polymers

Article
Preparation and Performance Characterization of a Composite
Film Based on Corn Starch, κ-Carrageenan, and Ethanol Extract
of Onion Skin
Cuntang Wang 1,2, *, Yueyi Lu 1 , Ziyu Li 1 , Xuanzhe An 1 , Zengming Gao 1 and Shengxin Tian 1

1 College of Food and Bioengineering, Qiqihar University, Qiqihar 161006, China;

[email protected] (Y.L.); [email protected] (Z.L.); [email protected] (X.A.);
[email protected] (Z.G.); [email protected] (S.T.)
2 College of Food Science, Northeast Agricultural University, Harbin 150030, China
* Correspondence: [email protected]

Abstract: Using corn starch (CS) and κ-carrageenan(κC) as the raw material and active composite,
respectively, films containing different concentrations of ethanol extract of onion skin were prepared.
The effects of different concentrations of ethanol extract of onion skin (EEOS) on the physicochemical
properties, as well as the antioxidant and antibacterial properties, of CS/κC films were also discussed.
The addition of ethanol extract of onion skin inhibited the recrystallization of starch molecules in the
composite films. It affected the microstructure of the composite films. The color of the composite
films was deepened, the brightness was reduced, and the opacity was increased. Water vapor
permeability increased, tensile strength decreased, and elongation at the break increased. The glass-
transition temperature decreased. The clearance of DPPH radicals and ABTS cation radicals increased.
Moreover, when the concentration of EEOS was 3%, the antioxidant effect of the films on oil was
greatly improved and could effectively inhibit Staphylococcus aureus and Escherichia coli. The above
results showed that adding ethanol extract of onion skin improved the physicochemical properties
Citation: Wang, C.; Lu, Y.; Li, Z.; An,
and biological activities of the CS/κC composite films, so CS/κC/EEOS composite films can be used
X.; Gao, Z.; Tian, S. Preparation and
Performance Characterization of a
as an active packaging material to extend food shelf-life. These results can provide a theoretical
Composite Film Based on Corn basis for the production and application of corn starch/κ-carrageenan/ethanol extract of onion skin
Starch, κ-Carrageenan, and Ethanol composite films.
Extract of Onion Skin. Polymers 2022,
14, 2986. https://doi.org/10.3390/ Keywords: corn starch; κ-carrageenan; ethanol extract of onion skin; active film; physicochemical
polym14152986 properties; biological activity
Academic Editor: Alexey V. Lyulin

Received: 4 July 2022

Accepted: 20 July 2022 1. Introduction
Published: 23 July 2022
Traditional petroleum-based plastic films are widely used as packaging materials
Publisher’s Note: MDPI stays neutral for their low-density, low-cost, and good molding properties [1]. However, the slow
with regard to jurisdictional claims in biodegradation of food plastic packaging materials has caused global increases in plastic
published maps and institutional affil- waste [2]. Thus, the development of eco-friendly and safe packaging materials has emerged
iations. as an area of interest [3]. Concerns about environmental issues associated with the large
disposal of nondegradable plastics and diminishing supplies from oil sources drive efforts
to investigate sustainable materials that are degradable and are from renewable sources [4].
When considering the sustainability of the ecosystem, more people prefer to transfer their
Copyright: © 2022 by the authors.
interest to degradable materials [5]. Biopolymer materials have attracted more and more
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attention due to their excellent properties, such as biodegradability, safety, biocompatibility,
This article is an open access article
and renewability [6]. In recent years, biological materials such as polysaccharides, proteins,
distributed under the terms and
conditions of the Creative Commons
and lipids have been highly focused on [7]. Biodegradable films can be developed using
Attribution (CC BY) license (https://
proteins, carbohydrates, lipids, or compounds [2]. Moreover, films based on biodegradable
creativecommons.org/licenses/by/ materials are often incorporated with other bioactive compound blends, such as plant
4.0/). extracts, to satisfy the changing preferences and expectations of modern consumers [8].

Polymers 2022, 14, 2986. https://doi.org/10.3390/polym14152986 https://www.mdpi.com/journal/polymers

Polymers 2022, 14, 2986 2 of 19

Among natural and renewable resources, starch, as a significant raw material, has attracted
extensive attention due to its low production cost, availability, and biodegradability, as well
as its odorless, colorless, and nontoxic biodegradable film qualities and its ease of use in
film production [9]. Among biopolymer materials, corn starch is considered one of the most
promising plastic alternatives since it is abundant, inexpensive, biodegradable, edible, and
presents good film-forming ability [10]. Although starch-based materials have good oxygen
barrier properties, compared with traditional plastics, the use of corn starch to produce
biodegradable plastics has disadvantages, such as poor moisture resistance and poor
mechanical properties, and its application is limited [2]. To overcome these shortcomings,
mixing starch with other natural biopolymers to form composites has received much
attention [11].
Carrageenan is one of the algal colloids among several polysaccharides extracted from
seaweed. It consists of alternating disaccharide units of β-(1-4)-3,6-anhydrous-D-galactose
and α-(1-3)-D-galactose. It is commercially classified into three categories, κ (kappa), ί
(iota), and λ (lambda), according to differences in the content of the sulfate group and
its position in the structure [12]. Carrageenan is increasingly regarded as a promising
renewable biomaterial with excellent film-forming ability and has great potential as a
substitute for traditional synthetic plastics. In recent decades, Carrageenan has been widely
used in edible films and coatings [13]. However, carrageenan films have limitations. The
κ-carrageenan monomer selected by the research institute has only one sulfate group and
has good film-forming properties but poor mechanical properties. In this experiment,
corn starch was mixed with κ -carrageenan and could form a film with good mechanical
properties [12]. To expand the scope of application, a large number of studies have been
carried out on how to improve the performances of films [12]. One way to improve the
performance of thin films is to add active compounds. Natural compounds are preferred
instead of synthetic additives. Natural extracts rich in polyphenols are an effective way
to prevent lipid oxidation in food and can be used as a functional additive. Some studies
on biopolymer films based on natural antioxidants have been reported [14]. Due to their
superior biodegradable, biocompatible, and economic advantages, composite films mixed with
natural extract materials have been widely studied and applied [13].
The onion (an Allium of the Liliaceae family) is one of the most essential and widely eaten
vegetables globally. It is the second most cultivated crop after tomatoes. However, about
500,000 tons of inedible onion waste are produced every year [15]. Onion waste is mainly
made up of onion skins, which are rich in bioactive phenolic compounds. Polyphenols are
a kind of natural antioxidant, and their groups include flavonoids and anthocyanins [16].
Purple onions obtain their color from anthocyanins in their epidermal cells. In flavonoids,
anthocyanins are considered the most biologically active compounds with a high antiox-
idant performance [17]. Although onion skins are rich in fiber and flavonoids, the dry,
outer layer of onion skin is considered waste and is discarded, resulting in a significant
loss of phenolic compounds [18]. Phenolic compounds can act as free radical scavengers
and metal ion chelators to inhibit lipid oxidation [16]. Therefore, onion skin extract can be
added to films as antioxidant material to prepare food-active packaging and to develop a
biodegradable antioxidant film.
However, there are few reports on preparing composite films by blending onion
skin extract with corn starch and κ-carrageenan. In this study, ethanol extract of onion
skin are added to a composite film to improve its physical and chemical properties. The
effects of different contents of EEOS on the properties of CS/κC films are also evaluated to
select a good food-packaging film. The characterization of CS/κC/EEOS composite films
investigates the effects of EEOS on film properties, including Fourier-transform infrared
spectroscopy, crystal structure, morphology, optical properties, mechanical properties,
water vapor permeability, thermodynamic property, antioxidant activity, and antibacterial
activity. We hope to provide environment-friendly film materials for food packaging.
Therefore, the purpose of this study is to prepare environmental-friendly and functional
Polymers 2022, 14, 2986 3 of 19

CS/κC/EEOS composite films with different concentrations of ethanol extract of onion

skin as raw materials.

2. Materials and Methods

2.1. Materials and Reagents
Purple onion skin was obtained from the Liu yuan Market (Qiqihar, China); corn
starch, was obtained from Heilongjiang YuFeng Corn Development Co., Ltd. (Heilongjiang,
China); food-grade κ-carrageenan was obtained from Qingdao DeHui Marine Biotech-
nology Co., Ltd. (Qingdao, China); beef paste and tryptone (biochemical reagent) were
obtained from Beijing AoBoxing Biotechnology Co., Ltd. (Beijing, China); Staphylococcus
aureus (ATCC29213) and Escherichia coli (ATCC25922) were obtained from the College
of Food and Bioengineering of Qiqihar University (Qiqihar, China); and 2,2-Diphenyl-1-
picrylhydrazyl (DPPH) and 2,20 -azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)
were obtained from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). All other chemical
reagents (analytically pure) were obtained from Tianjin KaiTong Chemical Reagents Co.,
Ltd. (Tianjin, China).

