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OPEN New polyvinyl alcohol/gellan

gum‑based bioplastics with guava
and chickpea extracts for food
packaging
Shaimaa Elsaeed 1*, Elsayed Zaki 1, Ayman Diab 2, Menna‑Alla Tarek 2 & Walaa A. E. Omar 3*
Plastic is a fossil-based synthetic polymer that has become an essential material in our daily life.
Plastic pollution resulting from the accumulation of plastic objects has become problematic for our
environment. Bioplastic can be a biodegradable environmentally friendly alternative for the synthetic
plastic. In this paper, bioplastics based on polyvinyl alcohol (PVA)/gellan gum (GG) blend have been
produced in three different compositions and their chemical structure, mechanical, morphological and
thermal properties have been studied. Glycerol has been used as a plasticizer. To add extra features
to the PVA/GG bioplastic, Psidium guajava (guava) leaves, GL, and chickpea, CP, extracts have been
added to the PVA/GG (30/70) blend. Water and aqueous ethanol have been used in the extraction of
GL and CP, respectively. The addition of the plant’s extracts enhanced the tensile properties of the
PVA/GG bioplastic. Weathering acceleration tests have been carried out to examine the degradation
of the prepared bioplastics. Cytotoxicity studies revealed that the prepared bioplastic is safe to be
used in food packaging applications. Water and oxygen permeability for the new PVA/GG bioplastic
have also been studied. The addition of the plant extracts (GL and CP extracts) increased the oxygen
and water permeability to different extents. Bioplastic life cycle assessment (LCA) and ­CO2 emissions
in comparison to fossil-based plastic have been investigated. From all the results, PVA/GG based
bioplastic proved to be a degradable, safe and effective alternative for fossil-based plastics in food
packaging applications.

Plastics are petroleum based organic polymers that possess extraordinarily rapid growth and large-scale produc-
tion due to their lightweight, chemical stability and low processing c­ ost1–3. This massive production is derived
by the global shift from reusable to single use containers. On the other hand, the huge plastic production gen-
erated a global solid waste accumulation that is growing steadily over the ­years4. There are two basic classes of
plastics, thermoplastics and thermosets. For both classes of plastics, most of the monomers used to produce
them are derived from fossil hydrocarbons such as ethylene, propylene, and styrene. Therefore, plastics are non-
biodegradable i.e., they don’t decompose naturally. Consequently, plastic accumulates in the earth and oceans
causing serious environmental and health p ­ roblems1–5 .
In contrast to plastics, bioplastics includes polymers that are biodegradable and/or originate from renew-
able ­resources2,6. They can be classified into four classes, Class (I) which includes the polysaccharides that are
directly extracted from biomass such as ­starch7,8, xanthan ­gum9, guar ­gum10, gellan gum and proteins. Class (II)
of bioplastics includes materials that are bio-sourced but not biodegradable such as polyethylene produced from
ethanol by ­fermentation2. Class (III) of bioplastics includes materials that are produced from petrochemicals but
are degradable. An example of such materials is polyvinyl alcohol (PVA, the structure is illustrated in Fig. 1a).
Class (IV) are polymers that are produced by the action of bacteria such as ­polyhydroxyalkanoate2.
Bioplastics made from blends of two or more types of the above-mentioned polymers have been a target of
many research groups aiming to have enhanced/desired mechanical properties and/or lower c­ ost2,11,12.
Polyvinyl alcohol (PVA, Fig. 1a)1, which belongs to class (III) in bioplastics ­classification2, is a water-soluble
synthetic biodegradable polymer. It is widely used as thermoplastic polymer and is usually prepared by hydroly-
sis of polyvinyl acetate. PVA is nontoxic and has many applications in industry, medicine, and food sector,

1
Egyptian Petroleum Research Institute, Naser City, Cairo 11727, Egypt. 2Faculty of Biotechnology, October
University for Modern Sciences and Arts, 6th of October City, Egypt. 3Faculty of Petroleum and Mining
Engineering, Suez University, P.O.Box: 43221, Suez, Egypt. *email: [email protected]; walaa.omar@
pme.suezuni.edu.eg

