV light. The results presented in this study show the feasibility of using bionanocomposite technology to improve the properties of biopolymer lms based on FG.
2014 Elsevier Ltd. All rights reserved.

Keywords:
Bio-nanocomposite
Edible lms
Fish gelatin
Chitosan nanoparticles
Characterization

1. Introduction
Increasing and widespread environmental awareness, as well as
efforts to reduce the volume ow of wastes and increase the use of
renewable raw materials have placed emphasis on the disposal
properties of different materials (Endres & Siebert-Raths, 2012).
The non-degradable and non-renewable nature of plastic packaging has led to a renewed interest in packaging materials based on
biopolymers derived from renewable sources. The use of
biopolymer-based packaging materials can solve the waste disposal
problem to a certain extent (Kumar, Sandeep, Alavi, Truong, &
Gorga, 2010). The increscent interest in biopolymer based packaging has resulted in the development of protein-based lms from
soy protein, whey protein, casein, collagen, corn zein, gelatin, and
wheat gluten (Cuq, Gontard, & Guilbert, 1998). Among all the

* Corresponding author. Tel.: 98 122 625 3101x3; fax: 98 122 625 3499.
E-mail
addresses:
[email protected],
[email protected]
(S.F. Hosseini).
http://dx.doi.org/10.1016/j.foodhyd.2014.09.004
0268-005X/ 2014 Elsevier Ltd. All rights reserved.

protein sources, gelatin has also been extensively studied for its
lm forming capacity and applicability as an outer covering to

mez-Guille

n et al.,
protect food against drying, light and oxygen (Go
2009).
Fish gelatin (FG) has gained great interest in recent years as the
demand for non-bovine and non-porcine gelatin has increased, due
to religious and social reasons, and also the bovine spongiform
encephalopathy (BSE) crisis (Bae et al., 2009). Furthermore, sh
skin, which is a major byproduct of the sh-processing industry,
causing waste and environmental pollution, could provide a valuable source of gelatin (Badii & Howell, 2006). The elaboration of

mezedible lms from sh gelatin has been recently studied (Go

mez-Guille

n, Ferna

ndez-Martn, & Montero, 2011;
Estaca, Go
~ ez-Flores
Hosseini, Rezaei, Zandi, & Farahmandghavi, 2013; Nn
et al., 2012; Nur Hanani, Roos, & Kerry, 2012).
However, these biodegradable sh gelatin lms do have some
limitations in their use, such as low tensile strength (TS) and high

mez-Estaca et al., 2011). In order to improve the
water solubility (Go
mechanical property as well as barrier characteristic of gelatin lms,
recently, a new class of materials namely bio-nanocomposites

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

(biopolymer matrix reinforced with nanoparticles) has introduced as

a promising option (Bae et al., 2009). Fillers with at least one nanosized dimension (nanollers or nanoreinforcements) have better
interfacial adhesion with the polymer matrices, when compared to
the respective micro/macroscopic reinforcements. A uniform
dispersion of nanollers leads to a very large matrix/ller interfacial
area, changing the molecular mobility, improve the relaxation
behavior, and the consequent thermal and mechanical properties of
~ a, Alvarez, & Vasquez, 2007).
the resulting nanocomposite (Luduen
Nanocomposite technology using nanollers such as carbon nanotubes (CNTs) (Ma, Yu, & Wang, 2008), nanoclay (Bae et al., 2009;
Casariego et al., 2009), and nanosilica (Ahmed, Varshney, & Auras,
2010) has already proved to be an effective way to improve the
mechanical, physical, and thermal properties of polymers. Newly,
considering of the applications for edible lms and/or food packing,
much attention has been focused on polysaccharide nanollers.
Chitosan (CS) is a naturally occurring nontoxic, biocompatible,
biodegradable, and cationic polysaccharide (Shahidi, Arachchi, &
Jeon, 1999).
Chitosan nanoparticles (CSNPs) which are composed of a natural
material with excellent physicochemical properties, is environmentally friendly, and bioactive (Yang, Wang, Huang, & Hon, 2010).
CSNPs can be prepared by the electrostatic interaction and resultant ionotropic gelation between CS polycation and sodium tripo~ na

n-Lo

pez, & Vila-Jato
lyphosphate (TPP) polyanion (Calvo, Remu
Alonso, 1997; Yang et al., 2010). Using of these nanoparticles in
edible lms would be very promising, due to the food-grade
properties of both components. De Moura et al. (2009) found that
CS-TPP nanoparticles increases thermal and mechanical properties
and decrease water vapor permeability of the hydroxypropyl
methylcellulose (HPMC) lms. In another study, these polysaccharide nanoparticles have been used as the reinforcing medium in glycerol plasticised-starch (GPS) matrices (Chang, Jian, Yu,
& Ma, 2010). They have obtained an improvement of thermal stability, mechanical and barrier properties of GPS composites. More
recently, Martelli, Barros, De Moura, Mattoso, and Assis (2013) was
also reported that the incorporation of chitosan nanoparticles
promoted noticeable improvement of the mechanical properties
and acted in reducing the water vapor permeation rate in banana
puree lms.
Hence, bio-nanocomposite lms based on FG and CSNPs could
be a good candidate for food packaging applications to extent the
shelf life of foods and products. This research focused on fabrication
and characterization of CSNPs as well as evaluation of the effects of
incorporation of obtained nanoparticles on morphology, mechanical properties, water vapor permeability, light barrier properties,
and thermal behavior of FG lms.
2. Materials and methods
2.1. Materials
Gelatin from cold water sh skin, chitosan (CS) (medium molecular weight, 75e85% deacetylated) and sodium tripolyphosphate (TPP) were purchased from SigmaeAldrich (St. Louis, MO,
USA). Glycerol (analytical grade) and acetic acid were purchased
from Merck Chemicals Co., (Darmstadt, Germany).
2.2. Preparation of chitosan nanoparticles (CSNPs)
CSNPs were prepared based on the ionotropic gelation between
CS and TPP with some modications (Calvo et al., 1997). CS solution
(1% (w/v)) was prepared by agitating chitosan in an aqueous acetic
acid solution (1% (v/v)) at ambient temperature (23e25  C) overnight. The mixture was then centrifuged at 9000 rpm for 30 min;

