International Journal of Biological Macromolecules 185 (2021) 1005–1014

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International Journal of Biological Macromolecules

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

Effect of lipids with different physical state on the physicochemical

properties of starch/gelatin edible films prepared by extrusion blowing
Yue Cheng a, 1, Cong Sun a, 1, Xiaosong Zhai a, Rui Zhang a, Shikai Zhang a, Chanchan Sun b,
Wentao Wang a, *, Hanxue Hou a, *
a
Department of Food Science and Engineering, Shandong Agricultural University, Tai'an 271018, China
b
Key Laboratory of Food Nutrition and Safety (Tianjin University of Science &Technology), Ministry of Education, Tianjin 300457, China

A R T I C L E I N F O A B S T R A C T

Keywords: The effects of various physical state lipids (rapeseed oil (RO), shortening (ST), beeswax (BW)), on the physi­
Starch cochemical properties of starch (S) (hydroxypropyl distarch phosphate (HP), oxidized hydroxypropyl starch
Gelatin (OS))/gelatin (G) blown films were studied. S/G-lipid blends showed decreased storage modulus and complex
Lipids
viscosity. The formation of hydrogen bonds was inhibited by the ST and BW, but facilitated by the RO. Compared
Edible film
with BW and ST, RO was more effective to promote the melted and fractured of starch. Lipids addition promoted
Extrusion blowing
the compatibility of starch and gelatin. The presence of the lipids significantly improved the surface hydro­
phobicity, mechanical, water vapor barrier and water resistance properties of S/G films. S/G-RO films exhibited
the strongest surface hydrophobicity and tensile strength, while HP/G-BW film showed the strongest water
resistance and water vapor barrier properties. These results revealed that the appropriate lipids could be used to
produce S/G-lipid films with desirable physicochemical properties.

1. Introduction
Edible film is a thin layer used for primary food packaging and can be
consumed with packaged food. The edible film can provide a protective
layer on the food surface to control the selective exchange of moisture,
flavor and important gases, as well as solute movement out of the food
[1]. Compared with the currently widely used petroleum-based films,
edible films based on natural biopolymers have received widespread
attention in food packaging and storage because of safe edibility and
biodegradability [2]. To date, the film-forming materials used for edible
films have mainly included polysaccharides, proteins and lipids. Among
these materials, starch, as a polysaccharide with good film-forming
properties, has promising application prospects in the field of edible
packaging due to its low cost, easy availability and rapid biodegrad­
ability [3]. However, the application of starch films has been limited
because of their strong hydrophilicity which usually results in poor
water vapor barrier properties and water resistance.
The addition of lipids into edible films is generally considered to
reduce their hydrophilicity and make them more suitable for food
packaging [4]. Lipids have been shown to form a hierarchical structure
of solidified wax on the film surface when beeswax was incorporated
into starch-based films, which significantly improved their hydrophobic
properties [5]. However, the mechanical and water vapor barrier
properties of starch film were weakened due to the poor compatibility of
between starch and lipid. Gelatin could theoretically be used to combine
starch with lipids to form films because of hydrophilic-lipophilic
(amphipathic) character of gelatin. We have previously reported that
the addition of gelatin into starch-lipid films significantly improved
their mechanical and water vapor barrier properties, and found that the
physicochemical properties of these films were affected by the particle
size and distribution of lipids [6]. Therefore, it is necessary to further
explore the effects of the lipids on the structure and properties of starch/
gelatin (S/G) films in obtaining the desired properties of the end prod­
ucts used in practical applications. At present, the lipids used for edible
films mainly include oils, fats and waxes, which generally have different
physical state (liquid, semi-solid and solid) [7]. These three types of
lipids possibly had a remarkable influence on the size and distribution of
lipids in film matrix due to their different melting points [8]. Oil exists in
melted state and easily exudates on film surface during the film molding
process, while pre-melted fat and wax show a quick crystallization

* Corresponding authors at: Department of Food Science and Engineering, Shandong Agricultural University, Tai'an City, Shandong Province 271018, China.
E-mail addresses: wangwt@sdau.edu.cn (W. Wang), hhx@sdau.edu.cn (H. Hou).
1
Yue Cheng and Cong Sun contributed equally to this work and are co-first authors.