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Figure 1.  (a) The chemical structure of ­PVA1, (b) ­GG20,21, and (c) Glycerol.

specifically food p ­ ackaging13. Physical mixing of synthetic polymers such as PVA with natural polymers repre-
sents an effective way to obtain polymeric materials with desired features, improved mechanical characteristics
and biological ­performances14. For that purpose, PVA was blended successfully with many natural polymers and
the produced copolymers are successfully employed in many fields. In medicine, PVA blends were promising
candidates in tissue engineering. PVA/chitosan blend has been employed in neural tissues ­therapies15. While
PVA/PET (polyethylene terephthalate) blend has been applied in vascular grafts, PVA/collagen has been used
in corneal applications. PVA/Gellan gum (GG) biodegradable nanofibers have been prepared for skin tissue
­regeneration11,12.
In the same context, films formed from such PVA blends showed enhanced mechanical properties for indus-
trial ­applications16. The films mechanical properties could be tuned by the PVA percentage in the blend. PVA
blends with natural polymers such as guar g­ um17, ­starch1,18, ­cellulose19 and ­chitosan20 have been reported in
food packaging applications. PVA is soluble in water due to the presence of the hydroxyl groups in its backbone
(Fig. 1a)1. The PVA hydrophilicity enables the use of water as a solvent during blending PVA with other polymers.
The water temperature should be around 100 °C to have the PVA completely soluble and ready to be mixed with
other dissolved polymers forming a homogenous ­solution16.
Brunchi et al.14 studied the interaction in the blend PVA /xanthan gum. FTIR analysis proved the formation
of a network of hydrogen bonds between the carboxylate and hydroxyl groups in xanthan gum and the hydroxyl
groups of PVA.
Gellan gum (GG) is a biocompatible thermally stable polysaccharide (Fig. 1b). It is a hydrophilic biopolymer
that is widely recognized as a food a­ dditive21,22. However, it has poor mechanical strength, high gelling tempera-
ture and small temperature window. Therefore, GG was blended with different natural and synthetic polymers
to have a blend with tunable mechanical and physical p ­ roperties11,12.
23
Paolicelli et al. have prepared thin films of plasticized GG by solvent casting technique for oral delivery appli-
cations. They reported that the use of glycerol (Fig. 1c) as a plasticizer by an amount of 20–75% w/w enhanced
the mechanical characteristics of GG based films as it reduced the interconnection among the polymeric chains.
In general, glycerol is a biodegradable, non-toxic trihydroxy compound that is widely used as a p ­ lasticizer24,25.
12
PVA\GG composites have been the target of a few studies. Vashisth et al. prepared electro-spun PVA\Gel-
lan nanofibers. The composite has been employed in biomedical engineering after stabilization by cross-linking
under different conditions.
Sudhamani et al.26 reported PVA/GG blended films by solution casting method. They used water as solvent
and glycerol as a plasticizer. The thermal characteristics, intermolecular interactions and mechanical properties
have been studied by differential scanning calorimetry (DSC), Fourier transfer spectroscopy (FTIR) and tensile
tests, respectively. Although the mechanical properties suggested that films from that blend are potential candi-
dates for food packaging, no extended real performance or hydro-stability tests were provided.
Bioplastic with extra features and properties that suit the packaging purpose would be highly desirable.
Research groups have incorporated plants extract to add value to bioplastics. Tran et al.27 added oregano waste
extract to propylene carbonate/cellulose acetate bioplastic to obtain high UV protection and antioxidant food
packaging. Jung et al.28 mixed a naturally produced indigo pigment with polyhydroxyalkanoate to produce blue
colored bioplastic. Pea protein-based bioplastics have also been produced. This type of bioplastics possessed
enhanced mechanical and antimicrobial p ­ roperties29.

