Environmental Technology & Innovation 25 (2022) 102180

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Environmental Technology & Innovation

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

Intensification of yam-starch based biodegradable bioplastic

film with bentonite for food packaging application
∗
Lopamudra Behera a ,1 , Monalisha Mohanta b ,1 , Arunachalam Thirugnanam b ,
a
Department of Biotechnology, Ravenshaw University, Cuttack, 753003, India
b
Department of Biotechnology and Medical Engineering, National Institute of Technology Rourkela, Odisha, 769008, India

article info a b s t r a c t

Article history: Bioplastics are gaining more attention over synthetic plastics owing to their non-
Received 15 September 2021 hazardous properties. Earlier studies had reported the biodegradation and solubility
Received in revised form 15 November 2021 properties of yam starch-based bioplastic; however, its mechanical properties with
Accepted 5 December 2021
reinforcement were not studied. The present work focuses on the development of
Available online 10 December 2021
yam (Dioscorea) starch-based bioplastic film with glycerol as a plasticizer. Different
Keywords: concentrations of bentonite (0.5, 1, 1.5 %w/w) was reinforced and the biofilms were
Bioplastics prepared by solvent casting method. The developed bioplastic films were systematically
Yam-starch characterized using the scanning electron microscope (SEM), Fourier transforms infrared
Bentonite spectroscopy (FTIR), contact angle, tensile testing, soil and water degradation, water
Soil biodegradation
vapor transmission rate (WVTR), swelling, solubility, and chemical testing. The SEM
Tensile strength
micrographs show the bentonite particles were homogeneously distributed in the
starch matrix. The hydrophilicity of the bioplastic was increased with the addition of
bentonite. The FTIR spectra revealed that the addition of bentonite in the bioplastic
film strengthened the O–H bond and Si–O–Si bond, which improved the mechanical
strength. It was observed that 1.5% bentonite concentration exhibited maximum soil
degradation rate and lowest water molecule absorption. This is due to the intercalated
silicate layer of bentonite in the bioplastic films. Also, the bioplastic films with bentonite
were more impermeable in salt and acidic medium but remained ineffective in the
basic medium. The study revealed that bentonite concentration has effectively improved
the yam starch-based bioplastic property, making it a potential bioplastic to replace
synthetic plastic in food packaging applications.
© 2021 Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND
license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction

Plastic materials are the derived form of thermosetting and thermoplastic polymers. Thermosetting polymers get
solidified or fixed by applying heat. On the other hand, thermoplastic is a plastic polymer material that becomes flexible at
a specific raised temperature and sets after cooling. Thermoplastic constitutes a significant portion of plastic in food pack-
aging applications. It can be quickly framed economically into any expected shape to satisfy food packaging requirements.
It is particularly friendly to reusing and following the waste-to-energy process. The mainly used thermoplastic polymers
in food packaging are polyolefin (low thickness or high thickness polyethylene, polypropylene, and so on), polystyrene,

∗ Correspondence to: Department of Biotechnology & Medical Engineering National Institute of Technology Rourkela, Room No. 206, Odisha, PIN
– 769008, India.
E-mail address: thirugnanam.a@nitrkl.ac.in (A. Thirugnanam).
1 Authors have equal contribution.

https://doi.org/10.1016/j.eti.2021.102180
2352-1864/© 2021 Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