https://doi.org/10.1016/j.ijbiomac.2021.06.203
Received 14 March 2021; Received in revised form 4 June 2021; Accepted 29 June 2021
Available online 2 July 2021
0141-8130/© 2021 Published by Elsevier B.V.
Y. Cheng et al.
which disturbs film structure, leading to a negative impact on the per­
formance of films. Zhang et al. [9] reported that three hydrophobic
agents with different physical state had different effects on the physical
and mechanical properties of agar/maltodextrin films. However, the
effects of lipids with different physical state on the properties of S/G
films, to the best of our knowledge, have been hardly investigated.
To date, edible films have been mainly prepared by solution casting,
which is only adequate for laboratory research and cannot be
commercialized [10]. Moreover, the addition of solid lipids usually
makes the solution casting process difficult because high temperature is
required for the melting. Extrusion blowing is an interesting process in
offering high temperatures required during film-forming process, which
can increase the commercial potential of edible films. In this study,
various lipids (rapeseed oil (RO), shortening (ST) and beeswax (BW))
with different physical states were incorporated in starch/gelatin films
to obtain a highly hydrophobic edible film with higher mechanical and
water vapor barrier properties. The effects of three lipids on the struc­
tures, physicochemical properties, and film-forming mechanism of S
(hydroxypropyl distarch phosphate (HP), oxidized hydroxypropyl starch
(OS))/G films were assessed.
2. Materials and methods
2.1. Materials
Cassava starch (HP and OS) samples were provided by Puluoxing
Starch Co., Ltd. (Hangzhou, China). The HP contained 12.5 wt% mois­
ture, 3.1 wt% hydroxypropyl group, 0.0 wt% phosphate content, and
22.5 wt% amylose. The OS contained 11.8 wt% moisture, 2.0 wt%
hydroxypropyl group, 0.3 wt% carboxyl groups, 0.1 wt% carbonyl
content, and 22.9 wt% amylose. Bovine Gelatin (180 Bloom value) was
obtained from Shangshui county Fu'yuan Gelatin Co., Ltd. (Zhoukou,
China). RO (Melting point: − 10 to − 6 ◦ C), ST (Melting point: 48–53 ◦ C)
and BW (Melting point: 62–65 ◦ C) was provided by Yihai Grain and Oil
Industry Co., Ltd. (Yantai, China). Glycerol was purchased from Tianjin
Fuyu Chemical Co., Ltd. (Tianjin, China). Tween-80 was bought from
Lemon Chemical & Technology Co., Ltd. (Wuxi, China).
2.2. Preparation of the films
HP and OS (2000 g) were mixed with gelatin (1000 g) for 5 min using
an SHR50A mixer (Hongji Machinery, Zhangjiagang, China), respec­
tively. RO (180 g), ST (180 g), and BW (180 g) were mixed with glycerol
Fig. 1. Continuous and stable film blowing of S/G-lipid films.
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International Journal of Biological Macromolecules 185 (2021) 1005–1014
(900 g), deionized water (250 g) and Tween 80 (90 g), respectively, and
homogenized for 5 min at 20,000 rpm using an IKA Ultra Turrax T18
digital homogenizer (IKA, Germany). The solution obtained was incor­
porated into the S/G mixture prepared and mixed for 5 min. The six
samples obtained were coded by the types of starch and lipid, such as
HP/G-RO, HP/G-ST, and HP/G-BW, OS/G-RO, OS/G-ST and OS/G-BW.
The control sample was prepared without the addition of natural waxes
with the above-mentioned procedure.
Resulting mixtures were compounded using an SHJ-20B laboratory
twin-screw extruder (Giant Machinery, Nanjing, China) with a screw
diameter (D) of 21.7 mm and length of 40 D. The screw speed was 130
rpm, and the barrel temperatures from the feed zone to the die zone were
set to 95, 105, 110, 120, 105, and 95 ◦ C. The extrudates was air-cooled
and cut into pellets.
S/G films were prepared using an SCM-50 laboratory film-blowing
extruder (Lianjiang Machinery, Zhangjiagang, China) with a D of 25
mm, length of 30 D, and die diameter of 30 mm. The screw speed was 25
rpm, and the extrusion temperatures were set to 80, 110, 120, 125, 115,
and 95 ◦ C. The film thickness was controlled at 85–95 μm by carefully
altering the ratios of blow-up and take-up. All measurements were
completed in 8–10 days after the preparation of samples. Continuous
and stable film blowing of S/G composite films was shown in Fig. 1.
2.3. Rheological measurements of S/G blends
The pellets made in Section 2.2 were hot-pressed into film samples
with thickness of 1 mm for rheological analysis. The rheological prop­
erties of all S/G samples were examined at a fixed temperature of 120 ◦ C
using a MCR 102 modular intelligent rheometer (Anton Paar, Austria)
with a 50-mm parallel plate. To achieve thermal equilibrium and
structural relaxation, a waiting time was applied after the loading and
prior to measurements. Dynamic strain sweeps revealed that the results
of rheological measurements performed at a strain of 1% safely fell
within the linear viscoelastic range. The complex viscosity (|η*|) and
storage modulus (G′ ) were determined in a frequency range of 0.01–100
rad/s. The water evaporation was suppressed with mineral oil in the
process of testing.
2.4. Attenuated total reflectance–Fourier transform infrared (ATR–FTIR)
spectroscopy
ATR-FTIR spectroscopy spectra (Nicolet IS 10, Thermo Electron,
USA) of film samples was used for characterization. The absorption
Y. Cheng et al.
spectra were acquired over a wavenumber range of 4000–400 cm− 1 with
32 scans and a resolution of 4.0 cm− 1 a resolution against the back­
ground. Finally, the obtained curves were normalized.
2.5. X-ray diffraction (XRD)
The XRD patterns of films were examined using a Bruker D8 Advance
XRD diffractometer (D8 Advance, Bruker AXS, Germany). The test
samples were analyzed at 25 ◦ C with a diffraction angle (2θ) between 5
and 40◦ at a step of 0.02◦ /s. Relative crystallinity of the samples was
evaluated as follows:
Ac
Relative crystallinity (%) = × 100
Aa + Ac
where Aa and Ac are the amorphous and crystalline areas, respectively.
2.6. Scanning electron microscopy (SEM)
The morphological features of the surface and cross-section of the