173

the supernatant was separated and ltered through 1 mm pore size

lters. TPP solution (0.4% (w/v)) was added (weight ratios of CS:
TPP was 2.5:1) drop wise to CS solution under vigorous stirring for
40 min. The formed particles were collected by centrifugation at
9000 rpm for 30 min at 4  C, and subsequently washed several
times with deionized water. Finally, ultrasonication (50 w) was
applied by a sonicatior (Bandelin sonopuls HD3200, KE 76 probe,
Germany) in an ice bath for 4 min with a sequence of 0.7 s of
sonication and 0.3 s of rest, resulting in a homogeneous suspension.
The suspensions were immediately lyophilized at 35  C for 72 h
using freeze dryer (GAMMA 1-16 LSC, UK).
2.3. Bio-nanocomposite lm preparation
The gelatin lms were prepared according to the method

mez-Estaca et al. (2011) with slight modications.
described by Go
The FG solution was prepared by dissolving 4 g gelatin in 100 ml
distilled water for 30 min and then heated at 45  C for 30 min under
continuous stirring. Glycerol (0.3 g/g gelatin) was added as a
plasticizer and solutions were again warmed and stirred at 45  C for
15 min. For the preparation of bio-nanocomposite lm forming
dispersions, different levels of CSNPs (0, 2, 4, 6 and 8%, w/w), based
on dry FG was rst dispersed into distilled water (50 ml) and then
sonicated for 10 min. Then, the CSNPs suspension were added to FG
solution (50 ml) drop wise and gently stirred for 60 min. The lmforming dispersions were degassed under vacuum for 15 min to
remove air bubbles. Finally, aliquots of 100 g of lm-forming dispersions were poured in rectangular plastic dishes (24  12 cm) and
dried at ambient temperature (23e25  C) for 3 days. Dried lms
were peeled from the plate and conditioned at 25  C and a relative
humidity (RH) of 50 4% RH for 48 h for further analysis.
2.4. Characterization of CSNPs and bio-nanocomposite lms
2.4.1. Atomic force microscopy (AFM)
Atomic force microscope (DualScopeTM DS95-50, DME,
Denmark) was used for morphological characterization and nanoparticles size determination. A drop of diluted nanoparticle suspension (0.05 mg/ml) was spread on the clean glass surface, and
dried at room temperature. The image measurement was performed in tapping mode using silicon probe cantilever of 230
micron length, resonance frequency of 150e190 kHz, spring constant of 20e60 N/m and nominal, 5e10 nm tip radius of curvature.
The scan rate was used as 1 Hz.
2.4.2. Particle size and zeta potential
The mean particle size, particle size distribution and zeta potential of the nanoparticle suspension were obtained using a
Zetasizer Nano ZS 3300 (Malvern Instruments Ltd., United
Kingdom) on the basis of dynamic light scattering (DLS) technique.
Diluted samples were placed in glass cuvette with square aperture
and the scatter intensity was measured at 25  C.
2.4.3. X-ray diffraction
XRD patterns of CS powder and CSNPs were recorded over a 2q
range of 5e50 using an X-ray diffractometer (Siemens, model
D5000) with a step angle of 0.04 /min.
2.4.4. Fourier transform infrared (FTIR) spectroscopy
FTIR spectra of CS powder, CSNPs and bio-nanocomposite lms
were recorded from wavenumber 400e4000 cm1 by a Bruker
Equinox 55 spectrometer (Bruker Banner Lane, Coventry, Germany). For CS powder and CSNPs, samples were prepared using KBr
to form pellets. For each spectrum, 16 scans at a resolution of
4 cm1 were obtained.

174

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

2.4.5. Scanning electron microscopy (SEM)

The morphology of the lm samples was investigated by scanning electron microscopy (SEM) (VEGA II, TESCAN, Czech Republic)
at an accelerating voltage from 10 to 20 kV. Strips of dry lms
(obtained by maintaining in a desiccator with silica gel for 2 week)
were immersed in liquid nitrogen and cryo-fractured manually. The
specimens were stuck onto a cylindrical aluminum stub by a
double-sided tape. Afterward the lm samples were sputtered with
a thin layer of gold in an ion sputter coater (K-450X, EMITECH,
England) and placed into SEM chamber to observe the surface and
the cross-section of the lms.
2.4.6. Differential scanning calorimetry (DSC) analysis
Thermal properties of lms were determined using a differential
scanning calorimeter (DSC, Model 200-F3 Maia, Netzsch, Germany).
Before analysing, lms were conditioned in a desiccators containing dried silica gel for 2 weeks at room temperature (23e25  C) to
obtain dehydrated lms. Aliquots of approximately 10 mg pre-dried
samples were placed into aluminum pans, sealed and scanned over
the range of 25e350  C with a heating rate of 10  C/min. The empty
aluminum pan was used as a reference.
2.4.7. Film thickness
Thickness of the lms was determined using a micrometer
(Mitutoyo Manufacturing Co. Ltd., Tokyo, Japan) to the nearest
0.001 mm at 9 random positions around the lm, and average
values were used in calculations.
2.4.8. Mechanical properties
A universal testing machine (SMT-20, Santam, Tehran, Iran)
equipped with a 60 N load-cell was used to measure tensile
strength (TS), elongation at break (EAB) and elastic modulus (EM)
according to the ASTM standard method D 882-09 (2009). Films
strips of 110  20 mm were conditioned at 23 2  C and 53 2%
relative humidity for 48 h in an environmental chamber before
testing. Initial grip separation and mechanical crosshead speed
were set at 50 mm and 5 mm/min, respectively. Each type of lm
was tested by at least ve replicates.
2.4.9. Water vapor permeability (WVP)
The water vapor permeability of the lms was measured
gravimetrically according to the ASTM E96-05 method (2005) and
adapted to hydrophilic edible lms by McHugh, Avena-Bustillos,
and Krochta (1993). Circular test cups were made of glass with
49 mm internal diameter and 1.1 cm height. Films without pinholes
or any defects were sealed to the cup mouth containing 6 ml
distilled water (100% RH; 2.337  103 Pa vapor pressure at 20  C),
placed in a desiccators at 20  C and 0% RH (0 Pa water vapor
pressure) containing silica gel. The water transferred through the
lm and adsorbed by the desiccant was determined from the
weight loss of the permeation cell. The lms were allowed to
equilibrate for 2 h before the cells were initially weighed. The cells
were weighted at intervals of 2 h during 10 h with an analytical
balance (0.0001 g). The slope of weight loss versus time was obtained by a linear regression (r2  0.99). The measured WVP of the
lms was determined using Eq. (1):

WVP

WVTR  L
DP

(1)

Where WVTR is the water vapor transmission rate (g mm/kPa h m2)

through a lm, calculated from the slope of the straight line divided
by the exposed lm area (m2), L is the mean lm thickness (mm),
and DP is the partial water vapor pressure difference (kPa) across
the two sides of the lm. Three replicates of each lm were tested.