polyester, and nylon polycarbonate, and vinyl polymers. These polymers provide excellent mechanical reliability and
incredible strength, the prime requirement for food packaging application (Grigore, 2017; Marsh and Bugusu, 2007).
Polyethylenes are polymers mainly comprised of carbon (C) and hydrogen (H2 ), extraordinarily impervious to natural
degradation. Their gatherings can be dangerous and cause some natural aggravation. The traditional techniques utilized
for polymer debasement include landfill, burning, and substance treatment, making a destructive impact on the climate.
Also, these plastics provide a significant threat to human beings due to their non-biodegradable nature (Bhuyar et al.,
2018). It was reported that annual plastic production has gradually increased from 1950 (35 million metric tons) to 2018
(exceed 350 million metric tons) throughout the world, and 93% of them have usually been discarded in-ground (landfills
site) and seas (Dos Santos et al., 2019; Peixoto et al., 2019). The plastics disposed of in landfills emit about 253 g carbon
dioxide (CO2 ) per kg, while ignition discharges 673–4605 g kg−1 of CO2 . This incineration practice leads to enormous
greenhouse gas emissions, which contribute to global warming. It is assessed that polythene would debase only 0.5% even
after 100 years. Its degradation essentially relies upon temperature, light, oxygen (O2 ), and dampness accessibility (Bhuyar
et al., 2019). Also, partial combustion of plastics releases carbon monoxide (CO) and other hazardous gas directly into the
environment, thereby polluting the environment (Eriksson and Finnveden, 2009). Plastics contain diethylhexyl phthalate
(DEHP) and bisphenol-A, along with phthalates which are toxic and lead to cancer. It also affects the human reproductive
system and creates genital defects in a human being (Talsness et al., 2009). The potential ways to deal with decrease
the plastic waste are to foster biodegradable item plastics from a non-renewable energy source or reengineering of total
carbon spine product polymers and to distinguish possible micro green algae with their toxins to enable convention to
adequately biodegrade the plastics (Bhuyar et al., 2021; Jadhav et al., 2021). Hence, plant-based extracts have been utilized
for making eco-friendly methods for numerous applications (Sundararaju et al., 2020; Khazaai et al., 2021).
Therefore, there is a huge demand for plastics manufacturing from renewable resources, commonly called ‘‘Bioplastic’’,
to replace synthetic polymer-based plastics, especially in food packaging industries. These plastics are obtained from
natural resources and are biodegradable. Sources such as potatoes, rice, maize, etc., are used to develop bioplastics because
they are low-cost, abundant, readily available, and form incessant matrixes (Acquavia et al., 2021). These sources contain
starch rich in glucan, primarily involved in two major fundamental components as amylose and amylopectin. The starch
chain has many hydroxyl groups (O–H) in which two subordinate O–H at C-2 and C-3 of all glucose excess and one prime
O–H at C-6 are not interconnected. This phenomenon leads the starch to be hydrophilic. Amylose contains linear molecules
of glucose elements connected with the α (1–4) bond. Amylopectin contains bifurcated molecules with α (1–6) bonds
which linked branch units with the linear chain of α (1–4) glucose. Starch behaves as a semi-crystalline biopolymer that
assists as a reservoir of carbohydrates in many floras containing seeds, tubers, and cereals. Also, starch possesses excellent
thermal conductivity and biodegradable properties (Navarro et al., 2019).
Yam (Dioscorea) has 600 species, and among them, ten species are stapled yam. The amylase content in yam is about
30% which has great potential to form films. Yam starch films contain a homogeneous matrix which provides a steady
structure at ambient conditions. Also, starch films possess remarkable O2 hindrance properties because of their firmly
pressed systematic H2 bonded grouped and low dissolvability (Dos Santos Caetano et al., 2018). But the disadvantages of
starch films are their brittleness behavior (Vu et al., 2016). So, to overcome this innate of starch films, the integration
of a plasticizer is necessary. A plasticizer (glycerol) is added for making smooth films, thereby enhancing the films’
flexibility. The effect of plasticizers on the mechanical properties of abundant starch films illustrates a significant water
vapor transmission rate and higher solubility (Souza et al., 2012). Fillers are necessary to introduce plasticizers to lower
the shrinkage, cost, and better functionality of plastic. The filler is essential in product development for a good variety
of bioplastics. Usually, starch films are fragile and difficult to handle; many researchers have shown that filler is used as
reinforcement in thermoplastic materials to improve the mechanical behavior of starch-based bioplastics films. Generally,
filler particle size affects the tensile strength, elasticity modulus, and elongation capacity of bioplastic film (Bodiralu
et al., 2013). Bentonite contains a good molecule size that prompts significant interaction with starch composite lattices,
confirming good cooperation with different particles in composite film networks (Alekseeva et al., 2017). Bentonite
comprises significantly Montmorillonite (Al2 O3 ·4(SiO2 )·H2 O), which belongs to the 2:1 phyllosilicates group of clay with
tetrahedral and octahedral sheets facilitated together, shaping layers. The above configuration is commonly called as
Hoffmann structure (Hofmann and Endell, 1933).
Research interest on the yam starch is gained from an industrial perspective because it is considered a latent starch
source that can form bioplastic films. Mali et al. (2005) have evaluated the mechanical property of yam starch with
glycerol as a plasticizer. They observed with an increase in the plasticizer concentration, the puncture deformation, and
strain increased whereas the tensile stress and Young’s modulus decreased. Omotoso et al. (2015) have also evaluated the
effect of plasticizer (glycerol) on the mechanical properties of starch-based film. The study reported that 59% of glycerol
content increased the tensile strength up to 5.74 MPa. Similarly, Rugchati and Thanacharoenchanaphas (2015) evaluated
the puncture properties of yam starch-based biofilm with glycerol and surfactants such as tween 20 and span80. Another
study reported that with the addition of chitosan and clove oil in yam starch, its antimicrobial and antibacterial properties
enhanced but did not affect their mechanical properties (Ulyarti et al., 2021). Agarwal, 2021 reviewed that the fillers used
with starch can increase tensile strength. However, there are no studies reported on the mechanical properties of yam
starch with bentonite reinforced filler.
In this study, a novel bioplastic film was developed using yam starch and bentonite as reinforcement filler. The effect
of filler content was characterized physiochemically, mechanically, and biologically. Further, biodegradability tests were
performed in-vitro to produce good biodegradable bioplastic film with good mechanical properties for food packaging,
which can potentially replace synthetic plastic packaging systems.
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

Fig. 1. Schematic representation of the development of Yam starch bioplastic film using solvent casting method.

2. Materials and methods

2.1. Materials

Fresh tubers of yam were obtained from a local market of the Sundergarh district of Odisha (India). Glycerol (98%
purity) was purchased from Merck private limited, India (DH8F681741). Purified bentonite powder (RM785-500G), sodium
hydroxide (NaOH) [RM467-500G], and sodium chloride (NaCl) were procured from HIMEDIA laboratories private limited.

2.2. Isolation of starch

Starch was isolated from fresh root tubers of yam about 3 g using a wet milling process (Sukhija et al., 2015). The root
tubers of yam were washed to clean off the adhering soil and chopped into small pieces. The pieces were homogenized
in water using a domestic blender. The slurry was filtered through an ultra-fine muslin cloth. The starch was allowed to
settle down, and the supernatant was poured out. The mixture was then filtered to get a simple starch cake. Then the wet
starch was finely spread onto dry trays and dried in a hot air oven at 50 ◦ C for 48 h. The dried starch was then crushed to
fine particle size by a conventional blender, and blended powder was passed through a sieve size of 300 µm and 150 µm
(standard test sieve as per ISS: 46-1962) according to the particle dimension. The obtained yam powder was stored in an
airtight jar for characterization.

2.3. Preparation of starch-based bioplastic films

Bioplastic films were prepared by using 3% (w/w) starch powder, glycerol (plasticizer) of about 1.5% (w/w) and with
three different proportion of bentonite (filler) powder (0.5%, 1%, and 1.5% w/w) in 100 mL of distilled water (DW) (Luckham
and Rossi, 1999; Kar et al., 2016; Shanmathy et al., 2021). Yam starch and plasticizer were directly mixed with distilled
water to make batches with a magnetic stirrer at room temperature for 15 min. The bentonite powder was then mixed
in the normalized solution at 400 rpm, and later the mixture was heated at 85 ◦ C until gelation occurred. Gelatinized
suspension starch was immediately poured on a circular Petri plate and allowed to dry in a ventilated hot air oven at 40
◦
C for 24 h. This process was repeated for all different proportions of bentonite powder, and all tests were performed in
triplicates. The procedure of the experimental work is schematically represented in Fig. 1, and the coding for the developed
yam starch-based biofilms is given in Table 1.
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

Table 1
Coding of the developed yam-starch based biofilm.
Chemical composition Filler (%w/w) Coding
0 B0
0.5 B1
3% (w/w) starch powder +1.5% (w/w) glycerol, plasticizer
1 B2
1.5 B3

2.4. Physicochemical characterization

2.4.1. Scanning electron microscope (SEM)

Isolated yam starch powder and the starch-based bioplastic film surfaces were studied using ESEM (environmental
scanning electron microscope) (FEI Quanta FEG 250, Netherlands) using 15 kV at 2000X magnification. The samples were
cut into 1 × 1 cm and were entirely dried in a hot air oven at 40 ◦ C for 5 h prior to ESEM analysis. The particle size of
yam starch powder was calculated using Image-J software (King et al., 2021).