films were examined using scanning electron microscopy (SEM, Supra
55, Zeiss, Germany). To observe the cross-sections, the films were
fractured immediately after being treated with liquid nitrogen. The
images were acquired at an acceleration voltage of 3 kV and 1000×
magnification.
2.7. Dynamic mechanical analysis (DMA)
Dynamic mechanical properties of S/G films were conducted on a
Q800 dynamic mechanical analyzer (TA Instruments, New Jersey, USA).
The films were cut into rectangular shapes with dimensions of 60 mm ×
8 mm. Testing was carried out in the tensile mode from 25 to 120 ◦ C at a
heating rate of 3 ◦ C/min, and the frequency was set to 1 Hz. The storage
modulus (E′ ) and loss factor (tan δ) of the samples were measured as a
function of the temperatures. The tested films were equilibrated at 53 ±
2% RH and 23 ± 2 ◦ C for at least 72 h before testing.
2.8. Dynamic contact angle (DCA) measurements
The DCAs of the film samples were measured at the triple solid­
–liquid–air interface by the fixed drop method using a JC-2000C1 con­
tact angle goniometer (OCA15EC, Dataphysics Co., Ltd., Germany). The
surface hydrophilicity and hydrophobicity of the films were expressed
according to the sessile drop method, and the Laplace–Young equation
was used to fit the contour data of the droplets. Finally, the change in the
contact angle within 150 s was also determined with appropriate
software.
2.9. Water-vapor permeability (WVP)
The water vapor permeability (WVP) of the films was measured
using a PERMETM W3/030 automatic WVP tester according to the
method of Cheng et al. (2020) with some slight modifications. The
measurement of WVP was carried out at 90% RH, with a preheating time
and a weighing interval of 4 h and 2 h, respectively.
2.10. Water solubility (WS) and swelling degree (SD)
The film samples were cut into squares (3 cm × 3 cm), and then dried
at 100 ± 5 ◦ C for 12 h to determine the initial dry weights (W1). The
dried samples were immersed in 60 mL of distilled water at room tem­
perature for 24 h. Undissolved portions of films were filtered out, and
the films were weighed after removal of the surface water with blotting
paper (W2). Then the samples were taken out and finally dried at 100 ±
5 ◦ C for another 12 h to get the weight (W3). The SD (%) and WS (%)
values were determined as follows:
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International Journal of Biological Macromolecules 185 (2021) 1005–1014
W2 − W1
SD (%) = × 100
W1
W1 − W3
WS (%) = × 100
W1
2.11. Mechanical properties
The tensile strength (TS, MPa), elongation at break (EAB, %) and
Young's modulus (YM, MPa) of the films were examined using a TA-XT2i
texture analyzer (Stable Micro System Company, UK) based on a pre­
vious method [6]. The films were cut into 60 mm × 15 mm rectangles.
The initial clamping distance and the velocity were 60 mm and 1 mm/s,
respectively. The tested films were equilibrated at 53 ± 2% RH and 23 ±
2 ◦ C for at least 72 h before testing.
2.12. Statistical analysis
The data in the tables and figures are the mean values and standard
deviations of at least six measurements. Statistical analysis of different
samples was analyzed by analysis of variance using SPSS 20. Compari­
sons among the mean values were determined using the Duncan's-mul­
tiple range test at a 5% significance level.
3. Results and discussion
3.1. Rheological properties
Rheological behavior played an important role in polymer process­
ing, especially in extrusion blowing, which required relatively appro­
priate melt strength and fluidity [11]. Changes of complex viscosity (|η
*|) and storage modulus (G′ ) were plotted as functions of angular fre­
quency (�) in Fig. 2. Each material exhibited a typical pseudoplastic
behavior of molten polymers, as occurred a reduction of |η*| values with
the increase of shear frequency (Fig. 2a and b). Such flow behavior is
also called shear-thinning, which is related to the increase in the
orientation degree of polymer molecules and the impairment in chain
entanglement of both starch and gelatin [12]. The reason was that the
higher mechanical stress at higher shear rates during the test led to more
intensive degradation of the polymer molecules, which was manifested
as lower |η*| values of the blends [13], similar to the polymer extrusion
process. All blends with lipids exhibited lower |η*| values than the
control (without lipids), which could improve the fluidity of melts in the
extrusion process [14]. The above-mentioned phenomena could be
caused by the lubrication effect of the lipids that promoted the flow of S/
G blends.
From the perspective of the entire angular frequency range, the G′ of
all samples increased with the increase of frequency, and the de­
pendences were relatively strong (Fig. 2c and d). The G′ values of all
blends with lipids are lower than those of the control at all angular
frequencies, indicating that the addition of lipids decreased the melt
strength of the blends. The lubrication effect of lipids promoted the melt
and plasticization of starch, which facilitated the movement of starch
molecules. Moreover, the increased polymer molecules mobility during
processing promotes phase separation between starch and gelatin,
caused by the intrinsic thermodynamic incompatibility of the two
polymers, which had a negative effect on the G′ of S/G blends [15].
Among all the S/G blends, the |η*| and G′ values of the S/G-RO blends
are obviously lower than those of the blends with others. Based on the
above-mentioned analysis, it is preliminarily speculated that RO
exhibited the best fluidity and lubrication, which was conducive to the
plasticization of starch in the film matrix. In addition, compared with
HP/G blends, OS/G blends exhibited lower |η*| and G′ values which
signified the lower fluidity and less melt strength. This was beneficial for
the processing of blends under certain conditions.
Y. Cheng et al. International Journal of Biological Macromolecules 185 (2021) 1005–1014