2.2. Preparation of Ethanol Extract of Onion Skin

The purple onion skins were washed, blasted in an air oven (GZX-9146MBE, Shanghai
BoXun Industrial Co., Ltd., Shanghai, China) to dry them, crushed with a shredder (GX-220,
Zhejiang GaoXin Industrial and Trade Co., Ltd., Yongkang, China), and sifted. A specific
mass of purple onion skin powder was weighed, and ethanol solution with the 70% volume
fraction was added in a ratio of 1:2 (W:V). The solution was extracted at 25 ◦ C for 2 h, then
the leached liquor was centrifuged at 6000 r/min for 10 min, and the supernatant was
obtained for standby application. After centrifugation, the sediment was extracted and
centrifuged again. Two centrifugates of supernatants were combined. The supernatant
was transferred to a rotary evaporator (2L-ARE, Shanghai HaoZhuang Instrument Co.,
Ltd., Shanghai, China) and concentrated at 50 ◦ C. Then, the concentrated solution was
freeze-dried with a vacuum freeze-dryer (2.5 L freezer dryer, Labconco company, Kansas
City, MO, USA) to obtain the EEOS. The EEOS was stored in a refrigerator at −20 ◦ C until
ready for use.

2.3. Preparation of CS/κC/EEOS Complex Films

A certain amount of corn starch was added into distilled water to prepare a suspension
of corn starch with a concentration of 3% and was gelatinized at 90 ◦ C for 30 min. An
amount of 30% κ-carrageenan (W/W: κ-carrageenan mass/starch dry-base mass) was
added, and we continued to heat and stir at 90 ◦ C for 30 min. Leaving one control, amounts
of 1%, 3%, and 5% ethanol extract of purple onion skin (W/W: extract weight/starch
dry-base weight) were added and, respectively, called CS/κC films, CS/κC/EEOS I films,
CS/κC/EEOS II films, and CS/κC/EEOS III films. We continued to stir for 30 min, added
25% glycerin (W/W: glycerin mass/starch dry-base mass), and continued to heat and stir
at 90 ◦ C for 30 min, using hot water to complement the evaporated water. The solution
was ultrasonicated at 80 ◦ C for 30 min to remove bubbles, and the composite film solutions
were prepared. A polyethylene ring with a diameter of 120 mm was fixed on a glass plate
covered with release paper. An amount of 25 g of film solution was cast in the polyethylene
ring and fixed for 15 min. The samples were placed in an air-blast drying oven and dried at
40 ◦ C for 12 h. Then, the films were uncovered. The films were placed in a dryer at 25 ◦ C
with a relative humidity of 56.8% (NaBr saturated solution) and were balanced for 72 h to
determine the parameters of the films.

2.4. Characterization
2.4.1. Determination of Fourier-Transform Infrared Spectroscopy (FTIR)
Infrared spectra were measured with a Fourier-transform infrared spectrometer
(Spectrum-100, PerkinElmer company, Waltham, MA, USA). After being balanced and
Polymers 2022, 14, 2986 4 of 19

dried, the films were placed on the ATR accessory. The test temperature was 25 ◦ C, the
wavenumber was 4000–650 cm−1 , the resolution was 4 cm−1 , and the number of scans
was 32.

2.4.2. X-ray Diffraction Analysis (XRD)

The samples were analyzed with an XRD instrument (Rint-2000, Neology Co., Ltd.,
Tokyo, Japan). A Cu target and a graphite monochromator were used. The test parameters
were as follows: the voltage was 45 kV, the current was 20 mA, the scanning range was
5~80◦ (2θ), and scanning rate was 2 (◦ )/min.

2.5. Morphology and Optical Properties

2.5.1. Scanning Electron Microscope (SEM)
Each composite film sample was cut into a brittle rectangle of 4 cm × 6 cm and was
broken using liquid nitrogen. A scanning electron microscope (S-4300, Hitachi, Japan) was
used for scanning the microstructure. The scanning voltage was 2.00 KV, and the current
was 64.0 µA. The surface and cross-section structures of the films were photographed
and observed.

2.5.2. Optical Properties

For the measure of color and luster, the a * value (red and green), b * value (yellow
and blue), and L * value (brightness) of the film samples were measured with a visible
spectrophotometer (UPG-722, Beijing YouPu General Technology Co., Ltd., Beijing, China).
Before measurement, it was calibrated with a standard board (a * = −5.80, b * = 9.25, and
L * = 103.98). Each sample was measured three times, and the results were averaged.
For the measure of opaqueness, according to Sukhija et al. [19], each film sample was
cut into a 10 mm × 45 mm rectangle, which was closely attached to the inner wall of an
empty colorimeter. The empty colorimeter was used as a reference, and the absorbance
values of the samples were measured at 600 nm. The opacity was calculated according to
Equation (1). Each sample was measured three times, and the average value was taken.

A600
Opaqueness = (1)
X
In the formula, A600 is the absorbance value at a wavelength of 600 nm, and X is the
film thickness in mm.
For appearance, the composite film was covered on A4 white paper, and an image
was captured of the composite film.

2.6. Mechanical and Barrier Properties

2.6.1. Determination of Mechanical Properties
The mechanical properties of the films were slightly modified by referring to Mu et al. [20].
The films were cut into 6 cm × 2 cm rectangles, the test speed was 2 mm/s, and the initial
clamping distance was 20 mm. The tensile strength (MPa) and elongation at the break
(%) of the film samples were measured with a texture tester (TA. XT plus, Stable Micro
System company, Surrey, UK). Each sample was measured three times, and the results were
averaged. The mechanical properties of the carrageenan films were calculated according to
Equations (2) and (3):
Fmax
TS = (2)
S
In the formula, TS is the tensile strength in MPa, Fmax is the maximum load when the
film breaks in N, and S is the film cross-sectional area in mm2 .

L1 − L0
EB% = × 100% (3)
L0
Polymers 2022, 14, 2986 5 of 19

In the formula, EB is the elongation at the break as a percentage, L1 is the length of the
film after stretching in mm, and L0 is the initial length of the film at 20 mm.

2.6.2. Determination of Water Vapor Transmittance (WVP)

Ten grams of anhydrous CaCl2 was placed in a blast-drying oven, dried at 110 ◦ C
for 2 h, and placed in a 35 mm × 90 mm weighing bottle. The prepared film sample was
covered over the weighing bottle’s mouth, sealed, and placed in a dryer with distilled
water at the bottom. The weight of the measuring bottle was measured every 24 h and
continuously for 10 days. Each sample was measured three times, and the results were
averaged. The water vapor permeability of the film could be calculated according to
Equation (4):
W X
WVP = × (4)
t × A ∆P
In the formula, WVP is the water vapor permeability (g·mm/m2 ·d·kPa), W is the
the total weight of the measuring bottle after film sealing (g), t is the time (d), A is the
permeable film area (m2 ), X is the film thickness (mm), and ∆P is the vapor pressure
difference between the two sides of the film (1.583 kPa).

2.7. Differential Scanning Calorimetry (DSC)

Differential scanning calorimetry was used for the thermal stability analysis. About
4 mg of film samples dried with a dryer were taken and sealed in an aluminum crucible,
using an empty aluminum crucible as a reference. Nitrogen was the protective gas, with
a flow rate of 20 mL/min. The heating rate was 10 ◦ C/min, and differential scanning
calorimetry was performed in the temperature range from 20 ◦ C to 250 ◦ C.

2.8. Determination of Total Phenol Content (TPC)

The total phenol content of the composite film was determined using the Folin-phenol
colorimetric method. A total of 125 mg film sample was immersed in 15 mL distilled water
for 24 h to obtain a film immersion solution. Amounts of 0.1 mL film immersion solution,
7 mL distilled water, and 0.5 mL Folin-Phenol were successively added to a 50 mL conical
flask and gently shaken. After 8 min of rest, 1.5 mL 10 wt% sodium carbonate solution and
0.9 mL of distilled water were added successively, and the composite solution was put into
a dark room to avoid light for 2 h. Then, the absorbance of the mixture was measured at 765
nm with an ultraviolet spectrophotometer. A series of aqueous gallic acid solutions with
concentrations between 0 and 15 µg/mL were prepared, and the absorbance at 765 nm
was measured according to the above steps. The standard curve was drawn with the
concentration of aqueous gallic acid as the abscissa and the absorbance at 765 nm as the
ordinate. The equation y = 0.117x + 0.0171 (R2 = 0.9995) was obtained. According to this
equation, the total phenol content was calculated using the absorbance of the composite
film immersion solution at 765 nm. The total phenolic content of the sample was expressed
as the mg gallic acid equivalent (GAE) per gram of dry matter (DW), or GAE mg/DW g.
Each sample was measured three times, and the results were averaged.

2.9. Antioxidant Properties

2.9.1. Determination of Antioxidant Capacity In Vitro
A total of 10 mg film sample was mixed with a methanol DPPH solution (0.2 mM;
1.5 mL). The mixture was put into a dark chamber for reaction and kept away from light
for 30 min at indoor temperature. Then, the absorbance of the mixture was measured
at 517 nm with a UV-vis spectrophotometer, using the DPPH methanol solution as the
control [21]. Each sample was measured three times, and the results were averaged. The
equation used for calculating the free radical clearance rate (Equation (5)) was as follows:

AC − As
k= × 100% (5)
AC
Polymers 2022, 14, 2986 6 of 19

In the formula, k is the free-radical-scavenging ability (%), AC is the the absorbance of

control group, and AS is the absorbance of sample
A total of 10 mg film sample was mixed with ABTS free radical working solution and
kept from light at room temperature for 6 min. Using a UV-vis spectrophotometer, the
absorbance of the mixture was determined at 734 nm. An acetic acid buffer solution was
used instead of an extract sample solution as a blank control. Each sample was measured
three times, and the results were averaged. The free radical clearance rate was calculated
according to Equation (5).