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Among the plants, Psidium guajava (Guava) leaves extract, which is denoted as GL in this paper, has several
biological activities. It has antifungal, antimicrobial, anti-cancer, antioxidant, and antitumor ­properties30–32.
Moreover, the bioactive compounds extracted from guava leaf have been used in the preparation of jelly referred
as guava leaf extract jelly. Adequate intake of these jellies helped the consumer lower high cholesterol levels in
the body and boost up i­ mmunity33. Guava leaves extract have been recently added to sodium alginate to develop
antimicrobial food p ­ ackaging34.
Chickpeas (Cicer arietinum), CP, have got several applications in food, cosmetics, and ­packaging35, in addition
to their applications in medicine such as the treatment of ulcer, bronchitis, cholera, constipation, and ­cancer36,37.
Chickpea aquafaba has also been recently employed in bioplastic p ­ ackaging35. Additionally, the Proteins isolated
from the Chickpea seeds were good candidates for thermoplastic biopolymeric m ­ aterials38.
Bioplastic for food packaging might be ingested by human and animals. Additionally, most food packages are
designed for single use and very probably will be disposed to the environment. Therefore, materials to be used
for food packaging applications should be safe to consumers and the e­ nvironment39–41.
In this paper, PVA\GG composites have been prepared with three different compositions (50/50, 30/70,
and 70/30) which are denoted in this research as S1, S2 and S3, respectively. Films from the blends have been
prepared by solvent casting technique. The bioplastic films were characterized by FTIR, and the mechanical
properties were investigated by tensile tests. Plant extracts, namely, Guajava leaves (GL), and chickpeas (CP)
extracts have been prepared and characterized by Fourier transform infrared spectroscopy. The extracts were
incorporated individually to the most promising composition of the PVA/GG blends (S2 composition). Films
of PVA/GG and the plant extracts (S2/GL and S2/CP) have been investigated for their food packaging proper-
ties and utilization. To test the real performance of the prepared films as food packaging material, accelerated
weather testing and water wettability tests have been carried out. The bioplastic films have also been tested for
their cytotoxicity, oxygen, and water permeability. Life cycle assessment (LCA) and C ­ O2 emissions of the newly
prepared bioplastics have been studied.

Methodology
Chemicals and plant materials
Chemicals
PVA and GG were purchased from Sigma-Aldrich, USA. Glycerol was purchased from the united company for
chemicals, Cairo.

Plant materials
The plant materials collection and use in this research comply with the national and international guidelines and
legislation. GL were collected from national Guajava trees in Menofia. Chickpeas were commercial grade and
were purchased from national food store.

Bioplastic films preparation

PVA/GG bioplastic films preparation
PVA/GG films have been prepared by solvent casting ­technique25. PVA solution was prepared by dissolving 5 g
of PVA in 5 mL of distilled water and heated at 90 °C with stirring. GG, 2 g, was dissolved in 2 mL distilled water
and stirred at 70 °C until a clear solution was obtained. The two solutions were then mixed and stirred for 3–4 h
to obtain a completely homogeneous solution. Glycerol, 2 mL, was then added to the blend and stirred at room
temperature for 1 h. The mixture was then poured onto a clean and dry petri dish (200 × 20 mm) and dried in
an oven at 50 °C. This film with PVA/GG composition 70/30 is denoted as S3 in Table 1. The same procedures
have been repeated to prepare PVA/GG films with compositions 30/70 and 50/50 which were denoted as S2 and
S1, respectively. After drying, the films were peeled off and stored in a desiccator for further characterization.

PVA/GG/GL film preparation

Hot water (100 mL) at 85 °C has been added to 2 g of dry powdered guajava leaves and left to soak for 24 h. The
extract was then filtered and added to the PVA/GG blend of composition 2 (S2) in 5 wt% and stirred for ½ hr.
The blend PVA/GG/GL was poured onto clean and dry petri dish (200 × 20 mm) and dried in an oven at 50 °C
for 2 days. The film was then peeled off and stored in a desiccator for further characterization.