Fig. 2. Rheological behaviors of S/G-lipid blends: complex viscosity (|η*|) (a: OS/G-lipid blends, b: HP/G-lipid blends) and storage modulus (G′ ) (c: OS/G-lipid
blends, d: HP/G-lipid blends).

3.2. ATR-FTIR spectra curves presented similar characteristics. As shown in Table 1,

The ATR-FTIR spectra of the composite films with different lipids
were shown in Fig. 3. The broad and intense signal bands around 3288
cm− 1 were ascribed to the O–H stretching vibration [16]. The signifi­
cant bands at 2940 and 2848 cm− 1 came from the C–H stretching vi­
bration [17]. The signal at 1020 cm− 1 is related to C–O stretching
vibrations of C–O–C in starch.
There were some position changes, although all the ATR-FTIR
the bands significantly shifted to higher or lower wavebands after the
addition of lipids. The O–H stretching vibration band shifted to a higher
wavenumber in the presence of BW, indicating that BW disturbed the
film networks via reducing the hydrogen bonds. The intensity of C–H
stretching vibrations band was enhanced because of the addition of BW,
resulting from the fact that BW contains some alkane groups. The similar
phenomenon was discovered in S/G-ST films. However, the O–H
stretching vibration band shifted to lower wavenumber with the

Fig. 3. ATR–FTIR spectra of OS/G-lipid (a) and HP/G-lipid (b) edible films.

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Table 1 plasticization and destroying the crystal structure of starch.