2.4.10. Water solubility

Water solubility of the lms was measured according to the
method of Gontard, Guilbert, and Cuq (1992). Three pieces
(1  4 cm2) of the lms were weighed (0.0001 g) and subsequently dried in an air-circulating oven at 105  C for 24 h. After this
time, lms were recovered and re-weighed (0.0001 g) to determine their initial dry weight (Wi). Afterward, the samples were
immersed in 30 ml of distilled water and the system gently shaken
(100 rpm) for 24 h at room temperature (22e25  C). The samples
were then passed through a weighed lter paper (Whatman 1). The
lter paper together with unsolubilized fraction was dried in a
forced-air oven (105  C, 24 h) and weighed (Wf). The lm solubility
(FS%) was calculated using Eq. (2):

FS%

Wi  Wf
 100
Wi

(2)

Wi initial dry lm weight (g)

Wf nal dry lm weight (g)

2.4.11. Light transmission and transparency

The barrier properties of the lms against ultraviolet (UV) and
visible light were measured at selected wavelength between 200
and 800 nm, using a UV-1650 spectrophotometer (Model PC, Shimadzu, Kyoto, Japan). Film strips of 10  40 mm were placed in a
spectrophotometer test cell. An empty test cell was used as the
reference. The measurement was done in triplicate and the average
of three spectra was calculated. Transparency of the lms was
calculated as in the following equation (Han & Floros, 1997):

Transparency A600 =x

(3)

where A600 is the absorbance at 600 nm, and x is the lm thickness

(mm). The greater transparency value represents the lower
transparency.
2.4.12. Statistical analysis
The statistical analysis of the data were performed using analysis of variance (ANOVA) and the differences between means were
evaluated by least signicant difference multiple range test at
p  0.05. SPSS statistic program (SPSS 17.0 for window, SPSS Inc.,
Chicago, IL, USA) was used for data analysis.
3. Results and discussion
3.1. Characterization of CSNPs
AFM imaging is an effective method to provide the surface
morphology and accurate size and size distribution of the particles.
AFM images conrmed the spherical shape and nanosize structure
of CSNPs (Fig. 1a). The size distribution obtained by AFM indicated
that most of CSNPs were distributed in the range of 40e80 nm
(Fig. 1b). However, the size of the CSNPs is smaller than those obtained by De Moura et al. (2009), using the same process but
different CS and TPP concentration, describing particles with size
range of 85e221 nm. These authors reported that the nanoparticles
with lowest particle size (85 nm) were obtained when the lowest
CS and TPP concentration was used. Chang et al. (2010) observed
when commercial CS was used to prepare CSNPs at a concentration
of 1% (w/v) of CS and TPP, the particle size distribution was in the
range of 50e100 nm. The effect of CS/TPP ratio on particle size was
previously studied. Calvo et al. (1997) reported that the particle size
is dependent on both CS and TPP concentrations.

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

175

Fig. 1. (a) AFM images (3D) and (b) size distribution obtained from the height of CSNPs.

Dynamic light scattering (DLS) technique was also applied to

investigate the mean particle size and particle size distribution of
the nanoparticles. Fig. 2a shows a typical size distribution prole of
the nanoparticles with a mean diameter of 259 17.9 nm in a
narrow size distribution (polydispersity index <1). The size
measured in hydrated state is reported to be slightly higher than
the size measured in dried state (Yoksan, Jirawutthiwongchai, &
Arpo, 2010). It should be pointed out that the particle size determined by DLS technique is a hydrodynamic diameter. The larger
diameter thus might be a result of the swelling of the chitosan layer
surrounding the individual particles, and/or the aggregation of
single particles while dispersed in water (Yoksan et al., 2010).
Particle aggregation can be prevented by electrostatic repulsions
according to DLVO theory (Derjaguin & Landau, 1993; Verwey &
Overbeek, 1948). Electrostatic repulsion between particles

depends on the value of zeta potential. The higher the zeta potential, the stronger the repulsion, the more stable the system becomes. CSNPs showed zeta potential values about 10 mV (Fig. 2b).
The positive zeta potential found on the surface of the particles is
due to the cationic characteristics of the chitosan chains (Lorevicel,
De Moura, Aouada, & Mattoso, 2012). Dudhani and Kosaraju (2010),
and Liu et al. (2011), also reported the positively charged surface of
chitosan-TPP particles. Lorevicel et al. (2012) showed that the zeta
potential of chitosan nanoparticles can vary between 14.50 and
37.58 mV, depending on chitosan concentrations used.
The XRD patterns of CS powder and CSNPs are presented in
Fig. 3. CS exhibits one characteristic peak at 2q of 25 (Fig. 3a),
indicating the high degree of crystallinity (Ali, Rajendran, & Joshi,
2011; Jingou et al., 2011). After ionic cross-linking with TPP, no
peak is found in the diffractogram of chitosan nanoparticles,

Fig. 2. (a) Size distribution and (b) zeta potential distribution of CSNPs determined by DLS technique.

176

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

electrostatic interaction between NH

3 groups of CS and polyphosphoric groups of TPP to enhance both the intermolecular and
intramolecular interaction in CSNPs (Dudhani & Kosaraju, 2010;
Jingou et al., 2011; Qi, Xu, Jiang, Hu, & Zou, 2004; Yoksan et al.,
2010). Moreover, the shoulder at 1258 cm1 reduced dramatically
or disappeared and a new peak was formed at 1238 cm1 (CeOeC
stretch).
3.2. Film microstructure

Fig. 3. XRD patterns of (a) CS powder and (b) CSNPs.

reecting the destruction of the native chitosan packing structure

(Yoksan et al., 2010) (Fig. 3b). It is well-known that the width of Xray diffraction peak is related to the size of crystallite, the broadened peak usually results from imperfect crystal (Jingou et al.,
2011). So the broad peak of CSNPs may be caused by the crosslinking reaction between CS and TPP, which may destroy the
crystalline structure of CS (Rokhade et al., 2006).
The FTIR experiments of CS and CSNPs were performed to
evaluate the chemical structure of nanoparticles. Fig. 4 shows FTIR
spectra of CS powder and CSNPs. In general, CS powder shows
characteristic peaks at 3433 (-OH and -NH2 stretching), 2920 (-CH
stretching), 1647 (amide I), 1088 (CeOeC asymmetric stretching)
and 591 cm1 (pyranoside ring stretching vibration) (Fig. 4a). The
sharp peaks at 1380 cm1 and 1419 cm1 can be assigned to CH3
symmetrical stretching band (Dudhani & Kosaraju, 2010). After
formation of CSNPs (Fig. 4b), the peak of amide I (-NH2 bending)
was shifted from 1647 to 1651 cm1, and a new peak appeared at
1555 cm1 (amide II), implying the complex formation via