PVA/GG/CP film preparation

Ethanol–water mixture (1:1, 100 mL) was added to 2 g of finely powdered chickpea seeds and soaked for 24 h.
The extract was then filtered and added to the PVA/GG blend with composition 2 (S2) in 5 wt%. The PVA/GG/
CP blend was stirred for ½ hr, poured onto clean and dry petri dish (200 × 20) and dried in an oven at 50 °C for
2 days. The film was then peeled off and stored in a desiccator for further characterization.

Sample PVA (wt%) GG (wt%) Glycerol (mL)

S1 50 50 2
S2 30 70 2
S3 70 30 2

Table 1.  The different compositions of GG/PVA blends.

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Fourier transform infrared spectroscopy (FTIR)

The FTIR of all the films were recorded on a nicolet iS10 FT-IR spectrophotometer with a resolution of 4 ­cm-1
in the range 450 and 4000 ­cm-1.

Mechanical measurements
Tensile tests were performed by the ASTM D412 test strategy and were done at room temperature using a tensile
tester (Tinius Olsen-H10K). The crosshead speed was (10 mm/min). The initial gauge length of the specimen
was (80 mm) and the width of each sample was (2 mm). Thickness 0.130–0.150 mm.

Contact angle measurement

The contact angle measurements were performed using Attention Theta Optical Tensiometer (Biolin Scientific
Company, Finland).

Cytotoxicity studies for PVA/GG film, S2

Determination of S2 (PVA/GG, 30/70 wt%) cytotoxicity on Vero cells (kidney tissue) using MTT protocol has
been carried out following the procedures reported e­ lsewhere42 at a concentration 1000 ug/mL of S2.

Scanning electron microscopy (SEM) for the PVA/GG (S2) film

The surface morphology of the prepared PVA/GG film of composition 30/70 wt% (S2) was examined using FEI
Quanta FEG 250 high resolution scanning electron microscope (SEM).

Water vapor transmission rate (WVTR) measurements

The WVTR measurements for S2, S2/GL and S2/CP films were performed according to the method reported
­in43 using a samples of 1.5 cm diameter. The films under investigation were sealed on top of a glass bottle filled
with distilled water and placed in a desiccator at 90% relative humidity. The decrease in the weight was detected
using a microbalance at 24 h time intervals for 5 days. All experiments were carried out in duplicate. The WVTR
values were calculated from the slope of the regression line of the weight change/time graph using Eq. (1)43.
WVTR = � m/(A × t) (1)
Where (Δ m/t) is the slope of the weight change against time (g/day), A is the sample exposed surface area in ­m2.

Oxygen permeability
The oxygen permeabilities of samples S2, S2/GL and S2/CP were measured by the method described by Shi et al.44
The oxygen permeability of the samples under investigation has been calculated using Eq. (2)44.

(2)
   
OP oxygen permeability = Mf − Mi /(t x A)

where Mf is the final weight of the weighing bottle after equilibrated in the desiccator for 24 h at 90% humidity
and 25 °C, Mi is the initial weight of the weighing bottle and t is the equilibrated time (48 h) in seconds, and A
is the surface area of the exposed films.

Accelerated weather testing

The accelerated weathering of the bioplastic samples was conducted in an accelerated weathering tester Model
QUV/se (Q-LAB, Westlake, Ohio, USA). The weathering conditions were following the Cycle-C of the ASTM
D6164 standard. Fluorescent UV lamps (UV-A-340) with 0.76 W m-2 irradiance (wavelength 340 nm) were
used with cycles of 8 h UV irradiation at 50 °C, followed by 4 h dark at 50 °C under 100% condensing humidity.
These consecutive cycles were applied to the specimens attached to the test panels without any interruption. The
effects of accelerated weathering were investigated for 100 h exposure periods.

Life cycle assessment and carbon footprint for the prepared bioplastic
Life cycle assessment has been investigated using the model proposed by Benavides et al.45. The data for ­CO2
emissions for the different components in the bioplastic have been collected from different resources. The refer-
ences are provided in the references section for each value considered.