ATR-FTIR data of S/G-lipid films. The XRD peaks for S/G-BW films of 2θ = 21.4◦ –23.6◦ were resulted
Samples T1 T2 from the orthorhombic structure of the hydrocarbon/monoester frac­
c tions [8]. The OS/G-BW film showed weaker peaks at 2θ = 21.4◦ and
OS/G 3285.33 ± 0.58 1019.67 ± 0.58d
OS/G-RO 3282.00 ± 1.00d 1021.33 ± 0.58c 23.6◦ than HP/G-BW film, indicating a reduction in crystalline BW re­
OS/G-ST 3287.00 ± 0.00b 1023.67 ± 0.58b gion. An obvious diffraction peak at 2θ = 6.2◦ for S/G-ST films was
OS/G-BW 3289.67 ± 0.58a 1025.00 ± 1.00a noticed, which was related with ST crystals [20]. No new diffraction
HP/G 3288.00 ± 0.00C 1020.33 ± 0.58B peaks appeared for S/G-RO films because the RO was in a liquid state,
HP/G-RO 3285.33 ± 0.58D 1021.67 ± 0.58A
HP/G-ST 3289.67 ± 0.58B 1021.00 ± 0.00A
which made it hard to form RO crystals.
HP/G-BW 3291.00 ± 0.00A 1021.33 ± 0.58A Crystallinity of the S/G films was also showed in Table 2. The
addition of BW increased the crystallinity of S/G-BW films. The in­
T1 and T2 is the wavenumber of signal bands around 3288 cm− 1 and 1020 cm− 1,
compatibility between BW and polymers during the cooling process led
respectively, corresponding to the peak position in the ATR–FTIR spectra.
a–d
Different lowercase letters means significant difference in comparisons be­
to the formation of BW crystals. The appearance of BW characteristic
tween the lipids (BW, ST, RO) for OS/G-lipid film samples at p < 0.05. diffraction peaks in the S/G-BW films also proved the formation of BW
A–D
Different uppercase letters means significant difference in comparisons be­ crystals (Fig. 4). The crystallinity of S/G-ST films showed the same
tween the lipids (BW, ST, RO) for HP/G-lipid film samples at p < 0.05. trends, although the ST existed in the film matrix as a semi-solid state,
while BW existed as a solid state. This was due to the formation of ST
addition of RO, which indicated that the new hydrogen bonds were crystals. The crystallinity of the S/G-BW films is higher than that of the
formed in the film matrix. This result was attributed to the lubrication S/G-ST films, which may be attributed to the difference in lipid melting
effect of RO, which might facilitate molecular movement and rear­ point. When the cooling temperature of the films was below the melting
rangement of polymers, and thereby formed new hydrogen bonds [17]. point of natural waxes during film formation, the lipids tended to
A molecular interaction took place in edible films when the charac­ crystallize rapidly and randomly, resulting in a random distribution
teristic band shifted after the incorporation of component [8]. As lipids
were added, the C–O stretching vibrations peaks in S/G films shifted
toward a higher wavenumber, indicating that the lipids were able to Table 2
reduce their molecular interaction among the starch granules [18]. Crystallinity, WVP, WS and SD of S/G-lipid films.
These peaks displacement indicated that lipids can cause conformational Samples Crystallinity WVP (10− 13 WS (%) SD (%)
changes and affect the interactions of S/G matrix. By comparison, the (%) g⋅m− 1⋅s− 1⋅Pa− 1)
wavenumber of OS/G-lipid films showed larger shift than HP/G-lipids OS/G 9.12 ± 0.31c 24.26 ± 0.71a 28.82 ± 465.41 ±
films, which indicated that lipids gave rise to stronger conformational 0.37a 9.24a
changes in OS/G films. OS/G- 10.25 ± 0.22b 12.73 ± 0.25b 26.00 ± 324.21 ±
RO 0.35b 5.21b
OS/G-ST 10.58 ± 0.21b 13.80 ± 0.46b 23.12 ± 280.67 ±
3.3. XRD 0.49c 10.28c
a c
OS/G- 11.32 ± 0.13 6.67 ± 0.57 21.69 ± 207.19 ±
The XRD patterns of the S/G films with different lipids were shown in BW 0.62d 8.74d
D A
HP/G 7.14 ± 0.12 25.43 ± 0.81 23.86 ± 400.23
Fig. 4. The characteristic diffraction peaks of gelatin were located at 2θ
±
0.47A 2.27A
= 17◦ –25◦ , derived from the α-helix and triple-dimensional structure of HP/G- 8.06 ± 0.28C 12.43 ± 0.31B 20.56 ± 312.11 ±
gelatin [19]. Starch showed a typical A-type crystalline structure with RO 0.72B 4.23B
main characteristic peaks at 2θ = 15◦ , 17◦ , 18◦ , and 23◦ [6], which was HP/G-ST 8.67. ± 0.19B 13.67 ± 0.71B 20.05 ± 230.24 ±
0.28B 7.56C
partially overlapped with the diffraction peaks of gelatin. Addition of A C
HP/G- 9.31 ± 0.25 4.78 ± 0.52 19.85 ± 187.12 ±
BW and ST caused a decrease in the intensity of the overlapping peak, BW 0.57B 3.12D
but an increase in that of the overlapping peaks in S/G-RO films. These a–d
results indicated that the S/G crystallization was affected by the addi­ Different lowercase letters means significant difference in comparisons be­
tween the lipids (BW, ST, RO) for OS/G-lipid film samples at p < 0.05.
tion of lipids, which was possibly related to the ability to form hydrogen A–D
Different uppercase letters means significant difference in comparisons be­
bonds. Moreover, the addition of lipids reduced the intensity of the
tween the lipids (BW, ST, RO) for HP/G-lipid film samples at p < 0.05.
diffraction peak at 15◦ , which may be due to lipids promoting the

Fig. 4. XRD patterns of OS/G-lipid (a) and HP/G-lipid (b) edible films.