2.4.2. Fourier transform infrared spectrometry (FTIR)

The FTIR spectra for the starch-based bioplastic films were analyzed using a FTIR spectrometer (Bruker Alpha Eco-ATR,
Germany). The transmission of the bioplastic films was obtained between 4000 cm−1 to 600 cm−1 wavenumbers. Each
sample was tested in duplicate for the confirmation of the spectrum (Bhuyar et al., 2019).

2.4.3. Contact angle measurement

The contact angle of each starch-based bioplastic film was analyzed using a drop shape analyzer (KRUSS, DSA25,
Germany). The static contact angle measurement by sessile drop method was done via dispensing distilled water droplet
of 2 µL at room temperature of 30 ◦ C with a relative humidity of 37.5 ± 5%. Six readings were taken for each condition.

2.5. Mechanical characterization

2.5.1. Differential scanning calorimeter (DSC)

Thermal properties of isolated starch were performed by using a differential scanning calorimeter (Netzsche DSC 200F3,
Germany) with a temperature range of 20–100 ◦ C and heating rate of 5 ◦ C per minute under nitrogen (N2 ) atmosphere
at a rate of 20 mL per min. The gelation temperature for the native starch can be predicted from the DSC thermogram.

2.5.2. Tensile test

Starch films’ tensile properties were performed using a universal testing machine (UTM), Instron E1000 (Instron Ltd.
U.K.), following ASTM D3039. Dry samples strips were cut for each starch-based bioplastic film (50 × 12.5 mm) carefully
without rough edges. The thicknesses of samples were measured using a screw gauge before testing. Force (N) and
deformation (mm) were measured using a 250 N load cell at a testing speed of 10 mm/min−1 . The samples were tested
in triplicates.

2.6. In-vitro biological and chemical characterization

2.6.1. Soil biodegradation

Garden soil mixture consisted of soil, cow compost, plant waste, composting seeds, and water. It was dispensed into
plastic dishes (42 × 32 × 12 cm) up to 4 cm in height. The weight of each composition (2 × 2 cm) of films were weighed
and buried at a depth of 2 cm. The plastic dishes were placed under environmental temperature (32 ± 5 ◦ C) at a relative
humidity of 37 ± 5%. The weight of the sample was measured regularly after 24 h of time interval, and its degradation
(weight loss) percentage was calculated using Eq. (1) (Ruggero et al., 2019).
wi − wf
Degradation (%) = × 100 (1)
wi
where wf = final weight after degradation, wi = initial weight before degradation.

2.6.2. Water degradation and swelling test

The swelling capacity of the starch-based bioplastic films was measured in DW. The dry samples were cut from each
film (2 × 2 cm), weighed and immersed in DW at 37 ◦ C for 24 h. The samples were taken out, dried, and weighed. The
swelling capacity was calculated by using the following Eq. (2) (Othman et al., 2017).
wf − wi
Swelling (%) = × 100 (2)
wi
where wf = final weight after swelling, wi = initial weight before swelling.
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

2.6.3. Water vapor transmission rate (WVTR)

The starch films were cut into discs of 1.6 cm diameter to determine the water vapor transmission rate (WVTR). Each
vial contained 10 mL of DW, and a suitable adhesive was used to seal the bioplastic films. The complete setup was weighed
and kept at room temperature for 24 h. The WVTR was calculated using Eq. (3) (Basha et al., 2011).
( g )
WVTR m2
= (wi − wf )/24 ∗ A ∗ 10000 (3)
h
where wi = initial weight before 24 h, wf = final weight after 24 h, and A = area of the films

2.6.4. Solubility test

The dry films were cut (2 × 2 cm), weighed, and immersed into 50 mL of DW. Later, the glass beaker was placed on the
magnetic stirrer for slow agitation at 100 rpm for 24 h. After 24 h, the non-solubilized samples were stripped away, and
the dry mass weight was measured at room temperature. Tests were performed in triplicates to determine the solubility
of the starch films. Solubility was calculated using Eq. (4) (Santana et al., 2018).
mi − mf
Solubility (%) = ∗ 100 (4)
mi
where mi = initial mass of the film, mf = final mass of the non-solubilized film.

2.6.5. Acid, alkali, and salt test

For acid test, bioplastic films were treated with 10% of acetic acid. The dry samples were cut from each composition
film (2 × 2 cm), weighed, and placed into the 20 mL of acid solution. The films were taken out, dried, and its weight was
measured at regular interval. In addition, the alkali behavior of starch-based films was determined by treating them with
10% of sodium hydroxide. The dry samples were cut 2 × 2 cm, weighed and immersed in 20 mL of alkali solution. The
samples were taken out carefully, dried, and its weight was measured. Similarly, the biofilms were treated with 20 mL
of 10% sodium chloride solution for the salt test. The samples were taken out carefully, dried, and weighted until it was
not annihilated (Mostafa et al., 2018).

2.7. Statistical analysis

The obtained results were statistically analyzed using one-way single factor ANOVA. An estimation of *p < 0.05 was
used to validate the significant differences.

3. Results and discussion

3.1. Physicochemical and mechanical characterization

3.1.1. SEM analysis

SEM micrographs depicted the surface morphology of yam starch powder and starch-based bioplastic films in Fig. 2. The
SEM micrograph of yam starch powder (Fig. 2(a)) showed an ellipsoid and oval-shaped structure (Dereje, 2021). According
to the image J calculation, the average particle size of yam starch powder was found to be 32 ± 5.6 µm. The reported value
is in accordance with the previously reported particle size of 8.24 to 48.95 µm (Wu et al., 2021). In non-reinforced (B0)
film, small pore-like structures was observed (as shown in Fig. 2(b)) with the starch granules been thoroughly dissolved
with no residues. There was no significant improvement with the addition of 0.5% bentonite. However, it was observed
that the pores were filled and its surface is smoothened for 1 and 1.5% of bentonite concentration. The existence of pores
and void may affect the mechanical strength such as modulus of elasticity and tensile strength of bioplastic film (Gaaz
et al., 2015; Jayakumar et al., 2021).