2.9.2. Determination of Peroxide Value (POV)

An amount of 5 g solid lard was wrapped in the film, and the film was heat-sealed and
placed in a blast-drying oven at 60 ◦ C to accelerate oxidation. The samples were sampled
at every 24 h interval and measured continuously for 7 d. The POV value was determined
according to the method of Gao et al. [22]. Each sample was measured three times, and the
results were averaged.

2.10. Determination of Antibacterial Properties

Cultured Escherichia coli and Staphylococcus aureus strains were diluted ten times with
sterile normal saline as the initial bacterial solution. The thin-film samples were cut into
discs with diameters of 7 mm with a sampler. After being sterilized with ultraviolet
radiation, they were placed in a culture medium containing 0.1 mL target bacteria solution
and incubated in a constant-temperature incubator at 37 ± 1 ◦ C for 24 h. Use vernier
calipers to measure the diameters of the membrane inhibition zones, the antibacterial
activity of the film was judged by the size of the antibacterial area. Each sample was
measured three times, and the results were averaged.

2.11. Data Processing

The experimental results were expressed as average values ± standard deviation.
Duncan’s multiple range test in SPSS 26 software (SPSS Inc., Chicago, IL, USA) was used to
analyze the significance of the difference, and Origin 2019 software (Microsoft, WSU, USA)
was used to plot.

3. Results
3.1. FTIR
The infrared spectra refer to the absorption bands formed by molecules selectively
absorbing specific frequencies under infrared irradiation. FTIR spectroscopy can not only
measure the functional groups of each component in the film but also can be used to
detect the molecular interactions between the components of the film [23]. The interactions
between EEOS and the CS/κC composite films were further analyzed by Fourier-transform
infrared spectroscopy (FTIR). The FTIR spectra of CS/κC/EEOS composite films in the
range of 4000~520 cm−1 are shown in Figure 1.
For the CS/κC films, the starch contained many hydroxyl groups. There was a wide
band at 3304 cm−1 , where the peak appeared as O-H vibration stretching, and the shifting
of the O-H stretching peak suggested interaction between the polymer and additives
via H-bonding [24]; the peak at 2926 cm−1 was a C-H stretching vibration peak. The
absorption peak at 1645 cm−1 was attributed to the tight binding between starch and
water. Meanwhile, it was shown that the characteristic peak is related to the crystallinity
of starch, and the decrease in crystallinity leads to an increase in the strength of the
characteristic peak [25]. The C-H and C-O stretching of starch were respectively observed
at 1365 cm−1 and 1149 cm−1 . Glucose pyranose ring vibration was seen at 993–1079 cm−1 .
For carrageenan, the stretching vibration peak of the O-H bond appeared at 3304 cm−1 .
The stretching vibration peak of the C-H bond occurred at 2926 cm−1 . The stretching
vibration peak of the C=O bond appeared at 1645 cm−1 . The peak at 1365 cm−1 was from
the in-plane-bending vibration of the O-H bond. The absorption peak at 1008 cm−1 came
 Polymers 2022, 14, 2986 7 of 19

Polymers 2022, 14, x FOR PEER REVIEW 7 of 20

from the stretching vibration of the C-O bond. The characteristic peaks at 1149, 1079, 927,
and 854 cm−1 , respectively, corresponded to the sulfate; glycosidic bond; 3, 6-anhydrous
galactose; and galactose 4-sulfuric acid in the κ-carrageenan structure [12].

Figure 1. FTIR spectra of CS/кC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III
Figure 1. FTIR spectra
composite films.of CS/кC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III composite
films.
After adding EEOS, the infrared spectra curves of the composite films did not change
significantly,
For the CS/κC films, indicating that the
the starch addition
contained manyof EEOS did not
hydroxyl causeThere
groups. structure
was achange
wide in the
band atcomposite
3304 cm ,films
−1 where and
thethat
peakthere was good
appeared as O-Hcompatibility between the
vibration stretching, andtwo.
the The absorption
shifting
peaks
of the O-H at 2926 and
stretching peak1008 cm−1 , respectively,
suggested moved to
interaction between the2929 and 1014
polymer andcm
−1 , which may be
additives via
due to
H-bonding thethe
[24]; increase
peak atin2926
chemical bond
cm−1 was forcestretching
a C-H and stretching vibration,
vibration peak. making the absorption
The absorption
peak atpeak
1645move
cm−1to wasa higher wavenumber.
attributed to the Similar changesbetween
tight binding in band location and peak
starch and water.strength
Meanwhile, it was shown that the characteristic peak is related to the crystallinity of [26].
were observed when food anthocyanin was added to starch/polyvinyl alcohol films
starch, These results
and the indicated
decrease that the interactions
in crystallinity leads to between components
an increase can be identified
in the strength of the with
FTIR. The increase in band strength at 3304, 2926, and 1008 cm −1 of the composite films
characteristic peak [25]. The C-H and C-O stretching of starch were respectively observed
at 1365was
cm−1caused
and 1149 by hydrogen bonding
cm−1. Glucose between
pyranose ringthe composite
vibration wasfilms
seenand EEOS [27].
at 993–1079 cm−1.
For carrageenan, the stretching vibration peak of the O-H bond appeared at 3304 cm−1.
3.2. XRD Analysis
The stretching vibration peak of the C-H bond occurred at 2926 cm−1. The stretching
Theofcrystallinity
vibration peak of polymers
the C = O bond canat
appeared be1645
analyzed
cm−1. with XRD. at
The peak The X-ray
1365 cmdiffraction
−1 was from patterns
of a CS/κC/EEOS composite film are shown in Figure
the in-plane-bending vibration of the O-H bond. The absorption 2. peak at 1008 cm−1 came
CS/κC films had three diffraction peaks at 2θ = 17.7 ◦ , 2θ = 19.7◦ , and 2θ = 22.1◦ . The
from the stretching vibration of the C-O bond. The characteristic peaks at 1149, 1079, 927,
and 854diffraction peaks at 2θ
cm−1, respectively, = 17.7◦ and to
corresponded 19.7
thewere dueglycosidic
sulfate; to the recrystallization of amylose and
bond; 3, 6-anhydrous
amylopectin during the storage of the starch composite films [28]. After adding the EEOS,
galactose; and galactose 4-sulfuric acid in the κ-carrageenan structure [12].
the diffraction peak at 2θ = 17.7◦ decreased with the addition of EEOS. The diffraction
After adding EEOS,◦the infrared spectra curves of the composite films did not change
peak at 2θ = 22.1 increased with the addition of EEOS. However, when the amount of
significantly, indicating that the addition of EEOS did not cause structure change in the
EEOS added reached 5%, the diffraction peak here disappeared, and the composite films
composite films and that there was good compatibility between the two. The absorption
only showed an obvious peak at 19.7◦ . The addition of EEOS affected the crystal structure
peaks at 2926 and 1008 cm−1, respectively, moved to 2929 and 1014 cm−1, which may be
of the composite film, which may be because of the interaction between the polyphenols
due to the increase in chemical bond force and stretching vibration, making the absorption
contained in EEOS and the starch molecules, affecting the order of the starch molecular
peak move to a higher wavenumber. Similar changes in band location and peak strength
structure and preventing the recrystallization of starch. This is similar to the research results
were observed when food anthocyanin was added to starch/polyvinyl alcohol films [26].
of Ren [11]. The crystallinity of the starch film comes from the crystallization of amylose
These results indicated that the interactions between components can be identified with
and amylopectin. The crystallization rate of amylopectin is slow and occurs during storage,
FTIR. The increase in band strength at 3304, 2926, and 1008 cm−1 of the composite films
so the recrystallization of starch composite films occurs during storage. The molecular
was caused by hydrogen
interaction betweenbonding
EEOS andbetween the composite
the CS/κC films the
matrix affects andrecrystallization
EEOS [27]. of the starch.
Sun et al. made a similar discovery, finding that the interaction between anthocyanins
3.2. XRD Analysis
and polymers and the promotion of the spatial reconfiguration of polymer chains affected
The
thecrystallinity of polymers
lattice structure can chloride
of potassium be analyzed
[12]. with XRD. between
Interaction The X-ray diffraction
polymers causes the
patternsfine
of adispersion
CS/κC/EEOS composite film are shown in Figure 2.
of bulky polyphenolic compounds, which physically prevents close contact
 Polymers 2022, 14, 2986 8 of 19

Polymers 2022, 14, x FOR PEER REVIEW 8 of 20

of the polymers and prevents crystallization [29]. The results showed that the addition
of plant extracts had a good effect on maintaining the good mechanical properties of the
composite films during storage.