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inside the film. The lipids with higher melting points could reach their dispersibility of lipids in film matrix. It was remarkable that cracks and
crystallization temperature faster, more lipid droplets aggregate, small voids could be seen in the film micrographs, especially for S/G-RO
thereby increasing their crystallinity in S/G films containing lipids with films, which was ascribed to the evaporation of water vapor during the
higher melting points. Because RO promoted the formation of hydrogen film-filming process [22]. Cooling above the lipids melting point could
bonds in the film matrix, the crystallinity of S/G-RO films was also alter the distribution of the lipids after film formation, influencing the
increased compared to that of control film. density of film structure. Hence, lipids were in liquid state which might
tend to flow and spread into film-forming networks as water vapor
evaporated, causing more voids and cracks in S/G-lipid film matrix.
3.4. Morphology
As expected, the presence of unplasticized starch granules resulted in
the rough surface of S/G films with “steamed bread”-like bulges. The
The distribution of lipids in edible film matrix and the interfacial
addition of lipids resulted in greater roughness on the film surface,
interaction with the substrate were directly linked to the performance of
manifesting as many small particles different from the films without
composite films. Fig. 5 showed the microstructures of the S/G composite
lipids. Lipid droplets could be distributed in the film matrix, and partial
films. Many “round zones” appeared in the cross section of the S/G films,
lipid droplets migrated to the surface after the aggregation and floccu­
and their size was approximately 10 μm, which was similar with that of
lation phenomena [23]. Muscat et al. [17] also reported comparable
unplasticized starch granules [21]. According to our report, the starch
result, that is, the small particles of lipid crystal on the film surface
was not completely plasticized in S/G blown films [6]. Thus, the round
significantly increased the surface roughness of the films. Lipids were
zones were caused by the inadequate plasticization of starch granules.
easy to crystallize rapidly and irregularly when the cooling temperature
The microstructure of the films was significantly affected by the addition
of the film was lower than the melting point of lipids, resulting in
of lipids, that is, less unplasticized starch granules were observed in the
irregular distribution in the film matrix [24]. Due to the low melting
S/G-lipid films, indicating the better plasticization of starch. This was
point, RO existed in the film matrix as a liquid with better fluidity during
attributed to the promoting effect of lipids on the melting of film matrix
processing, which promoted the migration of RO to the film surface.
during the processing. The plasticization of starch was beneficial to
Thus, the surface of OS/G-RO film showed more lipid particles than that
interfacial adhesion between starch and gelatin molecules, leading to
of OS/G-BW film. Zhang et al. [8] also reported the similar phenomenon
the formation of a compact film structure. Among all H/G films, H/G-RO
that the surface of films with liquid lipid exhibits higher roughness than
films exhibited the least unplasticized starch granules, which demon­
the films with solid lipid. However, this phenomenon was not found in
strated the speculation of rheological part that RO showed better
HP/G films. This was possibly attributed to that RO aggregated together
lubrication effect on starch than other lipids in the S/G film matrix. No
on the surface of the film to form a layer structure instead of lipid
lipid particles were observed in the S/G-lipid films, exhibiting good

Surface (500h) Cross section (1000h)

(a)

(A) (a) (A) (a)

(B) (b) (B) (b)

(C) (c) (C) (c)

(D) (d) (D) (d)

Fig. 5. SEM images of S/G-lipid films. (A) OS/G, (B) OS/G-RO, (C) OS/G-ST, (D) OS/G-BW, (a) HP/G, (b) HP/G-RO, (c) HP/G-ST and (d) HP/G-BW.