3.1.2. DSC analysis

A DSC thermogram investigated the thermal behavior of yam starch powder, as shown in Fig. 3(a). The thermogram
of native yam starch powder indicates its single glass transition temperature (Tg ) at 80 ◦ C, ultimately providing gelation
temperature. Since Tg is above room temperature, it can show glass-like behavior in strength, brittleness, stiffness, etc.
(Smith et al., 2009). A small peak observed around 22 ◦ C. It may be due to the presence of two different morphologies in
yam starch powder, as shown in SEM micrographs Fig. 2(a) (Wang et al., 2008). The thermal behavior plays a vital role in
determining the physicomechanical behavior of polysaccharides films as it persuades the amorphous/ crystalline phase.
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

Fig. 2. SEM micrographs of (a) yam starch powder, (b) B0, (c), B1, (d) B2, and (d) B3 bioplastic films.

3.1.3. Tensile testing

The effect of bentonite reinforced filler concentration on the tensile strength (Fig. 3(b)) of the starch-based bioplastic
films was analyzed using a UTM. Santana et al. (2018) had reported that the increase in tensile strength is only up to 5%
of starch after that it starts decreasing. Also, 1.5% glycerol was found to be optimum after that its plasticizing property
decreases. Earlier studies reported, 1.5% concentration of bentonite in composite material was found to have optimum
mechanical strength. Beyond 1.5% concentration of bentonite, the mechanical strength was found to be decrease (Luckham
and Rossi, 1999; Kar et al., 2016). Shanmathy et al. (2021) also reported that the bentonite reinforced bioplastic film
produced from 3% starch has good tensile strength. Accordingly, it was observed from Table 2 that with the increase in
the concentration of bentonite, the tensile strength and Young’s modulus increased. Additionally, it was observed that
energy at break was higher for B2 film. Bentonite concentration improves the mechanical properties of bioplastic films. It
might be due to the homogeneous dispersion of bentonite, in the bioplastic composite film. The concentration of bentonite
cannot be increased beyond a particular quantity due to the agglomeration of clay particles and subsequent reduction in
the mechanical strength of the bioplastic film (Luckham and Rossi, 1999). As shown in SEM micrographs, the pores are
almost filled with an increase in bentonite concentration which shows that bentonite filler improves the film’s strength. It
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

Fig. 3. Graphical representation of (a) DSC of yam starch powder, (b) Tensile strength, (c) FTIR spectra, and (d) Contact angle of B0, B1, B2, and B3
bioplastic films. Note: *p < 0.05 and **p < 0.005 comparative to control.

Table 2
Calculated mechanical parameters of yam starch-based bioplastic films.
Sample Tensile Strength (MPa) Young’s Modulus (MPa) Energy at break (J)
B0 2.56 ± 0.13 40.9 ± 6.04 0.03 ± 0.005
B1 2.66 ± 0.72 45.35 ± 3.03 0.03 ± 0.013
B2 3.053 ± 0.361 54.38 ± 7.89 0.063 ± 0.030
B3 4.063 ± 0.12 72.65 ± 11.42 0.06 ± 0.009

might be due to the formation of an H2 bond between the yam starch and intercalated layered configuration of bentonite
(Rhim et al., 2006; Natália and Cristina, 2010). The consistency of matrix in films is a good indicator of their structural
reliability, and consequently, good mechanical properties would be expected (Zamrud et al., 2021).

3.1.4. FTIR analysis

The FTIR spectra of B0, B1, B2, and B3 bioplastic films are represented in Fig. 3(c). It was observed that yam starch
powder does not form a peak between 860 to 900 cm−1 , but with the addition of glycerol in B0, B1, B2, and B3 bioplastic
film, carbohydrate vibration peak was observed in this region. On the other hand, the siloxane (Si–O–Si) functional group
of bentonite gets more prominent with the increase in its concentration at 1000 cm−1 for B1, B2, and B3 bioplastic films
(Foletto et al., 2003). The band at 1200 cm−1 of a starch group (C=O) was significantly observed for all bioplastic films.
The prominent peak of the C–H stretch was observed for B0, B1, B2, and B3 (Coates, 2000). The bands between 3000–
3500 cm−1 relate with the –OH group due to hydroxyl bunches of water molecules present between the layer space of
bentonite powder on B1, B2, and B3 (Zhang et al., 2011; Bhuyar et al., 2019). The broadening of the peak intensity was
observed at 1640 cm−1 is due to the addition of bentonite in the taro starch matrix, which indicates the interaction of
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

the aluminum (Al) group from bentonite with the OH group (Abdullah and Dong, 2019). Thus, these following functional
groups, such as OH and CO ester groups, suggest that the taro starch-based films have excellent biodegradability properties
(Rusianto et al., 2019).

3.1.5. Contact angle measurement

Hydrophilicity plays an essential role in the interaction between the particles. The measured contact angles for B0,
B1, B2, and B3 films were 80.55 ± 7.35, 56.2 ± 4.4, 48.55 ± 4.15, and 48.9 ± 1.2 respectively. A decrease in the contact
angle was observed with the bentonite concentration due to the presence of more negative groups on the surface (O–H
group), as shown in FTIR spectra (Fig. 3(d)). Even though there are no significant differences between the contact angle
values, there is a trend towards hydrophilicity in the film containing bentonite compared to the B0 bioplastic film. The
hydrophilicity of starch-based bioplastic film containing bentonite (B1, B2, and B3) enhances, which tends to increase their
water absorption capacity. It is in accordance with the Van Oss and Giese (1995) a surface thermodynamic hypothesis,
that the Na+ -montmorillonite surface is hydrophilic, which clarifies the massive take-up of water between the platelets
bentonite clay mineral.

3.2. In-vitro biological testing

3.2.1. Soil degradation test

Soil degradation analysis was done by utilizing garden soil, and the weight of the bioplastic films was estimated each
day. From Fig. 4(a), it is seen that the bioplastic film without bentonite (B0) had a slower degradation initially; however,
it debased quickly after 50 days. It was observed that the biodegradable nature of bentonite enhances the degradation
rate of starch-based bioplastic films. Sample B3 (1.5% bentonite) was found with maximum degradation among all starch
composite films. Due to the intercalated-layered silicates of bentonite, which were dispersed in the yam starch-glycerol
grid, it develops a good contact between the network and bentonite surface (Guo et al., 2018). This outcome remains as
proof to demonstrate that the bioplastic films were biodegradable and ecological amicable.