Results and discussion

preparation of PVA‑GG blend
PVA/GG blends were prepared by blending PVA (Fig. 1a) and GG (Fig. 1b) in three different compositions,
(50/50), (30/70) and (70/30) wt% denoted as S1, S2 and S3, respectively, Table 1.
It has been reported that, addition of glycerol by 20–70% to GG based bioplastic films enhanced the mechani-
cal ­properties23, therefore, glycerol, 25 wt%, was added to the three compositions as a plasticizer. Solvent casting
­technique26 has been used to prepare the bioplastic films, where the blends have been poured uniformly onto
dry and clean petri dishes and allowed to dry in an oven. After peeling, the films of the different compositions
have been kept in a desiccator for further characterization (Fig. 2).
The films of PVA/GG with the three compositions (S1, S2 and S3) have been characterized by FTIR. The
mechanical properties have also been investigated by tensile strength measurements.

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Figure 2.  The step wise preparation of GG/PVA bioplastic by solvent casting techniques.

Fourier transform infrared spectroscopy (FTIR) for the PVA/GG blends

The FTIR for the pure GG and PVA, in addition to the three blends have been investigated. The three composi-
tions of PVA/GG (S1, S2 and S3) gave almost the same bands in the IR spectra; therefore, the three compositions
have been represented by only one IR chart in Fig. 3. Figure 3 shows the IR for the pure GG, PVA and GG/PVA
blend. FTIR chart for pure GG showed the characteristics bands for GG. The large broad band at 3437 ­cm-1 is
corresponding to the -OH group vibration and the stretching of intermolecular hydrogen bonding. At 2891 ­cm-1,
is the ­CH3 and ­CH2 stretching vibrations. The band at 1627 ­cm-1 is probably corresponding to the C=O stretch-
ing while the band at 1031 ­cm-1 represents the glycosidic bond vibration. The band at 1412 ­cm-1 corresponds to
the CH b ­ ending46,47. The FTIR chart of the pure PVA is also shown in Fig. 3. The peak at 3469 ­cm-1 represents
the -OH group asymmetric stretching vibration resulting from the inter- and intramolecular hydrogen bonding
in pure PVA. The peak at 2941 ­cm-1 refers to the –CH stretching from the alkyl groups. The peak at 1726 ­cm-1

Figure 3.  The FTIR for pure GG (green line), Pure PVA (red line) and PVA/GG blend (blue line).

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corresponds to the –C=O stretching. The peak at 1450 ­cm-1 represents –CH bending while at 1100 ­cm-1 is due
to the -CO stretching of the –COH ­group48–50. After blending GG and PVA, a shift in the peak positions was
observed. The peak assigned to the -OH was broader and shifted to a lower wavelength (3298 ­cm-1) than in the
pure PVA and GG. This shift can be attributed to an increase in the intermolecular hydrogen bonding resulted
from the blending. The peak at 1726 ­cm-1 in PVA was also shifted to be at 1716 ­cm-1 which indicates an increase
in the hydrogen bonding. The peak at 1031 ­cm-1 in GG appeared at 1037 ­cm-1 in the mixture which indicates
successful ­blending23,49.

Guava extraction, characterization, and incorporation in the PVA/GG blend

It has been reported that Guava trees contribute largely to the C ­ O2 sequestration. They help in the reduction of
greenhouse gas emissions in the ­atmosphere51. A 10–15 years old Guava tree can remove from 12 to 13 kg/year
of ­CO2 from the ­atmosphere52. Therefore, in this work, we incorporated GL extract to the PVA/GG bioplastic
in order to reach a reduced net GHG emissions and consequently reduce the C ­ O2 footprint of the prepared bio-
plastic. Additionally, the GL extract would enhance the mechanical and the bioactive properties of the bioplastic
for food packaging a­ pplications30–34.
As the type of solvent used in plants extraction affects the amount of the bioactive material in the extract.
Water was the most commonly used solvent for extraction of the active compounds from GL because it is safe
and ­economic53. Therefore, in this paper, GL were extracted in hot distilled water. The leaves were dried before
extraction. This is due to the fact that the phenolic compounds and antioxidants are more concentrated in dried
plants than in fresh ­ones53,54. It has been reported that water extract of GL is rich in glycosides, saponins, tan-
nins, phenols, terpenoids and fl ­ avonoids31,33. The aqueous GL extract has been characterized by FTIR, Fig. 4a.
The broad band at 3450 ­cm refers to the -OH stretching of the phenolic compounds. The band at 2990 ­cm-1 is
-1