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Y. Cheng et al.
particles.
3.5. DMA
The dynamic mechanical properties of polymers are sensitive to their
molecular motions, which are related to the polymer-chain structure and
molecular interactions [25]. The storage modulus (E′ ) versus tempera­
ture of the S/G films are shown in Fig. 6(a). Strong hydrogen bonding
among polymer molecule chains existed in the control film, which
resulted in the difficulty of movement of the molecular chains, demon­
strating the highest E′ value. With the addition of lipids, the lubrication
effect of the lipid molecules facilitated the movement of the molecular
chains. Consequently, E′ of the films decreased with the addition of RO,
ST and BW, which indicated that lipids addition decreased the rigidity of
the films. As discussed in ATR-FTIR results, RO promoted the formation
of new hydrogen bonds in the film matrix, which hindered the move­
ment of polymer molecular chains. Therefore, E′ values of S/G-RO films
were higher than those of S/G-ST and S/G-BW films.
The variations in the loss factor (tan δ) of the S/G films are shown in
Fig. 6(b). The peak temperature in the tan δ-temperature curve is usually
regarded as the glass-transition temperature (Tg) of the polymer mate­
rial [26]. The shift in their relative positions provided information about
the compatibility of the components. The tan δ-temperature curve of the
S/G films mainly exhibited two peaks, which were associated with the
glass transition of gelatin phase and starch-phase, respectively [6]. This
indicated that the incompatibility of starch and gelatin in the film sys­
tem, which led to two phases: a starch-rich one and a gelatin-rich one.
Compared with the control film, lipid addition led to a decrease in Tg of
starch and an increase in Tg of gelatin, indicating better compatibility of
the starch and gelatin phases. These trends may be attributed from the
Fig. 6. DMA thermograms of the S/G-lipid edible films: storage modulus (E′ ) (a: OS/G-lipid films, c: HP/G-lipid films) and loss factor (tan δ) (b: OS/G-lipid films, d:
HP/G-lipid films).
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International Journal of Biological Macromolecules 185 (2021) 1005–1014
plasticization reaction and interactions between starch and gelatin
molecules [27]. Lipids addition promoted starch plasticization (as
shown in SEM results), which disrupted a larger number of intermo­
lecular hydrogen bonds among the starch granules and decreased the Tg
of starch. In addition, the starch of S/G-lipid films exhibited stronger
hydrophobic properties resulting from the more hydrogen bond expo­
sure, which increased the consumption of plasticizers in the starch
phase. Thus, some glycerol or water may migrate from the gelatin phase
to the starch phase, decreasing the Tg of gelatin. Among all films, starch
and gelatin showed closer peak position in the S/G-RO films, which
indicated better compatibility between starch and gelatin. This was
possibly due to that RO could facilitate some interactions between the
starch and gelatin at the molecular level to a greater extent.
3.6. Surface hydrophobicity
Surface hydrophobicity plays an important role in food packaging,
characterized by the water contact angle. The DCAs of all S/G films
within 90 s were shown in Fig. 7. The water contact angles of all films
decreased with prolonged time, which was attributed to the reor­
ientation of polar groups on the film surface. The S/G-lipid films
exhibited higher final contact angle compared with the control, showing
a significant improvement in hydrophobicity. This demonstrated that
lipid particles were present on the film surface and increased their hy­
drophobicity. Among all the S/G films, the OS/G-RO film showed the
strongest hydrophobicity. High hydrophobic surface can be built by
rougher surface, which are caused by the aggregation of lipids and their
following crystallization during film-forming process [17]. Among all
the HP/G films, HP/G-RO film also exhibited the highest contact angle
and only reduced by 3.7◦ within 90 s after the water drop. Particularly,
Y. Cheng et al.
Fig. 7. Dynamic water contact angles of OS/G-lipid (a) and HP/G-lipid (b) edible films within 90 s.
the final contact angle of the HP/G-RO film is greater than 90◦ , exhib­
iting the characteristics of a hydrophobic film surface. This indicates
that the formation of continuous hydrophobic layer on the surface of
HP/G-RO film, which supported the SEM analysis.
In addition, the DCAs of the films were influenced by the type of
starch. By comparison, the initial contact angles of OS/G-lipid films
were higher than those of HP/G-lipid films, which was attributed to the
higher surface roughness in OS/G-lipid films. However, The OS/G-RO
film exhibited a rapider drop (32.8◦ ) and lower ultimate value (72.6◦ )
in the water contact angle after the water drop than HP/G-RO film,
which could be attributed to that the RO was mainly present on the
surface of the OS/G-RO film as dispersed lipid particles.
3.7. Water vapor barrier property
Water vapor barrier properties of edible films plays an important role
in the application of these films in food packaging to control the transfer
of moisture and extend the shelf life of food products. A lower WVP
value indicates a stronger water vapor barrier property. As shown in
Table 2, the addition of lipids in S/G films caused significant decrease in
WVP values (p < 0.05). The dispersion of lipids in the film matrix could
lead to discontinuities in the hydrophilic phase, which increased the
effective path length for water diffusion and thereby decreasing the
film's WVP. Among all the S/G-lipid films, the S/G-BW films exhibited
lower WVP values. That is to say, the S/G-BW edible films showed better
moisture barrier properties. Wax usually exhibits lower polarity due to
its higher content of non-polar long-chain fatty alcohols, higher fatty
acid esters and alkanes, which could more effectively prevent water
vapor from permeating through films [9]. RO was more effective than ST
in enhancing the water vapor barrier property of S/G films. This indi­
cated that there were other reasons related to the structural arrangement
of the lipid in the film matrix which played an important role [28]. As
commented above, the RO tended to migrate and aggregate, achieving
successive large lipid layers which exhibited stronger water vapor bar­
rier ability.
3.8. Water resistance properties
The WS and SD of edible films was often used to measure the water