3.2.2. Water degradation and swelling test

Water absorption capacity is the critical characteristic of biodegradable bioplastic. The B0 bioplastic film took nearly
about 15 days to degrade in water. Whereas B1, B2, and B3 starch-based bentonite contained bioplastic film took 20–25
days for complete degradation in water (Fig. 4(b)). The bentonite films absorb moisture gradually, which makes them
degrade slowly in water. Therefore, yam starch composites are water tainted without harming environmental conditions.
It was observed that B1, B2, and B3 films swelling pace was slow due to bentonite’s presence; its potential for absorbing
moisture enhances and limits their swelling rate compared to B0 film, (Fig. 4(c)). It was accounted for that in DW, zeta
capability of bentonite is negative over whole pH 4–10 because of transport of H+ particle from clay to water. Thus, as
a grouping of bentonite builds, the swelling behavior of bioplastic film is brutal to notice (Niriella and Carnahan, 2006).
This swelling detaches the bentonite particle, particularly those arranged in parallel direction until they attain equilibrium
(Chen et al., 1987).

3.2.3. Water vapor permeability test

Water vapor permeability (WVP) is a way to measure water vapor channels across a film membrane. It plays a
vital role in food packaging to avert moisture transmission from the neighboring environment to the food. Also, water
vapor permeability is directly proportional to the water vapor transmission rate. From Fig. 4(d), it was observed that B0
without bentonite has a higher water vapor transmission rate than B1, B2, and B3. The measured WVTR values of B0,
B1, B2, and B3 are 36.79 ± 0.11 × 10−6 kg/m2 day (0.036 g/m2 day), 35.52 ± 0.11 × 10−6 kg/m2 day (0.035 g/m2 day),
32.55 ± 0.11 × 10−6 kg/m2 day (0.032 g/m2 day), and 31.07 ± 0.12 × 10−6 kg/m2 day (0.031 g/m2 day), respectively.
It might be due to the starch arrangement formed by the plasticizer (glycerol), and its hydrophilic property supports the
inclusion and desorption of water molecules. The increase in bentonite concentration leads to a decrease in the water
vapor transmission rate. An increase in the film thickness leads to an increase in water resistance. Thus, it creates a
fractional equilibrium pressure of water on the interior surface of the film. Some of the previously described WVTR values
for food packaging materials such as illite-PE composite films, thin-film coated PMP, and PLA films are 13.27 g/m2 day,
0.57 g/m2 day, 21 g/m2 day, respectively (Wu et al., 2020; Seong et al., 2020; Shogren, 1997). Recommended values
of WVTR for wrapping materials are between 0.01 to 1 g/m2 day (Spee et al., 2015). Therefore, the developed yam
starch-based bioplastic films satiate this requirement with the increasing concentration of bentonite.

3.2.4. Solubility test

Solubility percentage increased with increasing bentonite concentration, as shown in Fig. 5(a). Intermolecular attraction
forces between bentonite molecules are meagre, enabling them to absorb more water and make film more solubilization
(Naidu and John, 2020). Higher solubility of the film affects the commercial purpose of the film because aqueous or liquid
food ingests the film. The solubility of B3 film in water was high due to the weak intermolecular force.
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

Fig. 4. Graphical representation of (a) Soil biodegradation, (b) Water degradation, (c) Swelling ratio, and (d) Water vapor transmission rate of B0,
B1, B2, and B3 bioplastic films. Note: *p < 0.05 comparative to control.

3.3. Chemical testing

3.3.1. Acid, alkali, and salt test

The acid, alkali, and salt test measures the time duration of starch-based bioplastic film to withstand the chemical
medium, which is beneficial for food packaging application. It was observed from Fig. 5(b) that B0 bioplastic film resists in
a 10% acidic medium (pH 4) up to 36 h, whereas B1, B2, and B3 bioplastic film took almost 40 h to disintegrate in the acidic
medium. It may be due to the reduction of cohesiveness in starch-based films. In B1, B2, and B3 bioplastic films, bentonite
(montmorillonite) dissolution occurs in acetic acid medium, leading to the removal of interlayer cations. Therefore, it
increases the pore volume and ultimately increases bentonite adsorption properties. The starch-based bioplastic films
swell rapidly within 20 min in 10% NaOH solution at pH 9, as demonstrated in Fig. 5(c) (Shanmathy et al., 2021; Mohanta
and Thirugnanam, 2021). It was reported that interlayer magnesium (Mg+) ions in the bentonite structure are prominently
affected by the high sodium (Na) content solution (Rosário et al., 2010). Similarly, starch-based bioplastic films were
immersed in 10% NaCl solution (shown in Fig. 5(d)). It was observed that films remained integrated and steady in the
salt solution. The increase in bentonite has a significant impact on the swelling behavior of the film. With the rise in the
bentonite concentration, swelling properties increases, but its weight remains almost stable up to 28 h. This is due to the
salt solution’s low charge impact from the edge site (Erdemoğlu, 2007). Consequently, bentonite particle does not exhibit
any variation in the surface charge.

4. Conclusion

Yam starch-based bioplastic film with and without bentonite has been successfully developed by using the solvent
casting method. The bentonite concentration significantly affected the properties of the bioplastic film. It was observed
from SEM micrographs that bentonite was homogeneously distributed in the starch matrix, which eliminates all voids,
and the surface got smoothened completely. This consistency of matrix leads to an increase in the tensile strength of
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

Fig. 5. Graphical representation of (a) Solubility, (b) Acid test, (c) Alkali test, and (d) Salt test of B0, B1, B2, and B3 bioplastic films. Note: *p < 0.05
comparative to control.

starch-based bioplastic film. Further, it was observed that B3 bioplastic exhibited highest soil degradation due to bentonite
biodegradable nature, which makes it degrade entirely within 55 days. For degradation in water, starch-based bentonite
(B1, B2, and B3) bioplastic films took only 20–25 days to degrade. All the bioplastic films were ineffective against NaOH,
while they stayed stable in salt and acidic medium. Similarly, the decrease in WVTR encounters the prime requirement
of food packaging. Therefore, it is well inferred that yam starch with bentonite reinforced filler can create bioplastic
films for food packaging applications. Further studies can be done with different filler additives to improve mechanical
properties as well as to increase overall shelf life of the bioplastic films. Moreover, different types of plasticizers can be
utilized to evaluate the physical, mechanical and environmental properties of the produced yam starch-based bioplastic
for packaging application.