Figure 2. XRD spectra of CS/кC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III
Figure 2. XRD spectra of CS/кC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III composite
composite films.
films.
3.3. SEM
CS/κC films had three diffraction peaks at 2θ = 17.7°, 2θ = 19.7°, and 2θ = 22.1°. The
SEM can observe the microstructure of the films and the interactions and influences
diffraction peaks at 2θ = 17.7° and 19.7 were due to the recrystallization of amylose and
between the films and additives. The surface and cross-section microstructures of the
amylopectin during the storage of the starch composite films [28]. After adding the EEOS,
CS/κC/EEOS composite films are shown in Figure 3.
the diffraction
As canpeak at 2θfrom
be seen = 17.7° decreased
Figure 3A–D, the withCS/κC
the addition of EEOS.
films without The were
EEOS diffraction
relatively
peakcompact
at 2θ = 22.1°
and smooth, indicating that there was good compatibility between corn of
increased with the addition of EEOS. However, when the amount starch
EEOS andadded reached 5%,
carrageenan. With thethe
diffraction
increase in peak
EEOS hereaddition
disappeared,
from 1% andtothe5%,composite
the surfaces filmsof the
onlycomposite
showed anfilms obvious peak atpresented
gradually 19.7°. The aaddition
convex of EEOS affected
structure, and thethepresence
crystal structure
of insoluble
of the composite
particles couldfilm, which maywhich
be observed, be because of the
indicated interaction
that a small amountbetween of the
EEOS polyphenols
was insoluble
contained in EEOS and the starch molecules, affecting the order
in the films, making the film surfaces uneven. Gasti et al. [30] also observed of the starch molecular
a similar
structure and preventing
phenomenon. It can be theseen
recrystallization
from Figure 3a–d of starch.
that theThis is similar to of
cross-sections the
theresearch
films were
results of Rento[11].
identical the The crystallinity
surface structure.ofThethe CS/κC
starch film
filmcomes from the
had a more crystallization
compact structure of and a
amylose and amylopectin. The crystallization rate of amylopectin
smooth cross-section. With the addition of EEOS, the section began to gradually is slow and occurs become
during storage,
rough. Whensothe thecontent
recrystallization of starch
of EEOS reached 5%, composite films occurs
the cross-sections during
of the films storage.
became coarser,
The which
molecular interaction
corresponded between EEOS
to the phenomenon of theand thestructure.
surface CS/κC Thesematrixresults
affects
showed the that
recrystallization
EEOS was evenly of the starch. Sun
distributed et al.
at low made a similar
concentrations, discovery,
but the finding thatwas
effect of uniformity themore
interaction between anthocyanins and polymers and the promotion
significant at higher concentrations. Due to the higher extract concentration, the polymer of the spatial
reconfiguration
network was of destroyed,
polymer chains affected
and the the lattice
structure structure
was coarser andof potassium
less compact.chloride [12].
Furthermore,
Interaction
the spatialbetween
structure polymers causes the
of the composite film fine dispersion
was affected. of bulky
Similarly, polyphenolic
reports of novel films
compounds,
combiningwhichstarchphysically
with polyvinylprevents
alcoholclose
and contact of the polymers
food anthocyanin have alsoandshownprevents
that a low
crystallization
content of [29].
PSPEThe results
in films hadshowed
a compactthat structure.
the addition of plant extracts
In contrast, excessive had a good
PSPE hadeffect
a greater
on maintaining
impact on the theuniformity
good mechanical properties
of the films of the composite
and showed films [26].
a rough surface during storage.

3.3. SEM
SEM can observe the microstructure of the films and the interactions and influences
between the films and additives. The surface and cross-section microstructures of the
CS/κC/EEOS composite films are shown in Figure 3.
Polymers 2022,
Polymers 2022, 14,
14, 2986
x FOR PEER REVIEW 99 of
of 19
20

3. SEM micrographs of the surfaces

Figure 3. surfaces and
and cross-sections
cross-sections of
of composite
composite films.
films. (A–D) are the
(a–d) are
surfaces, and (a–d) are the
the cross-sections
cross-sections of
of composite
composite films.
films.