resistance. The high water resistance of edible films was essential to
maintain the integrity of the package in the practical application.
Table 2 listed the WS and SD values obtained for all S/G films. The S/G
films without the lipids exhibited the highest WS and SD values, indi­
cating the lowest water resistance. Starch and gelatin could combine
with water molecules to form hydrogen bonds due to their hydrophilic
nature, which promoted their dissolution in water [29]. The
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International Journal of Biological Macromolecules 185 (2021) 1005–1014
incorporation of lipids significantly enhanced the water resistance of S/
G films. The lower WS values of S/G-lipid films was mainly attributed by
the fact that the incorporation of lipids could decrease the hydrophilicity
of films. The non-polar components of lipids could also interact with S/G
matrix, reducing the binding sites of water molecules [22]. Moreover,
the addition of lipids also promoted the plasticization of starch to form a
denser film network structure, which was positive for reducing water
absorption, thereby weakening film swelling. Among all the S/G-lipids
films, HP/G-BW film showed the lowest WS and SD. Compared with
HP, OS introduces a large number of hydrophilic carboxyl and carbonyl
groups, which tended to contact with water and promote the dissolution
and swelling of the OS/G films.
3.9. Mechanical properties
Mechanical properties are another important factor to evaluate
edible films for food packaging. Packaging materials require appropriate
mechanical properties to withstand various stresses in food packaging,
storage and transportation. The mechanical properties of the S/G films
with different lipids were shown in Fig. 8. The TS of the S/G films was
significantly improved by the addition of lipids (p < 0.05). The TS of the
OS/G film without lipids was 2.23 MPa, which increased about 44.6%,
23.5%, and 24.4% for the OS/G film containing RO, ST, and BW,
respectively (Fig. 8a). This results indicated that the lipids addition into
the film matrix could build a strong film network structure. Because the
partial stronger polymer-polymer interactions may be replaced by
weaker polymer-lipid interactions in the film-forming network, the
addition of lipids commonly decreased the TS of edible films [30,31].
Nevertheless, the TS of S/G films in our research all notably improved
with the incorporation of lipids. These different test results should be
attributed to the difference in film-forming materials and process, as
well as the lipid type. Zhang et al. [8] reported the comparable results
that the addition of lipids increased the TS of agar/maltodextrin films.
After adding lipids, the starch granules were melted to a greater extent,
resulting in the release of hydrophilic starch chains to the film system.
These starch chains were free to form chemical and/or physical cross­
linking with gelatin to enhance the strength of the film structure.
Moreover, the addition of lipids increased the film's crystallinity, which
also favored the improvement of the TS [32].
Lipids also acted as a plasticizing agent as revealed by SEM, which
could increase the flexibility of the polymer chains and lead to a higher
EAB. However, this mechanism didn't occur for the OS/G-RO films. In
fact, the EAB of OS/G-RO film was even lower than that of the control
film. This different effect was related to the type of starch and lipids. The
oxidation of starch can cause the formation of dialdehyde, which pro­
moted the crosslinking capacity of the polymer chains and thereby
Y. Cheng et al.
Fig. 8. Tensile strength (a), elongation at break (b) and Young's modulus (c) of S/G-lipid films.
improving the TS of the film [33]. Additionally, RO was more weakly
bound with amylose than ST and BW due to the lower saturation, which
promoted cross-linker effect between starch and gelatin [21]. These
effects resulted in a higher value of TS and lower value of EAB for the
OS/G-RO film. The YM of the HP/G films were notably improved when
added with any of these lipids, while the addition of ST and BW into OS/
G film slightly reduced its YM value. The RO addition significantly
improved the YM of OS/G film, leading to stiffer and more resistant
films. Based on the results, OS/G-RO film was the most rigid one.
4. Conclusion
The physicochemical properties of S/G films were investigated to be
significantly affected by the incorporation of three lipids. The film-
forming capacity of S/G and the properties of the films were notably
improved by the addition of lipids, which was significantly influenced
by the type of lipid. Rheological analysis determined that the addition of
lipids decreased the G′ and |η*| values of S/G blends. ATR-FTIR indi­
cated that gradually weakening molecular interactions in S/G-BW and
S/G-ST films. XRD showed an increased crystallinity with the addition of
lipids. Compared with BW and ST, RO addition promoted starch plas­
ticization to a greater extent determined by the SEM analysis. DMA
indicated that starch and gelatin exhibited better compatibility in S/G-
lipid films, especially for the S/G-RO films. The TS and EAB of the S/
G films were simultaneously improved with the addition of lipids. Lipids
had positive effects on the water resistance, water vapor barrier prop­
erties and surface hydrophobicity of the films. RO was more effective in
improving TS and surface hydrophobicity. The strongest water vapor
barrier and water resistance properties of film were obtained by the
addition of BW. HP/SG-lipid films showed lower WVP, WS, SD and
higher EAB than OS/SG-lipid films. These results highlight the potential
of the S/G films with lipids for food packaging applications with optimal
hydrophobicity, mechanical and water vapor barrier performance.
Declaration of competing interest
The authors declare that they have no conflict of interest in the
publication of this article.
Acknowledgments
This work was supported by Great Innovation Program of Agricul­
tural Application Technology in Shandong Province, the Key Technol­
ogy Research and Development Program of Shandong province
(2019GNC106035), Natural Science Foundation of Shandong Province
(ZR2020QC222), Natural Science Foundation of Tianjin City
(18JCQNJC84200), and Funds of Shandong “Double Tops” Program.
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International Journal of Biological Macromolecules 185 (2021) 1005–1014
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