CRediT authorship contribution statement

Lopamudra Behera: Conceptualization, Methodology. Monalisha Mohanta: Data curation, Writing – original draft.
Arunachalam Thirugnanam: Supervision, Visualization, Investigation, Validation.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have
appeared to influence the work reported in this paper.

Acknowledgment

The authors thank the Department of Biotechnology and Medical Engineering, National Institute of Technology
Rourkela, India, for providing research facilities.
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L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

References

Abdullah, Z.W., Dong, Y., 2019. Biodegradable and water resistant poly(vinyl) alcohol (PVA)/starch (ST)/glycerol (GL)/halloysite nanotube (HNT)
nanocomposite films for sustainable food packaging. Front. Mater. 6 (58), http://dx.doi.org/10.3389/fmats.2019.00058.
Acquavia, M.A., Pascale, R., Martelli, G., Bondoni, M., Bianco, G., 2021. Natural polymeric materials: A solution to plastic pollution from the agro-food
sector. Polymers 13 (158), http://dx.doi.org/10.3390/polym13010158.
Agarwal, S., 2021. Major factors affecting the characteristics of starch-based biopolymer films. Eur. Polym. J. 160, 110788. http://dx.doi.org/10.1016/
j.eurpolymj.2021.110788.
Alekseeva, O.V., Rodionova, A.N., Bagrovskaya, N.A., Agafonov, A.V., Noskov, A.V., 2017. Effect of the organobentonite filler on structure and properties
of composites based on hydroxyethyl cellulose. J. Chem. 2017 (11), http://dx.doi.org/10.1155/2017/1603937.
Basha, R.K., Konno, K., Kani, H., Kimura, T., 2011. Water vapor transmission rate of biomass-based film materials. EAEF 4, 37–42. http://dx.doi.org/
10.1016/S1881-8366(11)80018-2.
Bhuyar, P., Muniyasamy, S., Govindan, N., 2018. Green revolution to protect environment – an identification of potential micro algae for the
biodegradation of plastic waste in Malaysia. Expert Opin. Environ. Biol. 7, 87. http://dx.doi.org/10.4172/2325-9655-C5-033.
Bhuyar, P., Tamizi, N.A.B.M., Rahim, Maniam, G.P., Govindan, N., 2019. Effect of ultraviolet light on the degradation of low-density polyethylene
characterized by the weight loss and FTIR. MIJEEC 1, 26–31. http://dx.doi.org/10.54279/mijeec.v1i2.244915.
Bhuyar, P., Trejo, M., Dussadee, N., Unpaprom, Y., Ramaraj, R., Whangchai, K., 2021. Microalgae cultivation in wastewater effluent from tilapia culture
pond for enhanced bioethanol production. Water Sci. Technol. 2–8. http://dx.doi.org/10.2166/wst.2021.194.
Bodiralu, R., Teaca, C.A., Spiridon, L., 2013. Influence of natural fillers on the properties of starch-based biocomposite films. Compos. B. Eng 44,
575–583. http://dx.doi.org/10.1016/j.compositesb.2012.02.039.
Chen, S., Low, P.F., Cushman, J.H., Roth, C.B., 1987. Organic compound effects on swelling and flocculation of upton montmorillonite. Soil Sci. Soc.
Am. J 51, 1444–1450, http://docs.lib.purdue.edu/watertech/178.
Coates, J., 2000. Interpretation of Infrared Spectra, a Practical Approach. Encyclopedia of Analytical Chemistry. John Wiley & Sons Ltd, Chichester, pp.
10815–10837. http://dx.doi.org/10.1002/9780470027318.a5606.
Dereje, B., 2021. Composition, morphology and physicochemical properties of starches derived from indigenous Ethiopian tuber crops: A review. Int.
J. Biol. Macromol. 187, 911–921. http://dx.doi.org/10.1016/j.ijbiomac.2021.07.188.
Dos Santos, R.G., dos Santos, R.A., Teles, D., Andrades, R., Martins, A.S., 2019. Scavenging as a pathway for plastic ingestion by marine animals.
Environ. Pollut. 248, 159–165. http://dx.doi.org/10.1016/j.envpol.2019.02.010.
Dos Santos Caetano, K., Almeida Lopes, N., Haas Costa, T.M., Brandelli, A., Rodrigues, E., Hickmann Flôres, S., Cladera-Olivera, F., 2018. Characterization
of active biodegradable films based on cassava starch and natural compounds. Food Packag. Shelf Life. 16, 138–147. http://dx.doi.org/10.1016/j.
fpsl.2018.03.006.
Erdemoğlu, M., 2007. Zeta potential of pyrophyllite in aqueous solutions of alkaline and alkaline earth metal cations and low-molecular-weight
organic anions. J. Dispers. Sci. Technol. 28, 689–695. http://dx.doi.org/10.1080/01932690701341793.
Eriksson, O., Finnveden, G., 2009. Plastic waste as a fuel – co2 -neutral or not? Energy Environ. Sci. 2, 907–914. http://dx.doi.org/10.1039/b908135f.