attributed to the -CH stretching of aliphatic hydrocarbons. At 1638 ­cm-1 is the –C=O stretching of the carbonyl
compounds. A band at 1070 ­cm-1 corresponds to the –CO of the flavonoids. The broad band at 610 ­cm-1 cor-
responds to the heterocyclic compounds in GL e­ xtract31,53.
In GL water extract, the –OH and .C=O functionalized compounds are predominant. The chemical structure
of the most abundant group present in GL extracts compounds is illustrated in Fig. 4c33,55. The GL extract has
been blended with PVA/GG (30/70) mixture in 5 wt%. The mechanical properties, water and oxygen perme-
abilities, water wettability and life cycle of the PVA/GG/GL (S2/GL) bioplastic have been investigated in fol-
lowing sections.

Chickpea extraction, characterization, and incorporation in PVA/GG bioplastic

Chickpeas are abundant seeds that are grown and consumed all over the world. The seeds are rich in protein and
many other nutrients. Protein based bioplastics have been reported as food ­packaging29. Protein-based materials
contain extra cross linking that may enhance the bioplastic properties and the hydrophobicity of the material.
Ethanolic extract of chickpea seeds has been reported to be rich in phenolic components and ­oligosaccharides56,57.

Figure 4.  (a) the FTIR of GL extract, (b) FTIR of CP extract, (c) the galloyl group present in GL extract active
­compounds50, and (d) an example of the hydroxy compounds presents in CP extract (genistein)61.

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In this paper, we targeted the phenolic and other hydroxy functionalized components in the plants extracts as
the -OH functionalized compounds are predicted to act as plasticizers-like materials. Therefore, an aqueous
ethanolic solution (50/50) has been used in the extraction of the active components in grounded chickpeas. The
FTIR chart for CP aqueous ethanolic extract has been shown in Fig. 4b. The broad peak at 3298 ­cm-1 corresponds
to the –OH and –NH stretching vibrations. The peak at 2933 ­cm-1 refers to the –CH and –CH2 stretching vibra-
tion. The peaks at 1650 and 1565 ­cm-1 are attributed to the amide groups (–CONH) of proteins. The sharp band
at 1034 ­cm-1 is referred to the –CH bending of the glycosidic ­bond29,58–60.
The chemical structure of an example of the active compounds present in GL extracts is illustrated in Fig. 4d61.
The CP extract has been blended with the PVA/GG mixture in 5 wt% and the blend is denoted as S2/CP in this
paper. The mechanical properties, water wettability, cytotoxicity, water and oxygen permeability and life cycle
of the S2/CP bioplastic have been investigated in following sections.

Mechanical properties
The mechanical properties of the prepared bioplastic have been investigated by measuring the tensile strength
for the films S1, S2 and S3 (Table 2) prepared by solvent casting technique. The tensile strength measurements
for S1, S2 and S3 have been shown in Fig. 5a–c, respectively. Figure 5d shows the tensile strength for S2, S2/CP
and S2/GL.

Sample (PVA/GG) TS (MPa) Strain (%)

S1 (50/50) 0.27 119
S2 (30/70) 0.20 6.19
S3 (70/30) 0.19 39
S2/GL 0.26 6.83
S2/CP 0.22 6.77

Table 2.  The tensile strength and elongation values of the prepared bioplastic films.

Figure 5.  The tensile strength measurement for (a) sample S1, (b) sample S2, (c) sample S3 and (d) S2, S2/CP
and S2/GL.