3.4. Optical Properties

As can be seen from Figure 3A–D, the CS/κC films without EEOS were relatively
Color
compact andis an important
smooth, index that
indicating to measure
there was food-packaging films. between
good compatibility a *, b *, and
cornLstarch
* are
used to distinguishWith
and carrageenan. colorthedifferences.
increase in A EEOS
positive a * value
addition represents
from 1% to 5%,thethe
redness value
surfaces ofof a
the
sample, and a positive b * value represents the yellowness value
composite films gradually presented a convex structure, and the presence of insolubleof a sample [26]. CS/κC
films containing
particles could be different
observed,concentrations
which indicatedof EEOSthatwere prepared.
a small amountThe opticalwas
of EEOS properties of
insoluble
the CS/κC/EEOS composite films are shown in Table 1.
in the films, making the film surfaces uneven. Gasti et al. [30] also observed a similar
For the CS/κC/EEOS
phenomenon. It can be seen films,
fromtheFigure
a * value
3a–dincreased
that thefrom −1.22 ± 0.04
cross-sections films±were
to 17.26
of the 0.23
(p < 0.05); the b * value increased from − 0.18 ± 0.08 to 18.06 ±
identical to the surface structure. The CS/κC film had a more compact structure and aa
0.85 (p < 0.05) and showed
significant increasing trend
smooth cross-section. With the (p <addition
0.05). It indicated
of EEOS, the thatsection
the redness
beganvalue and yellowness
to gradually become
value
rough. When the content of EEOS reached 5%, the cross-sections of thetofilms
of the composite films increased. L * decreased from 90.24 ± 0.33 ± 0.91
59.88became
(p > 0.05), and the opacity of the composite films increased significantly,
coarser, which corresponded to the phenomenon of the surface structure. These results indicating that
the color of the composite films gradually deepened, while
showed that EEOS was evenly distributed at low concentrations, but the effect of the brightness and opacity
gradually decreased. The color of the composite films gradually changed from colorless
uniformity was more significant at higher concentrations. Due to the higher extract
and transparent to red and yellow. With the increase in EEOS concentration, the color
concentration, the polymer network was destroyed, and the structure was coarser and less
of the films gradually deepened, which was consistent with the images of the films. The
compact. Furthermore, the spatial structure of the composite film was affected. Similarly,
color change in the CS/κC film may be caused by EEOS. Many studies have reported color
reports of novel films combining starch with polyvinyl alcohol and food anthocyanin have
changes in biopolymer films containing plant extracts, such as chitosan films containing
also shown that a low content of PSPE in films had a compact structure. In contrast,
 excessive
surface PSPE had a greater impact on the uniformity of the films and showed a rough
[26].
3.4. Optical Properties
surface [26].
3.4. Optical Properties
Color is an important index to measure food-packaging films. a *, b *, and L * are
3.4. Optical Properties
used Color
to is an important
distinguish
3.4. Optical Properties index to measure
color differences. A positive food-packaging
a * value represents films. the
a *, redness
b *, andvalue
L * areof
aused Color is an
to distinguish
sample, and important index represents
colorbdifferences.
a positive * value to measure
A positive food-packaging
the a * value represents
yellowness films.
value asample
of athe *, redness
b *, and LCS/κC
* are
[26].value of
used Color
to is an important
distinguish index to measure
colorbdifferences. A positive food-packaging
a *were
valueprepared. films. the
represents a *, redness
b *, andvalue
L * areofof 19
Polymers 2022, 14, 2986 a sample,
films and a positive
containing different *concentrations
value represents the yellowness
of EEOS value ofThe a sample
optical[26]. CS/κC
properties10
used
aof to distinguish
sample, and a positivecolorbdifferences.
*concentrations
value A positive
represents a * value represents
theinyellowness value ofThe athe redness value
sample of
films
thecontaining
CS/κC/EEOS different
composite films of EEOS
are shown were1.prepared.
Table optical[26]. CS/κC
properties
afilms
sample, and a positive b * value represents the yellowness value of a sample [26]. CS/κC
of thecontaining
CS/κC/EEOS different
compositeconcentrations of EEOS
films are shown were 1.
in Table prepared. The optical properties
films
of the containing
CS/κC/EEOS different
compositeconcentrations
films are of
shown EEOS
Table 1. Color parameters, opaqueness, and pictures of different in were
Table prepared.
1. The optical properties
EEOS concentration composite
of purple
the 1. tomato
CS/κC/EEOS anthocyanin
composite [31], chitosan/PVA/ZnO
films are shown in Table films containing
1. EEOS concentration purple potato or
films.
Table Color parameters, opaqueness, and pictures of different composite
rose anthocyanins [7], and cassava starch films containing lycium berry anthocyanins [27].
Table
films. 1. Color parameters, opaqueness, and pictures of different EEOS concentration composite
Film Sample a * b * L * Opaqueness
Table 1. Color parameters, opaqueness, and pictures of different EEOS concentration composite
films. s/% Picture
films. Table Color parameters,aopaqueness,
Film 1.Sample * and
b *pictures of different
L * EEOS concentration
Opaqueness s/% composite
Picturefilms.
Film Sample
CS/кC film a * b *
−1.22 ± 0.04 −0.18 ± 0.08 90.24 ± 0.33
d d L * a Opaqueness
1.06 ± 0.13 s/%
d Picture
Film Sample Film
a * Sample ba* * b* L* L * Opaqueness
Opaqueness s/% s/% Picture
Picture
CS/кC film −1.22 ± 0.04 d −0.18 ± 0.08 d 90.24 ± 0.33 a 1.06 ± 0.13 d
CS/кC film −1.22 ± 0.04 −0.18 ± 0.08 90.24 ± 0.33
d d a 1.06 ± 0.13 d
d 0.04d d −0.18 90.24 a ± 0.33 a
CS/кC film −1.22CS/кC
± 0.04 film −−1.22
0.18 ±±0.08 ± 0.08±d0.33
90.24 1.06 ± 0.13± d0.13 d
1.06
CS/кC/EEOS Ⅰ film 5.40 ± 0.25 c 10.62 ± 0.36 c 78.34 ± 0.39 b 3.55 ± 0.28 c
CS/кC/EEOS Ⅰ film 5.40 ± 0.25 c 10.62 ± 0.36 c 78.34 ± 0.39 b 3.55 ± 0.28 c
CS/кC/EEOS I film 5.40 ± 0.25 c Ⅰ film10.62
CS/кC/EEOS 0.25c c 10.62 78.34
5.40±±0.36 ± 0.36±c0.39
78.34b ± 0.39 3.55b 0.28± c0.28 c
3.55
±
CS/кC/EEOS Ⅰ film 5.40 ± 0.25 c 10.62 ± 0.36 c 78.34 ± 0.39 b 3.55 ± 0.28 c
CS/кC/EEOS Ⅱ film 12.68 ± 0.41 b 16.24 ± 0.54 b 66.40 ± 0.87 c 4.74 ± 0.07 b
CS/кC/EEOS II film 12.68 ± 0.41 Ⅱ film16.24
CS/кC/EEOS b 12.68±±0.54 b
0.41 16.24 66.40
b ± 0.54± 0.87
b c
66.40 ± 0.87 4.74 ±
c 0.07± b0.07 b
4.74
CS/кC/EEOS Ⅱ film 12.68 ± 0.41 16.24 ± 0.54 66.40 ± 0.87
b b c 4.74 ± 0.07 b
CS/кC/EEOS Ⅱ film 12.68 ± 0.41 b 16.24 ± 0.54 b 66.40 ± 0.87 c 4.74 ± 0.07 b
CS/кC/EEOS III film 17.26 ± 0.23 Ⅲ film18.06
CS/кC/EEOS a 17.26±±0.85 a
0.23 18.06 59.88
a ± 0.85± 0.91
a d
59.88 ± 0.91 5.56 ±
d 0.04± a0.04 a
5.56
CS/кC/EEOS Ⅲ film 17.26 ± 0.23 18.06 ± 0.85 59.88 ± 0.91
a a d 5.56 ± 0.04 a
CS/кC/EEOS
a–d
a–d Values Values Ⅲ film
arearegiven
given as 17.26
asmeans
means
± 0.23 a 18.06 ± 0.85 a 59.88 ± 0.91 d
± standard deviation.
± standard Different
deviation. letters in letters
Different
5.56
the sameinline ± 0.04
indicate
a
sameasignificant difference
(p < 0.05). Ⅲ film 17.26 ± 0.23 18.06 ± 0.85 59.88 ± 0.91
CS/кC/EEOS a a d 5.56the± 0.04 line indicate
significant
a–d Values difference
are given (pas <means
0.05). ± standard deviation. Different letters in the same line indicate
a–d Values are given as means ± standard deviation. Different letters in the same line indicate
significant difference (p < 0.05).
3.5.
ForAnalysis
a–d Values
significant of Mechanical
areCS/κC/EEOS
given
difference
the as <means
(p 0.05). ±Properties
films,standard deviation.
the a * value Different
increased fromletters
−1.22in± 0.04
the same line±indicate
to 17.26 0.23 (p
significantThe
difference (p < 0.05).
mechanical properties of composite films are expressed by tensile strength and
Forthe
< 0.05); thebCS/κC/EEOS films, from
* value increased the a −0.18
* value increased
± 0.08 from
to 18.06 −1.22(p±<0.04
± 0.85 toand
0.05) 17.26showed
± 0.23 (p
a
Polymers 2022, 14, x FOR PEER REVIEW Forthe
< 0.05); theincreasing
significant bCS/κC/EEOS
elongation * at
value films,
the break.
increased
trend the
Tensile
(p < from a −0.18
0.05).*Itvalue
strength increased
± (TS)
0.08 and
indicated to from −1.22(p
elongation
18.06
that the ±redness
0.85 at±value
<0.04
the to
break
0.05) 17.26
(EB)
and
and ± are
0.23
showed
yellowness (pa
11 ofrespec-
20
Forthe
<significant
0.05); the CS/κC/EEOS
bcomposite
* value films,
increased the a −0.18
* value increased from −1.22(p±<of
0.04 to and
17.26 ± 0.23 (paThe
valuetively
of theused to reflect
increasing trend the
films (p <from
mechanical ± 0.08
*resistance
0.05). ItLindicated
increased. to
decreased 18.06
and
that
from ±redness
0.85
theflexibility
90.24 0.05)
value
± 0.33 food
to and
59.88 ±showed
packaging.
yellowness
0.91 (p >
< 0.05); the
significant b * value
increasing increased
trend of(p from
< 0.05). −0.18 ±
ItLfilms 0.08
indicated to 18.06
thatfrom
the ± 0.85
redness (p < 0.05)
value and showed a
mechanical
value
0.05), of the
and the properties
composite
opacity films
of the the CS/κC/EEOS
increased.
composite composite
* decreased
increased films
90.24are shown
± 0.33
significantly, to and
59.88
indicatingyellowness
in Figure 4.(p
± 0.91
that the>
significant
value increasing trend (p < 0.05). It indicated that the redness value and yellowness
0.05), of
color of the
and the
the composite
opacity of
composite films
the increased.
films composite
gradually Lfilms
*deepened,
decreased from
increased 90.24
the ±brightness
significantly,
while 0.33indicating
to 59.88
and ± 0.91
that (p >
the
opacity
value and
0.05), of thethecomposite
opacity films
of the increased.
composite Lfilms
* decreased from
increased 90.24 ± 0.33indicating
significantly, to 59.88 ± 0.91
that (p >
the
color of the
gradually composite
decreased. Thefilmscolor gradually deepened,
of the composite filmswhile
graduallythe brightness
changed from andcolorless
opacity
0.05),
color and
of thethe opacity
composite of the
films composite
gradually films increased
deepened, significantly,
while indicating
theconcentration,
brightness and that the
opacity
gradually
and decreased.
transparent to redThe
and color
yellow.of the composite
With the filmsin
increase gradually
EEOS changed from thecolorless
color of
color of
gradually the composite
decreased. films gradually deepened, while the brightness and opacity
andfilms
the transparent
gradually redTheandcolor
to deepened, of the
yellow.
which composite
With
was films
the increase
consistent ingradually
with EEOS
the images changed
concentration, from
of the films. colorless
theThecolor of
color
gradually
and decreased.
transparent to redThe
andcoloryellow.of the composite
With filmsingradually
thebyincrease EEOS changed from
concentration, colorless
theThe
color of
the films
change ingradually
the CS/κC deepened,
film may which was consistent
be caused EEOS.with
Manythe images
studies ofhave
the films.
reported color
and
the transparent
films gradually to deepened,
red and yellow. which With the increase
was plant
consistent in EEOS
with the imagesconcentration,
ofhave
thefilms
films.theThe
color of
color
change
changes in
in the CS/κC
biopolymer film
films may be caused
containing by extracts,
EEOS. Many
such studies
as chitosan reported color
containing
the films
change ingradually deepened, which was consistent with the images of the films. The color
purple inthe
changestomato CS/κC
biopolymer film
anthocyaninfilms may bechitosan/PVA/ZnO
caused
containing
[31], by extracts,
plant EEOS. Many studies
suchcontaining
films as chitosanhave reported
films
purple color
containing
potato or
change
changes inin the CS/κC film
biopolymer films may be caused
containing by extracts,
plant EEOS. Many such studies
as havefilms
chitosan reported color
containing
purple
rose tomato anthocyanin
anthocyanins [7], and cassava [31], chitosan/PVA/ZnO
starch films containing films containing
lycium purple potato
berry anthocyanins or
[27].
changes
purple in biopolymer
tomato anthocyaninfilms [31],
containing plant extracts,films
chitosan/PVA/ZnO such containing
as chitosanpurplefilms containing
potato or
rose anthocyanins [7], and cassava starch films containing lycium berry anthocyanins [27].
purple tomato anthocyanin
roseAnalysis
anthocyanins [31], chitosan/PVA/ZnO
[7], andProperties
cassava starch films containing films containing
lycium purple potato
berry anthocyanins or
[27].
3.5. of Mechanical
rose anthocyanins [7], and cassava starch films containing lycium berry anthocyanins [27].
3.5. Analysis of Mechanical
The mechanical Properties
properties of composite films are expressed by tensile strength and
3.5. Analysis of Mechanical Properties
The
elongation mechanical
at the properties
break.
3.5. Analysis of Mechanical PropertiesTensile of strength
composite(TS) films
andareelongation
expressed by at tensile
the breakstrength
(EB) andare
The
elongation mechanical
at the properties
break. Tensile of composite
strength films
(TS) andare expressed
elongation
respectively used to reflect the mechanical resistance and flexibility of food packaging. by
at tensile
the strength
break (EB) and
are
The mechanical
elongation at the properties
break. Tensile of strength
composite(TS) films
andareelongation
expressed by at tensile
the strength
break (EB) and
are
respectively
The mechanical used to reflectofthe
properties themechanical
CS/κC/EEOS resistance
composite and flexibility
films are shown of food packaging.
in Figure 4.
elongation
respectively atused
the to
break.
reflectTensile strength resistance
(TS) and elongation at the break (EB) are
The mechanical properties ofthe
themechanical
CS/κC/EEOS composite andfilms
flexibility
are shown of food packaging.
in Figure 4.
respectively
The mechanical used to reflectofthe
properties themechanical
CS/κC/EEOS resistance
composite andfilms
flexibility
are shown of food packaging.
in Figure 4.
The mechanical properties of the CS/κC/EEOS composite films are shown in Figure 4.