Permeability of lms can be affected by the structure,

morphology, and homogeneity of the matrix (McHugh & Krochta,
1994). SEM micrographs of the surfaces of neat FG and prepared
nanocomposite lms are shown in Fig. 5. Pure FG lm displays
smooth and homogeneous surfaces, without pores and with
excellent structural integrity (Fig. 5a). Different surface
morphology was observed after CSNPs was added into FG lms. At
low CSNPs content (2%, w/w), most of CSNPs are dispersed uniformly in the FG matrix without obvious aggregation (Fig. 5b). As
illustrated in Fig. 5c, when 6% (w/w) CSNPs was added to the FG
matrix, slightly aggregation was observed, whereas at high ller
content (i.e. 8%, w/w) agglomeration of CSNPs was obvious (Fig. 5d).
Nevertheless, in comparison with neat sh gelatin lm, relatively
dense structure was observed after adding 2, 6 and 8% CSNPs. This
explained by the improved mechanical and barrier properties of
the nanocomposite lms due to good bonding strength between
nanoparticles and polymer. Similar observation was reported by
Chang et al. (2010) that worked with starch-chitosan nanoparticles
composites.
Fig. 6 represents the cross-section morphology of FG and FGbased nanocomposite lms. It is clear that addition of CSNPs
caused changes in the lm microstructure, since the non-reinforced
lms (Fig. 6a) exhibited a smooth and continuous surface, as expected for a homogeneous material. SEM micrographs obtained
from the samples containing 2% (w/w) CSNPs showed a good distribution of particles in the FG matrix, without aggregates formation (Fig. 6b). However, addition of 6% (w/w) CSNPs led to a rough
surface, with a small aggregates of particles in which is possible to
see Fig. 6c. In the case of FG lms with 8% (w/w) CSNPs, obvious
aggregation was observed in part of the lm matrix (Fig. 6d). The
similar microstructure has been reported in alginate lm incorporated with nanocrystalline cellulose (Huq et al., 2012).
3.3. FTIR analysis

Fig. 4. FTIR spectra of (a) CS powder and (b) CSNPs.

FTIR spectroscopy analysis was performed to better understand

the effects of CSNPs ller on the structure of the FG matrix, and the
results are shown in Fig. 7. The spectrum of FG lm (Fig. 7a) showed
characteristic bands at approximately 3291 cm1 (amide-A, NeH
stretching), 1679 cm1 (amide-I, C]O stretching), 1540 cm1
(amide-II, NeH bending) and 1242 cm1 (amide-III, CeN and NeH
stretching) (Pereda, Ponce, Marcovich, Ruseckaite, & Martucci,
2011). The characteristic peak appeared at 2935 cm1 corresponds to the CeH stretching vibration (Kanmani & Rhim, 2014).
The band situated at the wavenumber of ~1046 cm1 was found in
all lm samples, corresponding to the glycerol (-OH group) added
as a plasticizer (Bergo & Sobral, 2007). The FTIR peaks of FG-based
nanocomposite lms are relatively similar to those of FG lm as
control. As it could be observed, some of the peaks are shifted to
higher or lower wavenumbers with increasing CSNPs loading. For
example, the peaks of amide-A and amide-II shifted to higher
wavenumbers as shown in Fig. 7bed, indicating a possible formation of hydrogen bonding between nanoparticles and polymer
matrix (Khan et al., 2012). The shift of the amide-A band indicated
that the hydrophilicity of gelatin was reduced, because the free

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

177

Fig. 5. SEM micrographs of the surface of (a) FG and FG-based nanocomposite lms containing (b) 2%, (c) 6%, and (d) 8% (w/w) CSNPs content.

eOH group of gelatin was involved in hydrogen bonding association with CSNPs and less susceptible to hydration (Le Tien et al.,
2000). In gelatin eOH, the eCOOH end groups and eNH2, and the
side groups are capable of forming hydrogen bonds with the eOH
and the eNH2 groups of chitosan (Sionkowska, Wisniewski,
Skopinska, Kennedy, & Wess, 2004). Khan et al. (2012) reported
the fabrication of chitosan based nanocomposite lms reinforced
with varying concentrations of nanocrystalline cellulose (NCC)
(1e10%, w/w). These authors represented that the shift of amide-A
peak to higher wavenumber was associated with formation of
hydrogen bonding between chitosan and NCC. In another study, the
shift of amide-II peak in gelatin/clay composite lms to higher
wavenumber was also attributed to the strong interaction between
nanoclay and polymer matrix through the hydrogen bonding
(Kanmani & Rhim, 2014).
3.4. Thermal properties
DSC thermograms of the pure FG and fabricated nanocomposites, are shown in Fig. 8. Melting points (Tm) of the
samples were evaluated from the position of the maximum in the
endothermic peaks of DSC thermograms. As shown in Fig. 8a, FG
exhibited two sharp endothermic peaks at 243.5  C and 264.9  C,

implying Tm of the material possessing two main different crystal

structures. The crystal melted at lower temperature might be
assigned to the devitrication of blocks rich in a-amino acids;
while the one melted at higher temperature might be attributed
to the devitrication of blocks rich in imino acids such as proline
and hydroxyproline (Zheng, Li, Ma, & Yao, 2002). By addition of 2
and 4% (w/w) CSNPs, melting peaks was alleviated and shifted to
lower temperatures (Fig. 8b and c). The temperature position of
the melting peaks decreased from 239.7 to 237.5  C (rst endothermic peak) and 258.8 to 255.8  C (second endothermic peak)
as CSNPs content increased from 2 to 4% (w/w), respectively. The
lower Tm observed in the heating run can be an indication of
faster crystallization induced by CSNPs which act as nucleating
agents for FG. CSNPs allow heterogeneous nucleation mechanism
which induces a decrease of the free energy barrier and fastens
the crystallization. This observation is in agreement with the
results of Frone, Berlioz, Chailan, and Panaitescu (2013) related to
the effect of nucleating agents of cellulose nanobers on the
thermal properties of polylactic acid (PLA) composites. However,
by increasing of CSNPs contents (i.e. 6 and 8%, w/w), only a sharp
endothermic peak was observed (Fig. 8d and e), reecting that
the obtained bio-nanocomposite lms possessed sole-structured
crystals.

178

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

Fig. 6. SEM micrographs of the cross-section of (a) FG and FG-based nanocomposite lms containing (b) 2%, (c) 6%, and (d) 8% (w/w) CSNPs content.

3.5. Film thickness

Film thickness is a crucial parameter in calculation of mechanical properties and water vapor permeability values. Final thickness
strongly depends on preparation method and drying conditions.
The lm thickness increased from 50.5 2.8 mm for the control FG
lm to 64.8 3.3 mm for bio-nanocomposite lm with 8% (w/w)
CSNPs (Table 1). This behavior could be as a result of the increase in
dry matter content. Those values are lower than those presented in
the literature for nanocomposite lms based on sh gelatin and
montmorillonite (MMT) nanoclay (Jeya Shakila, Jeevithan,
Varatharajakumar, Jeyasekaran, & Sukumar, 2012). Moreover, the
values were lower than those reported by Pereda et al. (2011) for
chitosan-gelatin composite lms with thickness of 100 6 mm. This
was due to the differences in lm forming solution formulations
and in the used lm-making procedures.
3.6. Mechanical properties of lms

Fig. 7. FTIR spectra of (a) FG and FG-based nanocomposite lms containing (b) 2%, (c)
6%, and (d) 8% (w/w) CSNPs content.

It is highly desirable that an edible lm maintains its integrity

during processing, shipping, and handling. Mechanical properties
are important in edible lms, because adequate mechanical
strength ensures the integrity of the lm and its freedom from
minor defects such as pinholes (Murillo-Martnez, Pedroza-Islas,

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

179

Fig. 8. DSC thermograms of (a) FG and FG-based nanocomposite lms containing (b)
2%, (c) 4%, (d) 6% and (e) 8% (w/w) CSNPs content.