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From Table 2, S2 had a suitable tensile strength/strain values for a bioplastic material. The sample has the
least amount of PVA with a fossil origin, therefore, S2 blend was selected along with S2/plant extracts for further
studies. Tensile strength measurements for S2 after addition of the GL and CP extracts have been carried out
to investigate the enhancement in the bioplastic mechanical properties by the addition of plant extracts. It has
been reported that the addition of PVA to the GG enhanced the GG mechanical ­properties16. After the addition
of the GL and CP extracts, the mechanical properties of the bioplastic have been enhanced even further, Fig. 5d.
This enhancement can be attributed to the -OH functionalized components in the plant extracts which can act
as natural plasticizers. Additionally, the extract components with OH and NH groups can form hydrogen bonds
or physical crosslinking with the GG resulting in an improved mechanical property. Similar effects have been
observed by other research ­groups27,29,62. The more enhancement in the mechanical properties by addition of
GL extract than the CP extract can be attributed to the presence of small molecule phenolic compounds in GL
extract.

Water contact angles measurement

In food packaging, wettability of the plastic plays an important role in the food preservation process and during
transportation. A less water wet surfaces are desired in order to keep the packed food safe from m­ oisture63. The
water contact angle of the PVA/GG bioplastics have been measured, Fig. 6. For the PVA/GG (S2), The contact
angle was 87°. The addition of the CP and GL extracts to the PVA/GG blends increased the hydrophilicity of the
films. This can be attributed to the more polar components (hydroxyl group functionalities) presented by the
addition of the plant extracts. Additionally, film with GL extracts showed more water wetting than that with the
CP extracts. This may be attributed to the extraction solvents. GL has been extracted with pure water which may
result in the presence of more small polar compounds than those present in CP e­ xtract56,57,63.

Cytotoxicity studies
Cytotoxicity of the film S2 against the Vero normal cell line was assayed. Results showed that the cell viability of
­ ackaging63–65.
Vero cells was 99.67% (Fig. 7 and Table 3), which is considered very safe for S2 to be used in food p

Figure 6.  The water contact angle of the bioplastic films: (a) S2, (b) S2 with GL and (c) S2 with CP.

Figure 7.  (a) Control Vero cells and (b) Effect of PVA/GG (S2) on cell viability of Vero normal cell line at
concentration 1000 uL and incubation time 24 h.

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ID Optical density Mean optical density ± SE Viability %* Toxicity %

Vero 0.717 0.719 0.716 0.716 0.00208 100 0
S2 0.713 0.72 0.7136 0.713 0.00348 99.674115 0.35884

Table 3.  Viability and toxicity of S2 on Vero cell line. *Viability has been calculated using the formula
reported ­in42.

Scanning electron microscopy (SEM) for the PVA/GG film

Scanning electron microscopy for the surface of the sample showed a smooth surface with no cracks, Fig. 8.
Only few irregularities or voids have been observed on the surface of the film. The presence of voids might be
attributed to the degree of dispersion of the plasticizer which may result in poor crosslinking in few spots of the
bioplastic ­sample62. Similar observation in bioplastic films has been reported by Amritkumar et al.62 and other
research ­groups66,67.

Water vapor transmission rate (WVTR) measurement

The value of WVTR for the PVA/GG (S2) sample was very comparable to the values of the samples with plant
extracts (S2/GL and S2/CP), Table 4. Only slight increase in the WVTR values in the samples with the plant
extracts (S2/GL and S2/CP). This might be attributed to the small amount (5 wt%) of the plant extracts added
to S2 which caused slight increase in the hydrophilicity of the S2/GL and S2/CP. The plant extracts of GL and
CP are rich in hydroxy and phenolic compounds that might cause the slight increase in the hydrophilicity of the
samples S2/GL and S2/CP. In general, the bioplastic films, S2, S2/GL and S2/CP exhibited comparable WVTR
­ ackaging27,68–71.
values to other bioplastics frequently used for food p

Oxygen permeability (OP)

The oxygen permeability values for samples S2, S2/GL and S2/CP have been listed in Table 4. In general, the
OP values noticeably increased in the samples with the plant extracts (S2/GL and S2/CP). It has been reported
that the decrease in the hydrogen bonding can lead to a decrease in the oxygen ­permeability72. This observation
might explain the noticeable increase in the OP values for S2/GL and S2/CP compared to S2 as the addition of
the plants extract might reduce the uniformity of the hydrogen bonding within the samples.