Foletto, E.L., Volzone, C., Porto, L.M., 2003. Performance of an argentinian acid-activated bentonite in the bleaching of soybean oil. Food science
engineering. Braz. J. Chem. Eng. 20, http://dx.doi.org/10.1590/S010466322003000200007.
Gaaz, T.S., Sulong, A.B., Akhtar, M.N., Kadhum, A.A., Mohamad, A.B., Al-Amiery, A.A., 2015. Properties and applications of polyvinyl alcohol. Halloysite
Nanotub. Nanocomposit Mol. 20, 22833–22847. http://dx.doi.org/10.3390/molecules201219884.
Grigore, M.E., 2017. Methods of recycling, properties, and applications of recycled thermoplastic polymers. Recycling 2410, 2–11. http://dx.doi.org/
10.3390/recycling2040024.
Guo, F., Aryana, S., Han, Y., Jiao, Y., 2018. A review of the synthesis and applications of polymer–nanoclay composites. Appl. Sci 8 (1696),
http://dx.doi.org/10.3390/app8091696.
Hofmann, U., Endell, D., 1933. Kristallstruktur und Quellung von Montmorillonit. (Das Tonmineral der Bentonittone). Z. Krist. 86, 340–348.
http://dx.doi.org/10.1524/zkri.1933.86.1.340.
Jadhav, P., Muhammad, N., Bhuyar, P., Krishnan, S., Razak, S., Abd, A.W., Zularisam, A.W., Mohd. Nasrullah, M., 2021. A review on the impact of
conductive nanoparticles (CNPs) in anaerobic digestion: Applications and limitations. Environ. Technol. Innov. 23, 101526. http://dx.doi.org/10.
1016/j.eti.2021.101526.
Jayakumar, S., Bhuyar, P., Pugazhendhi, A., Rahim, Maniam, G.P., Govindan, N., 2021. Effects of light intensity and nutrients on the lipid content
of marine microalga (diatom) Amphiprora sp. for promising biodiesel production. Sci. Total Environ. 768, 145471. http://dx.doi.org/10.1016/j.
scitotenv.2021.145471.
Kar, S., Kaur, T., Thirugnanam, A., 2016. Microwave-assisted synthesis of porous chitosan-modified montmorillonite-hydroxyapatite composite
scaffolds. Int. J. Biol. Macromol 82, 628–636. http://dx.doi.org/10.1016/j.ijbiomac.2015.10.060.
Khazaai, S.N.M., Bhuyar, P., Rahim, Alwi, M.H.F.M., Yiting, S., Maniam, G.P., 2021. Rapid determination of diesel/biodiesel blend ratio using refractive
index, density, and kinematic viscosity measurements. Biomass Conv. Bioref. http://dx.doi.org/10.1007/s13399-021-01921-z.
King, A., Singh, R., Nayak, B.B., 2021. Phase and photoluminescence analysis of dual-color emissive Eu3 + doped ZrO2 nanoparticles for advanced
security features in anti-counterfeiting. Colloids Surf. A Physicochem. Eng. Asp 631, 127715. http://dx.doi.org/10.1016/j.colsurfa.2021.127715.
Luckham, P.F., Rossi, S., 1999. The colloidal and rheological properties of bentonite suspensions. Adv. Colloid Interface Sci. 82, 43–92. http:
//dx.doi.org/10.1016/S0001-8686(99)00005-6.
Mali, S., Grossmann, M.V.E., Garcia, M.A., Martino, M.N., Zaritzky, N.E., 2005. Mechanical and thermal properties of yam starch films. Food Hydrocoll.
19, 157–164. http://dx.doi.org/10.1016/10.1016/j.foodhyd.2004.05.002.
Marsh, K., Bugusu, B., 2007. Food packaging—Roles, materials, and environmental issues. J. Food Sci. 72, R39–R55. http://dx.doi.org/10.1111/j.1750-
3841.2007.00301.x.
Mohanta, M., Thirugnanam, A., 2021. Drug release studies of titanium-based polyethylene glycol coating as a multifunctional substrate. Mater. Today
Proc. 47, 257–260. http://dx.doi.org/10.1016/j.matpr.2021.04.242.
Mostafa, N.A., Farag, A.A., Abo-dief, H.M., Tayeb, A.M., 2018. Production of biodegradable plastic from agricultural wastes. Arabian J. Chem. 11,
546–553. http://dx.doi.org/10.1016/j.arabjc.2015.04.008.
Naidu, D.S., John, M.J., 2020. Effect of clay nanofillers on the mechanical and water vapor permeability properties of Xylan–Alginate films. Polymers
12 (2279), http://dx.doi.org/10.3390/polym12102279.
Natália, F.M., Cristina, T.A., 2010. Calcium bentonite as reinforcing nanofiller for thermoplastic starch. J. Brazil Chem. Soc. 21, 202–208. http:
//dx.doi.org/10.1590/S0103-50532010000200003.
Navarro, D., Abelilla, J.J., Stein, H.H., 2019. Structures and characteristics of carbohydrates in diets fed to pigs: a review. J. Anim. Sci. Biotechnol. 10,
39. http://dx.doi.org/10.1186/s40104-019-0345-6.
Niriella, D., Carnahan, R.P., 2006. Comparison study of zeta potential values of bentonite in salt solutions. J. Dispers. Sci. Technol 27, 123–131.
http://dx.doi.org/10.1081/DIS-20006686010.1081/DIS-200066860.