Lobato-Calleros, Martnez-Ferez, & Vernon-Carter, 2011). Representative stress-strain curves of the lms are shown in Fig. 9. These
curves exhibited the typical plastic deformation behavior; at low
strains (<10%) the stress increased rapidly with an increase in the
strain and the slopes were in the elastic region dening the elastic
modulus. At strains >10%, the stress decreased slowly until failure
occurred.
Table 1 shows the mechanical properties of the lms, in terms of
tensile strength (TS), elongation at break (EAB), and elastic modulus
(EM). The TS of pure FG lm was 7.44 0.17 MPa, which increased
to a maximum value of 11.28 1.02 MPa when the CSNPs content
was 8% (w/w) (Table 1). However, the increase was not signicant
(p > 0.05) below 4% (w/w) CSNPs content. Addition of CSNPs
resulted in a stiffer material conrming the reinforcing effect of the
nanoparticle in the polymeric matrix, which is consistent with
previously reported results (Chang et al., 2010; De Moura et al.,
2009). In these ways, Chang et al. (2010) reported that the addition of CSNPs to glycerol plasticised-starch matrix results in
improvement of TS in the lms. Compared with CSNPs, addition of
chitosan into walleye pollock-skin gelatin was found to have
obvious effect of improving the TS property of resultant materials
(Liu et al., 2011). The TS of packaging lm must be more than
3.5 MPa, according to conventional standards (Kim, Lee, & Park,
1995). Thus, the value of over 11 MPa for FG-CSNPs (8%, w/w)
lm is a super value for its use as a disposable packaging lm. For
elongation at break (EAB), a decreasing trend was found. The EAB
value decreased from 102.04 28.38% to 32.73 7.38% by
increasing in the content of CSNPs. Decrease in EAB is an indication
of an increase in the brittleness of the gelatin lms. A decrease in
elongation of sh gelatin by incorporation of chitosan was also

mez-Estaca et al. (2011). This has been reported to
observed by Go
be also a result of decreases in the free volume between polymer
chains caused by an increase in intermolecular attractive forces,

Fig. 9. Typical stress-strain curves of FG-based nanocomposite lms with different

CSNPs content.

making the polymer network high dense and thus less permeable.
Therefore, direct interaction and proximity between protein chains
gets reduced, which led to the decrease of EAB (Jeya Shakila et al.,
2012). Elastic modulus (EM), a measure of intrinsic lm stiffness,
is the slope of the linear range of the stress-strain plot (Mauer,
Smith, & Labuza, 2000). As can be seen from Table 1, the EM of
the control FG lm was 287.03 14.25 MPa. With increasing content of CSNPs, the EM was found to be elevated from 287.03 14.25
to 467.2 49.63 MPa, which was an improvement by ~60%. Similarly, De Moura et al. (2009) reported that the blending of CS-TPP
nanoparticles with HPMC lms exhibited increased EM of the
lms. Moreover, EM values of the nanocomposite lms in the
present study were higher than those for chitosan incorporating
grouper bone gelatin lms, which have been reported 378 MPa
(Jeya Shakila et al., 2012). Increased elastic modulus can signicantly be serviceable in the food packaging application.
3.7. Water solubility
Solubility is an important property of edible lms as they are
used as protective layers on food. Potential applications may
require water insolubility to enhance product integrity and water
resistance (De Moura, Lorevice, Mattoso, & Zucolotto, 2011). As can
be seen in Table 1, the solubility of pure FG lms in distilled water
was around 72%, that was in accordance with the values reported
by Jiang, Liu, Du, and Wang (2010) for lms elaborated from catsh
skin gelatin and were considerably lower than those reported for
~ ez-Flores et al., 2012). By
cod skin gelatin lms of around 20% (Nn
addition of CSNPs to the lm matrix, the observed decrease in water

Table 1
Tensile strength (TS), elongation at break (EAB), elastic modulus (EM), lm solubility and water vapor permeability (WVP) of FG-based nanocomposite lms with different
CSNPs content.
CSNPs content (%)
0
2
4
6
8

Thickness (mm)
50.58
54.52
61.16
62.14
64.87

2.80
4.10a
2.48b
1.67b
3.36b

TS (MPa)
7.44
7.99
8.77
10.57
11.28

EAB (%)
a

0.17
1.46a
1.11ab
0.19b
1.02bc

102.04
70.09
64.72
44.71
32.73

EM (MPa)
a

28.38
11.93ab
24.59b
11.80b
7.38b

287.03
371.93
392.25
453.46
467.2

Film solubility (%)

a

14.25
4.61b
6.43bc
65.46c
49.63c

71.80
68.55
63.79
62.63
65.19

1.51
2.67a
0.15b
1.14b
2.32ab

WVP (g mm/kPa h m2)

1.421
1.006
0.832
0.717
0.884

0.087a
0.170b
0.038bc
0.036c
0.127bc

Reported values for each lm are means standard deviation. Values means followed by the same letter are not signicantly (p > 0.05) different according LSD test.

180

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

slightly increased. This trend can be explained to this way that at

low levels, CSNPs could disperse well in the FG matrix (as shown by
SEM), thereby blocking the water vapor. However, as illustrated by
SEM micrographs, additional CSNPs (i.e. 8%, w/w) aggregated easily,
which actually decreased the effective content of CSNPs and facilitated water vapor permeation. Similar effects on the water vapor
behavior of glycerol plasticised-starch/CSNPs composites have
been reported by Chang et al. (2010).

solubility can be attributed to the formation of strong hydrogen

bonds between gelatin matrix and the nanoparticles, which
conrmed by FTIR results. Water molecules are not able to break
this bond sufciently hence the solubility of FG-CSNPs lm with 6%
(w/w) CSNPs was the lowest (Table 1). Our results are comparable
to a previous report by De Moura et al. (2011) who have reported a
reduction in the water solubility on addition of chitosan nanoparticles to the methylcellulose lms. Voon, Bhat, Easa, Liong, and
Karim (2012) also found that the addition of the nanoclay in
bovine gelatin lm matrix decreased the water solubility, and they
explained that this could be due to the formation of strong
hydrogen bonds between gelatin matrix and the nanoparticles. In
comparison with CSNPs effect on FG lm in the present study, the
water solubility of tuna-skin gelatin lm was signicantly reduced

mez-Estaca et al., 2011). This
by adding chitosan biopolymer (Go
improvement was attributed to the formation of polyelectrolyte
complexes (PECs) between the ammonium groups of chitosan and
the negatively charged side-chain groups in the gelatin (i.e.
carboxylate groups) through electrostatic interaction (Yin, Li, Sun,
& Yao, 2005).