Figure 4. Tensile strength and elongation at the break of the CS/κC, CS/кC/EEOS I, CS/кC/EEOS II,
Figure 4. Tensile strength and elongation at the break of the CS/κC, CS/кC/EEOS I, CS/кC/EEOS II,
a–d Values are given as means ± standard deviation. Different
and and CS/кC/EEOS
CS/кC/EEOS III composite
III composite films.films.
a–d Values are given as means ± standard deviation. Different
letters
letters in same
in the the same
line line indicate
indicate significant
significant difference
difference (p < (p < 0.05).
0.05).

When the content of EEOS was 0–5%, the tensile strength of the films decreased from
When the content of EEOS was 0–5%, the tensile strength of the films decreased from
9.07 to 4.27 MPa, and the elongation at the break increased from 22.37% to 34.87% with
9.07 to 4.27 MPa, and the elongation at the break increased from 22.37% to 34.87% with
the increase in the content of EEOS. The additive amount of EEOS affected the TS and EB
the increase in the content of EEOS. The additive amount of EEOS affected the TS and EB
(p < 0.05). With the rise in the EEOS addition level, TS showed a downward trend, which
(p < 0.05). With the rise in the EEOS addition level, TS showed a downward trend, which
may be due to the damage to the polymer network caused by excessive anthocyanins in
may be due to the damage to the polymer network caused by excessive anthocyanins in
the extract, resulting in a reduction in TS [26]. At the same time, all the films added with
EEOS showed greater EB than the CS/κC films. EB showed an opposite trend to TS. The
decrease in TS in concurrence with the increase in EB suggested increasing flexibility of
the films due to plasticization effects [32]. EEOS could change the mechanical properties
 Polymers 2022, 14, 2986 11 of 19

the extract, resulting in a reduction in TS [26]. At the same time, all the films added with
EEOS showed greater EB than the CS/κC films. EB showed an opposite trend to TS. The
decrease in TS in concurrence with the increase in EB suggested increasing flexibility of
the films due to plasticization effects [32]. EEOS could change the mechanical properties
of the films, which may be caused by the molecular interaction between anthocyanins in
the extract and the film-forming matrix [33]. Sun et al. found a similar decrease in TS and
an increase in EB in κC/HMx/Pmy membranes, which they attributed to the formation of
new hydrogen bonds between anthocyanins and polymers. The results coincided with the
FITR and XRD analyses [12].

3.6. WVP
Due to the role of water in metamorphic reactions, WVP is an important index to
evaluate the physical properties of films and a vital barrier parameter to prevent water
transfer between food and the environment, reflecting the water permeability potential and
Polymers 2022, 14, x FOR PEER REVIEW barrier performance of food-packaging films. In general, low water vapor permeability
12 of 20 is
preferred because of its high sealing effect and its conduciveness to the long-term storage
of food materials [26]. The WVP values of the CS/κC/EEOS composite films are shown in
Figure 5.

Figure 5. WVP of CS/κC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III compositea–dfilms.
Figure
a–d 5. WVP of CS/κC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III composite films.
Values are given as means ± standard deviation. Different letters in the same line indicate
Values are given as means ± standard deviation. Different letters in the same line indicate significant
significant
difference difference (p < 0.05).
(p < 0.05).

The WVP value of the CS/κC film was 1.08 g·mm/m2 ·d·kPa, and the WVP values
The WVP value of the CS/κC film was 1.08 g·mm/m2·d·kPa, and the WVP values of
of the composite films increased with the increase in EEOS from 1% to 5%. It can be seen
the composite films increased with the increase in EEOS from 1% to 5%. It can be seen that
that the WVP values of the films did not change significantly with the addition of a small
the WVP values of the films did not change significantly with the addition of a small
amount of EEOS (p > 0.05). Still, when the EEOS addition level reached 5%, the WVP values
amount of EEOS (p > 0.05). Still, when the EEOS addition level reached 5%, the WVP
of the composite films increased significantly (p < 0.05) and reached 1.71 g·mm/m2 ·d·kPa.
values of the composite films increased significantly (p < 0.05) and reached 1.71
This indicated that the addition of EEOS affected the physical properties of the CS/κC films,
g·mm/m2·d·kPa. This indicated that the addition of EEOS affected the physical properties
and a higher content of EEOS (5%) led to a significant increase in WVP value (p < 0.05). This
of the CS/κC films, and a higher content of EEOS (5%) led to a significant increase in WVP
may be because the excessive anthocyanins in EEOS damage the dense molecular structures
value (p <films,
of the 0.05).leading
This may be because
to uneven the excessive
dispersion of the filmanthocyanins in EEOS
matrix and a small damage
number the
of insoluble
dense molecular structures of the films, leading to uneven dispersion of the film matrix
particles that make the film surface porous [26]. Moreover, plasticization effects increase
and a small number of insoluble particles that make the film surface porous [26].
Moreover, plasticization effects increase the molecular mobility and diffusion rates of
water vapor through film matrices, which increases WVP [34]. Similar studies have also
confirmed that the addition of plant extracts can improve the WVP values of starch-based
films [27,34,35]. Tavares et al.’s experiments showed that the addition of a CMC polymer
 Polymers 2022, 14, 2986 12 of 19

the molecular mobility and diffusion rates of water vapor through film matrices, which
increases WVP [34]. Similar studies have also confirmed that the addition of plant extracts
can improve the WVP values of starch-based films [27,34,35]. Tavares et al.’s experiments
showed that the addition of a CMC polymer to a maize starch film reduced the WVP values.
Their results indicated that the formation of CMC polymeric blends with starch improved
water resistance to some extent [35]. This was similar to a study on cassava starch films
mixed with Chinese bayberry anthocyanins [27]. Excessive extract formed aggregates and
produced more free volumes in the film network to facilitate moisture transfer.

3.7. DSC
Polymers 2022, 14, x FOR PEER REVIEW 13 of 20
DSC is a technique to study the interactions between polymer molecules. Tg appears
as an endothermic shift. The peak should be a first-order phase transition. It can be
used to analyze the compatibility and thermal stability of blends. It reflects the molecular
calorimetry analysis
movements was performed
of various types ofon the CS/κC/EEOS
polymers and has composite films. A DSC
a great relationship diagram
with the kinds
of the
ofcomposite
materials films is shown in
and structural Figure 6. of composite films [36]. A differential scanning
properties
calorimetry analysis was performed on the CS/κC/EEOS composite films. A DSC diagram
of the composite films is shown in Figure 6.

FigureFigure 6. Thermodynamic
6. Thermodynamic propertiesproperties of the
of the CS/κC, CS/κC, CS/кC/EEOS
CS/кC/EEOS I, CS/кC/EEOSI,II,CS/кC/EEOS II, and
and CS/кC/EEOS
CS/кC/EEOS
III composite films. III composite films.

The The endothermic

endothermic peak peak
showed showed that
that the Tgthe Tg of
of the the CS/κC
CS/κC film, thefilm, the CS/κC/EEOS
CS/κC/EEOS Ⅰ film, I
film, the CS/κC/EEOS II film, and the CS/κC/EEOS III film were 133.77 ◦ C, 125.12 ◦ C,
the CS/κC/EEOS Ⅱ film, and the CS/κC/EEOS Ⅲ film were 133.77 °C, 125.12 °C, 124.18 °C,
124.18 ◦ C, and 123.20 ◦ C, respectively. Both the CS/κC film and the composite films had
and 123.20 °C, respectively. Both the CS/κC film and the composite films had peaks
peaks between 20 ◦ C and 250 ◦ C, which indicated that the compounds in the composite
between 20 °C and 250 °C, which indicated that the compounds in the composite films
films had good compatibility. The glass-transition temperatures of the composite films
had good compatibility. The glass-transition temperatures of the composite films added
added with EEOS were lower than that of CS/κC, and the glass-transition temperatures of
with EEOS were lower than that of CS/κC, and the glass-transition temperatures of the
the composite films decreased with the increase in EEOS. Similar results were observed in
composite films decreased with the increase in EEOS. Similar results were observed in the
the study of Wang et al. [37]. The decrease in the glass-transition temperature may be due
study of Wang et al. [37]. The decrease in the glass-transition temperature may be due to
to the damage of starch chain interaction caused by extract of onion skin and the increase
the damage of starch chain interaction caused by extract of onion skin and the increase in
in chain flexibility.
chain flexibility.
3.8. TPC
3.8. TPC
It is well-explained that naturally extracted polyphenolic compounds are excellent
It is well-explained
antioxidants. thatthere
Therefore, naturally extracted
is a close polyphenolic
relationship betweencompounds
the phenolicare excellent
contents and the
antioxidants. Therefore, there is a close relationship between the phenolic contents and
the antioxidant activity [30]. The TPC assay determines the amounts of total phenolic
compounds in the samples, which are responsible for the antioxidant activity [38].
Therefore, the polyphenol content can be used as an essential indicator for evaluating the
antioxidant capacity. Figure 7 shows the total phenolic contents of the CS/κC/EEOS
 Polymers 2022, 14, 2986 13 of 19

antioxidant activity [30]. The TPC assay determines the amounts of total phenolic com-
pounds in the samples, which are responsible for the antioxidant activity [38]. Therefore,
Polymers 2022, 14, x FOR PEER REVIEW 14 of 20
the polyphenol content can be used as an essential indicator for evaluating the antioxidant
capacity. Figure 7 shows the total phenolic contents of the CS/κC/EEOS composite films
with different amounts of EEOS addition.