11
L. Behera, M. Mohanta and A. Thirugnanam Environmental Technology & Innovation 25 (2022) 102180

Omotoso, M.A., Ayorinde, G.O., Akinsanoye, O.A., 2015. Preparation of biodegradable plastic filmsfrom tuber and root starches. IOSR-JAC 8 (4), 10–20.
http://dx.doi.org/10.9790/5736-08411020.
Othman, S.H., Edwal, S.A.M., Risyon, N.P., Basha, R.K., Talib, R.A., 2017. Water sorption and water permeability properties of edible film made from
potato peel waste. Food Sci. Technol. 37, 63–70. http://dx.doi.org/10.1590/1678-457X.30216.
Peixoto, D., Pinheiro, C., Amorim, J., Oliva-Teles, L., Guilhermino, L., Vieira, M.N., 2019. Microplastic pollution in commercial salt for human
consumption: A review. Estuar. Coast. Shelf Sci. 219, 161–168. http://dx.doi.org/10.1016/j.ecss.2019.02.018.
Rhim, J.W., Hong, S.I., Park, H.M., Ng, P.K.W., 2006. Preparation and characterization of chitosan-based nanocomposite films with antimicrobial activity.
J. Agric. Food Chem. 54, 5814–5822. http://dx.doi.org/10.1021/jf060658h.
Rosário, J.A., Silva, L.A., Moura, G.B.G., Lima, R.B., Brys, M.E.P., Kuhnen, N.C., 2010. Influence of alkaline activation over swelling and cation exchange
capacity on bentonites. Mater. Sci. Forum. 660–661, 1064–1069. http://dx.doi.org/10.4028/www.scientific.net/MSF.660-661.1064.
Rugchati, O., Thanacharoenchanaphas, K., 2015. Application of biodegradable film from Yam (Dioscorea alata) Starch in Thailand for agriculture
activity. Int. J. Environ. Rural Dev. 6, 28–33, http://iserd.net/./IJERD%2016-1-05.pdf.
Ruggero, F., Gori, R., Lubello, C., 2019. Methodologies to assess biodegradation of bioplastics during aerobic composting and anaerobic digestion: A
review. WM & R 37, 959–975. http://dx.doi.org/10.1177/0734242X19854127.
Rusianto, T., Yuniwati, M., Wibowo, H., Akrowiah, L., 2019. Biodegradable Plastic from Taro Tuber (Xanthosoma sagittifolium) and Chitosan. Saudi J.
Eng. Technol. 4 (1), 16–22. http://dx.doi.org/10.21276/sjeat.2019.4.1.3.
Santana, R.F., Bonomo, R., Gandolfi, O., Rodrigues, L.B., Santos, L.S., Dos Santos Pires, A.C., de Oliveira, C.P., da Costa Ilhéu Fontan, R., Veloso, C.M.,
2018. Characterization of starch-based bioplastics from jackfruit seed plasticized with glycerol. J. Food Sci. Technol. 55, 278–286. http://dx.doi.
org/10.1007/s13197-017-2936-6.
Seong, D.M., Lee, H., Kim, J., Chang, J.H., 2020. High oxygen and water-vapor transmission rate and in vitro cytotoxicity assessment with
illite-polyethylene packaging films. Materials 13 (2382), http://dx.doi.org/10.3390/ma13102382.
Shanmathy, M., Mohanta, M., Thirugnanam, A., 2021. Development of biodegradable bioplastic films from taro starch reinforced with bentonite.
Carbohydr. Polym. Technol. Appl. 2, 100173. http://dx.doi.org/10.1016/j.carpta.2021.100173.
Shogren, R., 1997. Water vapor permeability of biodegradable polymers. J. Polym. Environ. 5, 91–95. http://dx.doi.org/10.1007/BF02763592.
Smith, K.E., Sawicki, S., Hyjek, M.A., Downey, S., Gall, K., 2009. The effect of the glass transition temperature on the toughness of photopolymerizable
(meth)acrylate networks under physiological conditions. Polymer 50, 5112–5123. http://dx.doi.org/10.1016/j.polymer.2009.08.040.
Souza, A.C., Benze, R., Ferrão, E.S., Ditchfield, C., Coelho, A.C.V., Tadini, C.C., 2012. Cassava starch biodegradable films: Influence of glycerol
and clay nanoparticles content on tensile and barrier properties and glass transition temperature. LWT - Food Sci. Technol. 46, 110–117.
http://dx.doi.org/10.1016/j.lwt.2011.10.018.
Spee, D.A., Rath, J.K., Schropp, R.E.I., 2015. Using hot wire and initiated chemical vapor deposition for gas barrier thin-film encapsulation. Thin Solid
Films 575, 67–71. http://dx.doi.org/10.1016/j.tsf.2014.10.029.
Sukhija, S., Singh, S., Riar, C.S., 2015. Isolation of starches from different tubers and study of their physicochemical, thermal, rheological and
morphological characteristics. Starke 67, 1–9. http://dx.doi.org/10.1002/star.201500186.
Sundararaju, S., Arumugam, M., Bhuyar, P., 2020. Microbacterium sp. MRS-1, a potential bacterium for cobalt reduction and synthesis of less/non-toxic
cobalt oxide nanoparticles (Co3 O4 ). Beni-suef univ. J. Basic Appl. Sci. 9, 44. http://dx.doi.org/10.1186/s43088-020-00070-y.
Talsness, C.E., Andrade, A.J., Kuriyama, S.N., Taylor, J.A., Vom Saal, F.S., 2009. Components of plastic: experimental studies in animals and relevance
for human health. Philos. Trans. R. Soc. Lond. B. Biol. Sc 364, 2079–2096. http://dx.doi.org/10.1098/rstb.2008.0281.
Ulyarti, U., Lavilinesia, L., Surhaini, S., Siregar, N., Tomara, A., Lisani, L., Nazarudin, N., 2021. Development of yam starch-based bioplastics with the
addition of chitosan and clove oil. Makara J. Sci. 25, 91–97. http://dx.doi.org/10.7454/mss.v25i2.1155.
Van Oss, C.J., Giese, R.F., 1995. The hydrophilicity and hydrophobicity of clay minerals. Clays. Clay Miner. 43, 474–477. http://dx.doi.org/10.1346/
CCMN.1995.0430411.
Vu, Nguyen., H.P, Lumdubwong, N., 2016. Starch behaviors and mechanical properties of starch blend films with different plasticizers. Carbohydr.
Polym. 154, 1–36. http://dx.doi.org/10.1016/j.carbpol.2016.08.034.
Wang, X., Gao, W.Y., Zhang, L., Xie, W., 2008. Study on the morphology, crystalline structure, and thermal properties of yam starch acetates with
different degrees of substitution. Sc. China Ser B Chem. 51, 859–865. http://dx.doi.org/10.1007/s11426-008-0089-1.
Wu, Z., Ameer, K., Hu, C., Bao, A., Wang, R., Tang, W., Chaudhary, N., Jiang, G., 2021. Particle size of yam flour and its effects on physicochemical
properties and bioactive compounds. Food Sci. Technol. 1–9. http://dx.doi.org/10.1590/fst.43921.
Wu, H.Y., Liu, T.X., Hsu, C.H., Cho, Y.S., Xu, Z.J., Lia, S.C., Zeng, B.H., Jiang, Y.L., Lien, S.Y., 2020. High oxygen and water-vapor transmission rate and
in-vitro cytotoxicity assessment with illite-polyethylene packaging films. Materials 13 (2382), http://dx.doi.org/10.3390/ma13102382.
Zamrud, Z., Ng, W.M., Salleh, H.M., 2021. Effect of bentonite nanoclay filler on the properties of bioplastic based on sago starch. IOP Conf. Ser. Earth
Environ. Sci. 765, 012009. http://dx.doi.org/10.1088/1755-1315/765/1/012009.
Zhang, L., Yang, X., Li, S., Gao, W., 2011. Preparation, physicochemical characterization and in vitro digestibility on solid complex of maize starches
with quercetin. J. Food Sci. Technol. 44, 787–792. http://dx.doi.org/10.1016/j.lwt.2010.09.001.

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