3.9. Light transmission and transparency

Transparency of lms is greatly relevant to the lms functionality due to their great impact on the appearance of the products

nechal, & Orts, 2011). Usually,
(Bilbao-Sainz, Bras, Williams, Se
higher transparency is considered to be desirable for food packaging lms and coatings, since consumers prefer to see foods. Light
transmission and transparency of control FG lm and FG-based
nanocomposite lms at selected wavelengths are shown in
Table 2. The transmission of UV light is found very low at
200e280 nm for all the lms. This result is in agreement with
previous reports on sh gelatin based lms (Hosseini et al., 2013;
Jiang et al., 2010). According to Aitken and Learmonth (1996),
barrier properties of proteins against UV radiation are associated
with the presence of UV-absorbing chromophore, especially aromatic amino acids-tyrosine and tryptophan and in a less extent,
phenylalanine and disulde bonds. Moreover, the addition of CSNPs
led to the decline of light transmission in the UV region; depend on
the amount of particles added as shown in Table 2. Since CSNPs are
not dissolved in the lm solution, its addition to the lm matrix
causes a decrease in the transparency of the lms. As a consequence
the opaque appearance of the FG-CSNPs composite lms reects
the UV light, thereby hinders light transmission through the lms.
Therefore, FG-CSNPs lms have excellent barrier properties against
UV light, which induces lipid oxidation in the food system
(Coupland & McClements, 1996). When comparing bionanocomposite prepared lms with composite lms based on sh
gelatin and chitosan (Hosseini et al., 2013), the better light barrier
properties of FG lms containing CSNPs than those containing
chitosan have been found. As shown in Table 2, in visible range
(350e800 nm), all bio-nanocomposite lms showed the lower light
transmission than the control. As a result of the increased light
blocking properties (as discussed above), lms with CSNPs also had
lower transparency values than pure gelatin lms (Table 2). The
transparency of the FG-based lms decreased signicantly
(p < 0.05) when the lm was blended with the CSNPs. This decrease
was found to depend on the amount of CSNPs added as shown in
Table 2. The reason that the lms lost transparency when adding
the CSNPs at higher contents (i.e. 8%, w/w) might be due to the
aggregation of nanoparticles which, in turn, obstruct the transmission of light. The effect of CSNPs on reducing the transparency
of FG lms is similar to the effect of cellulose nanoparticles on
transparency of HPMC lms (Bilbao-Sainz et al., 2011).

3.8. Water vapor permeability (WVP)

The problem in composite lms used in food industry is the
relatively high-water vapor permeability of the edible lms.
Permeability in lms is controlled by the diffusivity and solubility of
water molecules within the lm matrix (Gontard et al., 1992). Thus
using nanoscience, new forms of tightly linked three dimensional
networks can be developed to prevent migration of water in food
products (De Moura et al., 2009). The effect of nanoparticle contents on water vapor permeability (WVP) of FG lms is given in
Table 1. The WVP of the control FG lm with no particle inclusions
was 1.421 0.087 g mm/kPa h m2, which is comparable to those for
lms based on sh gelatin as reported in literature (Avena-Bustillos
et al., 2006; Ninan, Joseph, & Abubacker, 2010). The addition of
CSNPs signicantly decreased the WVP of FG lm (Table 1), as this
decline for 6% (w/w) CSNPs has been of about 50% (p < 0.05). According to Chang et al. (2010), the addition of CSNPs produces a
tortuous path that hinders the passage of water molecules through
the lm matrix. Again, the decrease in WVP of lms could be
attributed to the restricted mobility of protein molecules due to the
incorporation of nanoparticles and its interaction with gelatin
(Vanin et al., 2014). On the other hand, De Moura et al. (2009) reported that formation of hydrogen bonding between CS-TPP
nanoparticles and the HPMC lm matrix decreased the WVP of
lms to a considerable extent. In another study, the effectiveness of
the reinforcing effect of chitosan nanoparticles in banana puree
lms was described by Martelli et al. (2013). The results from this
study demonstrated that the incorporation of nanoparticles acted
in reducing the water vapor permeation rate, which has been
attributed to the diffusion of chitosan nanoparticles during solvent
evaporation and lling empty spaces in the lm matrix. When more
than 6% (w/w) CSNPs was added to the FG matrix, WVP values

Table 2
Light transmission (%) and transparency of FG-based nanocomposite lms with different CSNPs content.
CSNPs content (%)

Light transmission at different wavelengths (%)

200

0
2
4
6
8

0
0
0
0
0

280
0.00a
0.00a
0.00a
0.00a
0.00a

14.52
11.30
10.83
9.58
7.76

350

0.36a
0.57b
0.44bc
1.12cd
0.91e

69.94
68.34
66.67
54.05
43.01

Transparency
400

1.37a
1.47a
2.57a
3.82b
0.63c

77.79
75.03
73.7
63.95
49.4

500

1.62a
2.40a
1.48a
4.38b
1.00c

80.54
80.07
78.88
72.48
53.44

600

1.06a
1.07a
2.48a
2.44b
1.12c

82.51
81.56
80.26
73.84
55.34

800

1.02a
1.37a
1.51a
4.15b
1.12c

83.24
82.53
81.43
76.94
57.24

0.95a
1.32a
0.89a
3.57b
1.02c

0.97
0.99
1.29
1.61
3.65

0.03a
0.01a
0.18a
0.23b
0.08c

Reported values for each lm are means standard deviation. Values means followed by the same letter are not signicantly (p > 0.05) different according LSD test.

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

4. Conclusions
In this research, bio-nanocomposite lms based on FG and
CSNPs have been successfully developed. The particles were
spherical in shape with size range 40e80 nm. The incorporation of
CSNPs to FG lms improved their water vapor barrier, as well as TS
and elastic modulus, indicating that the nanoparticles improve the
lm applicability as edible packaging. SEM images revealed that
CSNPs was dispersed evenly in the FG matrix at lower loading
levels. These ndings indicate that use of nanotechnology can
improve functionality to edible lms for food applications. However, relatively high WVP and lm solubility values of these lms as
compared to those of LDPE and PVDC might limit the application of
these bio-nanocomposite lms to packaging of high moisture
foods. Hence, there is a need to further improve the properties of
these nanocomposite lms for commercial application.
Acknowledgment
Authors would like to thank the Iran National Science Foundation (INSF) (Project No. 90000234) and Iran Nanotechnology
Initiative council (INIC) for nancial support of this research.
References
Ahmed, J., Varshney, S. K., & Auras, R. (2010). Rheological and thermal properties of
polylactide/silicate nanocomposites lms. Journal of Food Science, 75, 17e24.
Aitken, A., & Learmonth, M. P. (1996). Protein determination by UV absorption. In
J. M. Walker (Ed.), The protein protocols handbook (pp. 3e6). Totowa, NJ: Human
Press Inc.
Ali, S. W., Rajendran, S., & Joshi, M. (2011). Synthesis and characterization of chitosan and silver loaded chitosan nanoparticles for bioactive polyester. Carbohydrate Polymers, 83, 438e446.
ASTM. (2005). Standard test method for water vapor transmission of materials (E 9605). Philadelphia, PA, USA.
ASTM. (2009). Standard test method for tensile properties of thin plastic sheeting (D
882-09). Philadelphia, PA, USA.
Avena-Bustillos, R. J., Olsen, C. W., Olson, D. A., Chiou, B., Yee, E., Bechtel, P. J., et al.
(2006). Water vapor permeability of mammalian and sh gelatin lms. Journal
of food science, 71, 202e207.
Badii, F., & Howell, N. K. (2006). Fish gelatin: structure, gelling properties and
interaction with egg albumen proteins. Food Hydrocolloids, 20, 630e640.
Bae, H. J., Park, H. J., Hong, S. I., Byun, Y. J., Darby, D. O., Kimmel, R. M., et al. (2009).
Effect of clay content, homogenization RPM, pH, and ultrasonication on mechanical and barrier properties of sh gelatin/montmorillonite nanocomposite
lms. LWT e Food Science and Technology, 42, 1179e1186.
Bergo, P., & Sobral, P. J. A. (2007). Effects of plasticizer on physical properties of
pigskin gelatin lms. Food Hydrocolloids, 21, 1285e1289.