Figure 7. Total phenol contents of the CS/κC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS
Figure 7. Total phenol contents of the CS/κC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III
III composite films. a–d Values are given as means ± standard deviation. Different letters in the same
composite films. a–d Values are given as means ± standard deviation. Different letters in the same
line indicate significant difference (p < 0.05).
line indicate significant difference (p < 0.05).
The CS/κC composite film itself had a very low TPC content. When EEOS was
The CS/κC composite film itself had a very low TPC content. When EEOS was added
added to the CS/κC composite films, the total phenolic content of the composite films
to the CS/κC composite films, the total phenolic content of the composite films increased
increased significantly with EEOS concentration. The total phenol content increased from
significantly with EEOS concentration. The total phenol content increased from 1.61
1.61 GAEmg/DWg to 6.73 GAEmg/DWg with the rise in EEOS from 1% to 5%. The TPC
GAEmg/DWg to 6.73 GAEmg/DWg with the rise in EEOS from 1% to 5%. The TPC (6.73
(6.73 GAEmg/DWg) content of the CS/κC/EEOS III composite film with 5% EEOS content
GAEmg/DWg) content
was the highest. of the
Purple onion skin is richⅢincomposite
CS/κC/EEOS anthocyaninsfilm and
withother
5% EEOS content
phenolic was
compounds,
the highest. Purple onion skin is rich in anthocyanins and other phenolic compounds,
and the content of polyphenols is closely related to antioxidant activity, which significantly and
the increased
content ofthepolyphenols is closely
total phenolic contents related
of theto antioxidant
composite activity,
films, which significantly
thus improving the biological
increased the total phenolic contents of the composite films, thus improving
activity of the composite films. It can also be interpreted that anthocyanin contains the biological
phenolic
activity of the composite
compounds films.hydrogen
that can provide It can also be interpreted
atoms that the
[39]. The higher anthocyanin contains
concentration of EEOS,
phenolic compounds
the more that can
anthocyanins provide
it has, hydrogen
the more phenolicatoms [39]. Theithigher
compounds has, andthethe
concentration
more potent its
of EEOS, the more anthocyanins
antioxidant capacity. it has, the more phenolic compounds it has, and the more
potent its antioxidant capacity.
3.9. Analysis of Antioxidant Properties In Vitro
3.9. Analysis of Antioxidant
Antioxidant Properties
capacity In Vitrofor active food packaging because free radicals
is essential
Antioxidant
can capacity
cause food is essential
spoilage for active
and nutrient foodThe
loss [33]. packaging
in vitrobecause free radicals
antioxidant capacitycan
of the
cause food spoilage
CS/κC/EEOS and nutrient
composite films isloss
shown[33]. The in8.vitro antioxidant capacity of the
in Figure
CS/κC/EEOS composite films is shown in Figure 8.
Polymers 2022, 14, x FOR PEER REVIEW 15 of 20
Polymers 2022, 14, 2986 14 of 19

Figure 8. Scavenging activities of the CS/κC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III
Figure 8. Scavenging activities of the CS/κC, CS/кC/EEOS I, CS/кC/EEOS II, and CS/кC/EEOS III
composite films on DPPH and ABTS radicals. a–d Values are given as means ± standard deviation.
composite films on DPPH and ABTS radicals. a–d Values are given as means ± standard deviation.
Different letters in the same line indicate significant difference (p < 0.05).
Different letters in the same line indicate significant difference (p < 0.05).
The single CS/κC film without EEOS had weak scavenging ability for DPPH and ABTS
The single CS/κC
free radicals. film
With the without
increase in EEOS
the EEOS hadcontent,
weak scavenging ability for DPPHability
the free-radical-scavenging and of
ABTSthefree radicals.films
composite With thegradually
also increase improved.
in the EEOS Whencontent, the free-radical-scavenging
the EEOS additive level was 5%, the
ability of the composite films also gradually improved.
DPPH free-radical-scavenging rate reached 45.14%. The ABTS When the EEOS additive level was
radical-scavenging ability
5%, was
the DPPH free-radical-scavenging rate reached 45.14%. The ABTS
up to 69.50%. The content of EEOS in the composite films was directly proportional radical-scavenging
ability wasfree-radical-scavenging
to the up to 69.50%. The content activity, of which
EEOS indicated
in the composite films was directly
that the anthocyanin content in
proportional to the free-radical-scavenging activity, which indicated
EEOS had a significant influence on the antioxidant capacity of the composite that the anthocyanin
films. The
content in EEOS had a significant
free-radical-scavenging abilityinfluence on the antioxidant
of the CS/κC/EEOS capacity
composite of the
films wascomposite
significantly
films. The free-radical-scavenging
enhanced with EEOS, which was mainly ability attributed
of the CS/κC/EEOS compositesuch
to active components, films was
as phenolic
significantly enhanced with EEOS, which was mainly attributed
compounds and secondary metabolites, that can inhibit oxidation in plants [40]. The to active components,
suchincrease
as phenolic compounds and secondary
in free-radical-scavenging activity was metabolites,
also foundthat can
in the inhibit
purple oxidation
cabbage in
component
plants [40]. The increase in free-radical-scavenging activity was
of anthocyanin-and-biopolymer-based composite films [41]. The polyphenols in EEOS also found in the purple
cabbage component
are mainly of anthocyanin-and-biopolymer-based
anthocyanin. The anthocyanins that are rich incomposite EEOS could films [41]. phenolic
provide The
polyphenols
hydroxyl in to EEOS
capturearefree
mainly anthocyanin.
radicals, which made The upanthocyanins that areinrich
for the deficiency thein EEOS
antioxidant
could provide
activity of thephenolic hydroxyl to
CS/κC composite filmscapture free radicals,
and improved which made
the antioxidant up of
activity forthethe
films.
deficiency in thesuggested
These results antioxidant thatactivity of theofCS/κC
the presence compositeinfilms
the polyphenols EEOSand improved
and the presence the
of the
antioxidant
phenolic activity
compounds of the films.
in plant Thesecontributed
extracts results suggested
to enhancing thatthe
thefree-radical-scavenging
presence of the
polyphenols
potential,in EEOSenhancing
thereby and the presence of the activity.
the antioxidant phenolicThe compounds
CS/κC/EEOS in plant extractsfilms
composite
contributed
containing to EEOS
enhancing
had athe free-radical-scavenging
high antioxidant capacity and potential,
could thereby
prevent enhancing
lipid oxidationthe in
antioxidant activity.the
food. Therefore, The CS/κC/EEOS
composite films madecomposite
in this films
study containing
can be usedEEOS had apackaging
as an active high
materialcapacity
antioxidant to delay and
lipidcould
oxidation
preventin foods
lipid with high fat
oxidation content
in food. [14].
Therefore, the composite
films made in this study can be used as an active packaging material to delay lipid
3.10. Analysis
oxidation in foodsofwithOxidation
high fatResistance
content of the Film
[14].
Good oxidation resistance is significant for active food-packaging film. Therefore, we
3.10.chose lardofasOxidation
Analysis the experimental
Resistancematerial to evaluate the antioxidant capacity of the composite
of the Film
films. The POV values representing
Good oxidation resistance is significant for the oil oxidation performances
active food-packaging of the
film. CS/κC/EEOS
Therefore, we
chose lard as the experimental material to evaluate the antioxidant capacity of the
Polymers 2022, 14, x FOR PEER REVIEW 16 of 20

Polymers 2022, 14, 2986 15 of 19

composite films. The POV values representing the oil oxidation performances of the
CS/κC/EEOS composite films are shown in Figure 9, and the POV values are proportional
to composite films
the oxidation are shown in Figure 9, and the POV values are proportional to the oxidation
degree.
degree.