nechal, T., & Orts, W. (2011). HPMC reinBilbao-Sainz, C., Bras, J., Williams, T., Se
forced with different cellulose nano-particles. Carbohydrate Polymers, 86,
1549e1557.
~ na

n-Lo

pez, C., & Vila-Jato Alonso, M. J. (1997). Novel hydrophilic
Calvo, P., Remu
chitosan-polyethylene oxide nanoparticles as protein carrier. Journal of Applied
Polymer Science, 63, 125e132.
Casariego, A., Souza, B. W. S., Cerqueira, M. A., Teixeira, J. A., Cruz, L., Daz, R., et al.
(2009). Chitosan/clay lms' properties as affected by biopolymer and clay micro/nanoparticles' concentrations. Food Hydrocolloids, 23, 1895e1902.
Chang, P. R., Jian, R., Yu, J., & Ma, X. (2010). Fabrication and characterisation of
chitosan nanoparticles/plasticised-starch composites. Food Chemistry, 120,
736e740.
Coupland, J. N., & McClements, D. J. (1996). Lipid oxidation in food emulsions. Trends
in Food Science and Technology, 7, 83e91.
Cuq, B., Gontard, N., & Guilbert, S. (1998). Proteins as agricultural polymers for
packaging production. Cereal Chemistry, 75, 1e9.
De Moura, M. R., Aouada, F. A., Avena-Bustillos, R. J., McHugh, T. H., Krochta, J. M., &
Mattoso, L. H. C. (2009). Improved barrier and mechanical properties of novel
hydroxypropyl methylcellulose edible lms with chitosan/tripolyphosphate
nanoparticles. Journal of Food Engineering, 92, 448e453.
De Moura, M. R., Lorevice, M. V., Mattoso, L. H. C., & Zucolotto, V. (2011). Highly
stable, edible cellulose lms incorporating chitosan nanoparticles. Journal of
Food Science, 76, 25e29.
Derjaguin, B. V., & Landau, L. (1993). Theory of the stability of strongly charged
lyophobic sols and of the adhesion of strongly charged particles in solutions of
electrolytes. Progress in Surface Science, 43, 30e59.
Dudhani, A. R., & Kosaraju, S. L. (2010). Bioadhesive chitosan nanoparticles: preparation and characterization. Carbohydrate Polymers, 81, 243e251.

181

Endres, H. J., & Siebert-Raths, A. (2012). Performance prole of biopolymers

compared to conventional plastics. In M. Moeller, & K. Matyjaszewski (Eds.),
Polymer science: A comprehensive reference (pp. 317e353). Amsterdam, Netherland: Elsevier Science.
Frone, A. N., Berlioz, S., Chailan, J. F., & Panaitescu, D. M. (2013). Morphology and
thermal properties of PLA-cellulose nanobers composites. Carbohydrate Polymers, 91, 377e384.

mez-Estaca, J., Go

mez-Guille

n, M. C., Fern

Go
andez-Martn, F., & Montero, P. (2011).
Effects of gelatin origin, bovine-hide and tuna-skin, on the properties of compound gelatin-chitosan lms. Food Hydrocolloids, 25, 1461e1469.

mez-Guille

n, M. C., Pe

rez-Mateos, M., Go

mez-Estaca, J., Lo

pez-Caballero, E.,
Go

nez, B., & Montero, P. (2009). Fish gelatin: a renewable material for
Gime
developing active biodegradable lms. Trends in Food Science and Technology,
20, 3e16.
Gontard, N., Guilbert, S., & Cuq, J. L. (1992). Edible wheat gluten lms: Inuence of
the main process variables on lm properties using response surface methodology. Journal of Food Science, 57, 190e195.
Han, J. H., & Floros, J. D. (1997). Casting antimicrobial packaging lms and
measuring their physical properties and microbial activity. Journal of Plastic Film
and Sheeting, 13, 287e298.
Hosseini, S. F., Rezaei, M., Zandi, M., & Farahmandghavi, F. (2013). Preparation and
functional properties of sh gelatin-chitosan blend edible lms. Food Chemistry,
136, 1490e1495.
Huq, T., Salmieri, S., Khan, A., Khan, R. A., Tien, C. L., Riedl, B., et al. (2012). Nanocrystalline cellulose (NCC) reinforced alginate based biodegradable nanocomposite lm. Carbohydrate Polymers, 90, 1757e1763.
Jeya Shakila, R., Jeevithan, E., Varatharajakumar, A., Jeyasekaran, G., & Sukumar, D.
(2012). Comparison of the properties of multi-composite sh gelatin lms with
that of mammalian gelatin lms. Food Chemistry, 135, 2260e2267.
Jiang, M., Liu, A., Du, X., & Wang, Y. (2010). Physical properties and internal
microstructure of lms made from catsh skin gelatin and triacetin mixtures.
Food Hydrocolloids, 24, 105e110.
Jingou, J., Shilei, H., Weiqi, L., Danjun, W., Tengfei, W., & Yi, X. (2011). Preparation,
characterization of hydrophilic and hydrophobic drug in combine loaded chitosan/cyclodextrin nanoparticles and in vitro release study. Colloids and Surfaces
B: Biointerfaces, 83, 103e107.
Kanmani, P., & Rhim, J. W. (2014). Physical, mechanical and antimicrobial properties
of gelatin based active nanocomposite lms containing AgNPs and nanoclay.
Food Hydrocolloids, 35, 644e652.
Khan, A., Khan, R. A., Salmieri, S., Tien, C. L., Riedl, B., Bouchard, J., et al. (2012).
Mechanical and barrier properties of nanocrystalline cellulose reinforced chitosan based nanocomposite lms. Carbohydrate Polymers, 90, 1601e1608.
Kim, Y. J., Lee, H. M., & Park, O. O. (1995). Processabilities and mechanical properties
of Surlyn-treated starch/LDPE blends. Polymer Engineering and Science, 35,
1652e1657.
Kumar, P., Sandeep, K. P., Alavi, S., Truong, V. D., & Gorga, R. E. (2010). Effect of type
and content of modied montmorillonite on the structure and properties of
bio-nanocomposite lms based on soy protein isolate and montmorillonite.
Journal of Food Science, 75, 46e56.
Le Tien, C., Letendre, M., Ispas-Szabo, P., Mateescu, M. A., Delmas-Patterson, G.,
Yu, H. L., et al. (2000). Development of biodegradable lms from whey proteins
by cross-linking and entrapment in cellulose. Journal of Agricultural Food and
Chemistry, 48, 5566e5575.
Liu, Y., Sun, Y., Li, Y., Xu, S., Tang, J., Ding, J., et al. (2011). Preparation and characterization of a-galactosidase-loaded chitosan nanoparticles for use in foods.
Carbohydrate Polymers, 83, 1162e1168.
Lorevicel, M. V., De Moura, M. R., Aouada, F. A., & Mattoso, L. H. C. (2012). Development of novel guava puree lms containing chitosan nanoparticles. Journal of
Nanoscience and Nanotechnology, 12, 2711e2717.
~ a, L. N., Alvarez, V. A., & Vasquez, A. (2007). Processing and microstructure of
Luduen
PCL/clay nanocomposites. Materials Science and Engineering A, 460-461, 121e129.
Ma, X. F., Yu, J. G., & Wang, N. (2008). Glycerol plasticized-starch/multiwall carbon
nanotube composites for electroactive polymers. Composites Science and Technology, 68, 268e273.
Martelli, M. R., Barros, T. T., De Moura, M. R., Mattoso, L. H. C., & Assis, O. B. G. (2013).
Effect of chitosan nanoparticles and pectin content on mechanical properties
and water vapor permeability of banana puree lms. Journal of Food Science, 78,
98e104.
Mauer, L. J., Smith, D. E., & Labuza, T. P. (2000). Water vapor permeability and
mechanical properties of edible b-casein lms. International Dairy Journal, 10,
353e358.
McHugh, T. H., Avena-Bustillos, R., & Krochta, J. (1993). Hydrophilic edible lms:
modied procedure for water vapour permeability and explanation of thickness
effects. Journal of Food Science, 58, 899e903.
McHugh, T. H., & Krochta, J. M. (1994). Permeability properties of edible lms. In
J. M. Krochta, E. A. Baldwin, & M. O. Nisperos-Carriedo (Eds.), Edible coatings and
lms to improve food quality (pp. 139e187). Lancaster, PA, USA: Technomic
Publishing.
Murillo-Martnez, M. M., Pedroza-Islas, R., Lobato-Calleros, C., Martnez-Ferez, A., &
Vernon-Carter, E. J. (2011). Designing W1/O/W2 double emulsions stabilized by
protein-polysaccharide complexes for producing edible lms: Rheological,
mechanical and water vapour properties. Food Hydrocolloids, 25, 577e585.
Ninan, G., Joseph, J., & Abubacker, Z. (2010). Physical, mechanical, and barrier
properties of carp and mammalian skin gelatin lms. Journal of Food Science, 75,
620e626.

182

S.F. Hosseini et al. / Food Hydrocolloids 44 (2015) 172e182

~ ez-Flores, R., Gime

nez, B., Ferna

ndez-Martn, F., Lo

pez-Caballero, M. E.,
Nn

mez-Guille

n, M. C. (2012). Role of lignosulphonate in
Montero, M. P., & Go
properties of sh gelatin lms. Food Hydrocolloids, 27, 60e71.
Nur Hanani, Z. A., Roos, Y. H., & Kerry, J. P. (2012). Use of beef, pork and sh gelatin
sources in the manufacture of lms and assessment of their composition and
mechanical properties. Food Hydrocolloids, 29, 144e151.
Pereda, M., Ponce, A. G., Marcovich, N. E., Ruseckaite, R. A., & Martucci, J. F. (2011).
Chitosan-gelatin composites and bi-layer lms with potential antimicrobial
activity. Food Hydrocolloids, 25, 1372e1381.
Qi, L., Xu, Z., Jiang, X., Hu, C., & Zou, X. (2004). Preparation and antibacterial activity
of chitosan nanoparticles. Carbohydrate Research, 339, 2693e2700.
Rokhade, A. P., Agnihotri, S. A., Patil, S. A., Mallikarjuna, N. N., Kulkarni, P. V., &
Aminabhavi, T. M. (2006). Semi-interpenetrating polymer network microspheres of gelatin and sodium carboxymethyl cellulose for controlled release of
ketorolac tromethamine. Carbohydrate Polymers, 65, 243e252.
Shahidi, F., Arachchi, J. K. V., & Jeon, Y. J. (1999). Food applications of chitin and
chitosan. Trends in Food Science and Technology, 10, 37e51.
Sionkowska, A., Wisniewski, M., Skopinska, J., Kennedy, C. J., & Wess, T. J. (2004).
Molecular interactions in collagen and chitosan blends. Biomaterials, 25,
795e801.

Vanin, F. M., Hirano, M. H., Carvalho, R. A., Moraes, I. C. F., Bittante, A. M. Q. B., &
Sobral, P. J. A. (2014). Development of active gelatin-based nanocomposite lms
produced in an automatic spreader. Food Research International, 63, 16e24.
Verwey, E. J. W., & Overbeek, J. Th G. (1948). Theory of the stability of lyophobic
colloids. Amsterdam: Elsevier.
Voon, H. C., Bhat, R., Easa, A. M., Liong, M. T., & Karim, A. A. (2012). Effect of addition
of halloysite nanoclay and SIO2 nanoparticles on barrier and mechanical
properties of bovine gelatin lms. Food and Bioprocess Technology, 5, 1766e1774.
Yang, H. C., Wang, W. H., Huang, K. S., & Hon, M. H. (2010). Preparation and
application of nanochitosan to nishing treatment with anti-microbial and
anti-shrinking properties. Carbohydrate Polymers, 79, 176e179.
Yin, Y., Li, Z., Sun, Y., & Yao, K. (2005). A preliminary study on chitosan/gelatin
polyelectrolyte complex formation. Journal of Material Science, 40, 4649e4652.
Yoksan, R., Jirawutthiwongchai, J., & Arpo, K. (2010). Encapsulation of ascorbyl
palmitate in chitosan nanoparticles by oil-in-water emulsion and ionic gelation
processes. Colloids and Surfaces B: Biointerfaces, 76, 292e297.
Zheng, J. P., Li, P., Ma, Y. L., & Yao, K. D. (2002). Gelatin/montmorillonite hybrid
nanocomposite I. Preparation and properties. Journal of Applied Polymer Science,
86, 1189e1194.