Figure 9. POV values of lard coated with CS/κC/EEOS composite films with different EEOS
Figure 9. POV values of lard coated with CS/κC/EEOS composite films with different EEOS additive
additive levels.
levels.
The oxidation degree of unwrapped lard increased from 0.24 to 2.61 g/100 g within
7 days.oxidation
The The POVdegree value of oflard
unwrapped
decreased lard increased
obviously from
after 0.24parceled
being to 2.61 g/100
by theg within
composite 7
days. The POV value of lard decreased obviously after being
films, which indicated that the composite films had a blocking effect on oxygen, reduced parceled by the composite
films, which indicated
the contact between the thatsample
the composite
and oxygen, filmsand had a blocking
inhibited the effect
oxidationon oxygen,
degree of reduced
lard. The
thecomposite
contact between the sample and oxygen, and inhibited
films without EEOS had poor inhibition effects on lard oxidation. On the the oxidation degree of 7th
lard. day,
The composite films without EEOS had poor inhibition effects
the POV value of lard reached 1.01 g/100 g, which was higher than that of the composite on lard oxidation. On the
7thfilms
day,withthe EEOS,
POV value of lard
indicating thatreached 1.01 g/100
the addition of EEOSg, which
improved was thehigher than
ability thatcomposite
of the of the
composite films with
films to inhibit EEOS, indicating
oil oxidation and played that athe addition
certain of EEOSrole.
synergistic improved the ability
The addition of
of EEOS
thesignificantly
composite films to inhibit oil oxidation and played a certain
improved the antioxidant activity of the CS/κC films, which was related to synergistic role. The
addition
the strong of EEOS significantly improved
free-radical-scavenging ability ofthetheantioxidant
EEOS, which activity
was rich of in
theanthocyanins
CS/κC films, [38].
which
Gao et al. [22] also reached a similar conclusion that the POV values of lardwhich
was related to the strong free-radical-scavenging ability of the EEOS, wrapped was in
rich in anthocyanins
CS/CA/EBSC [38]. Gao
composite films et were
al. [22] also reached
significantly a similar
higher conclusion
than those that the
in CS/CA POV
films. With
values of lard wrapped
the increase in the EEOS in CS/CA/EBSC
additive level,compositethe EEOS filmscompositewere films
significantly higher
had a better than
inhibiting
those
effectin on
CS/CA
lard films.
oxidation,Withbutthethe increase
5% EEOS in the EEOS additive
composite films had level, the EEOS
a poor composite
inhibiting effect on
films
lardhad a better This
oxidation. inhibiting
showseffect that itonwaslardnot oxidation,
the higher butconcentration
the 5% EEOSof composite
EEOS, thefilms better
had a poor inhibiting effect on lard oxidation. This shows
the antioxidant effect of the composite film. Excessive EEOS affected the structure of the that it was not the higher
concentration
films, increased of EEOS, the better
the oxygen the antioxidant
penetration of the films,effect
and ofled
thetocomposite film. Excessive
lard oxidation. This may be
EEOS affected
because, withthe thestructure
increase in of the
theamount
films, increased
of EEOS, the microstructures
oxygen penetration of the films, of
and compactness
andtheledcomposite films wereThis
to lard oxidation. affected,
may be which
because,weakened
with thetheincrease
ability ofinthe thefilms to block
amount oxygen.
of EEOS,
theHowever, the POVand
microstructures value of the lard wrapped
compactness with 3% EEOS
of the composite films waswere0.34 g/100 which
affected, g on the
7th day, the
weakened indicating
ability of that the
the composite
films to blockfilms at this
oxygen. concentration
However, the POV hadvalue
the best inhibition
of the lard
effect on lard oxidation. Similarly, Nilsuwan et al.
wrapped with 3% EEOS was 0.34 g/100 g on the 7th day, indicating that the composite found that, when epigallocatechin
gallate
films (EGCG)
at this was added
concentration had to athefish gelatin
best membrane,
inhibition effect iton could
lardmigrate
oxidation.to high-fat
Similarly, food
mimics [42].
Nilsuwan et al.The foundresults
that,showed that composite films
when epigallocatechin gallate of biodegradable
(EGCG) was added films andto anatural
fish
activemembrane,
gelatin substancesithave could potential
migrateapplication
to high-fat value in food[42].
food mimics packaging.
The results showed that
Polymers 2022, 14, 2986 16 of 19

3.11. Bacteriostatic Analysis

The size of the bacteriostatic rings of Staphylococcus aureus (Gram-positive) and Es-
cherichia coli (Gram-negative) can be used to evaluate the bacteriostatic activity of composite
film samples. The bacteriostatic ring diameters of the CS/κC/EEOS composite films against
S. aureus and E. coli are shown in Table 2.

Table 2. Diameter of bacteriostasis circles of CS/κC and CS/κC/EEOS composite films against
Escherichia coli and Staphylococcus aureus.

Diameter of the Bacteriostatic Circle (mm)

Extract Concentration
Escherichia coli Staphylococcus aureus
CS/кC film 7.06 ± 0.03 d 7.02 ± 0.05 d
CS/кC/EEOS I film 11.20 ± 0.14 c 11.48 ± 0.25 c
CS/кC/EEOS II film 12.63 ± 0.36 b 14.52 ± 0.22 b
CS/кC/EEOS III film 14.12 ± 0.32 a 15.40 ± 0.38 a
a–d Values are given as means ± standard deviation. Different lowercase letters in the same column indicate
significant difference (p < 0.05).

The inhibition effect of the CS/κC composite films on S. aureus and E. coli was not
pronounced. When EEOS was added to the composite films, the antibacterial effect of the
films was improved with the increase in the amount of EEOS. The diameter of the antibac-
terial ring was gradually increased with the rise in the amount of EEOS. Koosha et al. [43]
added black carrot anthocyanin to chitosan/PVA films and found that the antibacterial
performance of the chitosan/PVA composite films increased with the increase in black
carrot anthocyanin content. The antibacterial ring diameter of the composite films against
S. aureus was more extensive than that of E. coli, which indicated that the antibacterial
effect of the composite films against Gram-positive bacteria was better than that against
Gram-negative bacteria. Previous studies have reported similar results, suggesting that
plant extracts are more effective in killing S. aureus than E. coli [7]. The flavonoid com-
pounds contained in EEOS can reduce the fluidity of the cell membrane or perforate the
membrane through the hydrogen peroxide pathway, thus damaging the cell membranes of
microorganisms, leading to the secretion disorder of microbial cells and, thus, inhibiting
the growth of microorganisms. Previously, starch-based films with sappan and cinnamon
plant extracts showed antimicrobial efficiency that effectively extended the shelf-life of
packaged meat [44]. A study showed that the phenolic compounds in EEOS can not only
increase the permeability of the cell membrane but can also interfere with the synthesis of
microbial genetic material, thus inhibiting the growth of microorganisms and improving
the bacteriostatic performances of CS/κC composite films [45]. Liu et al. also reached a
similar conclusion when studying the inhibitory effect of CS/polyvinyl alcohol/graphene
oxide nanofibrous films loaded with allicin on three foodborne pathogens [46].

4. Conclusions
In this study, CS/κC/EEOS composite films were prepared by blending 0%~5%
ethanol extract of onion skin with corn starch/κ-carrageenan films, and the effects of EEOS
concentration on the physicochemical properties and biological activities of the composite
films were discussed. FTIR, XRD, and SEM analyses showed that the addition of EEOS
could inhibit the recrystallization of the starch to a certain extent. The microstructures of
the composite films were affected, and the surfaces and cross-sections of the films became
rough. When the EEOS concentration was 5%, the film structure was coarsest. The addition
of EEOS reduced the brightness, increased the opacity and deepened the film color. The
mechanical properties of the film were affected: the tensile strength decreased and the
elongation at the break increased; the WVP value increased; and DSC showed that the
glass-transition temperatures of the composite films were decreased by adding EEOS. In
terms of biological activity, the total phenolic content, as well as the DPPH and ABTS
free-radical-scavenging rates, increased significantly with EEOS concentration. When the
Polymers 2022, 14, 2986 17 of 19

EEOS concentration was 5%, the free-radical-scavenging ability was the highest. The results
showed that the composite films with an EEOS concentration of 3% had the best antioxidant
effects, and the POV value of lard wrapped with EEOS was the lowest on the 7th day. The
inhibitory effect of the films on S. aureus was higher than that of E. coli, and the diameter
of the inhibitory ring increased with the increase in EEOS concentration. From what was
discussed above, the optimal addition amount of EEOS was 3%, and that film had the best
all-around performance. The prepared composite film has potential application value in
food packaging. The results can provide a reference for the production and application of
CS/κC/EEOS composite films.

Author Contributions: Conceptualization, C.W.; writing—original draft preparation, Y.L.; writing—

review and editing, C.W.; methodology, Z.L.; data curation, X.A.; software, Z.G.; resources, S.T. All
authors have read and agreed to the published version of the manuscript.
Funding: The work was financially supported by Qiqihar University and the Foundation for the
Characteristic Discipline of Processing Technology of Plant Foods (No. YSTSXK201812).
Institutional Review Board Statement: Studies not involving humans or animals.
Informed Consent Statement: Studies not involving humans.
Data Availability Statement: The datasets generated and analyzed during the current study are
available from the corresponding author on reasonable request.
Acknowledgments: The authors appreciate the support from the Foundation for the Characteristic
Discipline of Processing Technology of Plant Foods and the Qiqihar University Testing Center.
Conflicts of Interest: The authors declare no conflict of interest.

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