Accelerated weathering test

In the accelerated weathering test, it was clear that all the bioplastic samples had undergone a visible change in
shape and color, Fig. 9. This indicates the biodegradability of the prepared bioplastics. Additionally, samples with
plant extract additives showed more cracks and had been torn. The increased tendency of bioplastics with plant
extracts to biodegradation can be attributed to the increased hydrophilic content (ability for hydrolysis) of the
samples due to the presence of polar compounds from the plant ­extracts73,74.

Figure 8.  Scanning electron microscopy for S2 bioplastic film.

sample WVTR (g/m2/d) OP (g/m2/s)

S2 5.73 18 × ­10−4
S2/GL 5.77 43 × ­10−4
S2/CP 5.74 67 × ­10−4

Table 4.  WVTR values (g/m2/d) and oxygen permeability values (g/m2/s) for samples S2, S2/GL and S2/CP.

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Figure 9.  Accelerated weathering test, (a) PVA/GG (S2), (b) S2 with GL extract and (c) S2 with CP extract.

Life cycle assessment and carbon footprint for the prepared bioplastic
Bioplastic is an environmentally friendly solution for the problematic accumulation of fossil-based plastics. The
biodegradability of bioplastic is considered an effective solution for plastic waste management. But at the same
time, biodegradability led to GHG emissions. Therefore, life cycle assessment (LCA) has become a prerequisite
to understand the environmental impact for any newly prepared bioplastics. A proposed LCA for the newly
prepared PVA/GG based bioplastic is illustrated in Fig. 1045,75. The LCA considered a comparison of a fossil
derived plastic, namely LDPE in addition to the newly prepared PVA/GG ­bioplastics45. The PVA/GG based
bioplastic composed of 30% PVA and 70% GG, therefore all the assessments and calculations consider the active
percentage of each component. The system boundaries in Fig. 10 covered all the materials (chemicals, water,
enzymes, etc.) and energy inputs for all stages including the conversion and production process of plastic and
possible end of life scenarios. The end-of-life proposed scenarios are landfilling for the LDPE and composting
for PVA/GG bioplastic.
To calculate C­ O2 emissions of the bioplastic, detailed GHG for each step and components have been con-
sidered and calculated. The GHG values for the bioplastic components and LDPE have been listed in Table 5.
The life cycle GHG emissions of LDPE and the PVA/GG based bioplastic have been illustrated in Fig. 11.
The carbon uptake for GG has been calculated based on the carbon content in its structure (based on its
molecular formula) while fossil-based LDPE, HDPE and PVA have no carbon uptake. The net GHG of the bio-
plastic is then equal to the GHG emissions minus the atmospheric ­CO2 taken or absorbed during the biomass
­cultivation45.
The value of plastic processing and production ­CO2 emission is equal for all plastic and bioplastic as reported
by Benavides et al.45 after conversion of energy input to C
­ O2 ­emission77. The biodegradation of PVA is associated
with the emission of 2 kg C ­ O2e/Kg ­PVA78. To the best of our knowledge, no exact values for the ­CO2 emissions
associated with GG degradation. Consequently, excluding the biodegradation value of GG, the total C ­ O2 emis-
sions for the PVA/GG (30/70) bioplastic is equal to 7.14 K ­ gCO2e/KG. This value is expected to decrease after
the addition of the plant extracts as guava and chickpeas trees are reported to have a potential in increasing
carbon ­sequestration51.

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Figure 10.  A proposed LCA for the prepared PVA/GG based bioplastic.

Component GHG emission ­(KgCO2e/Kg plastic) Source of information

PVA 5.5 Ref.75
LDPE 2.9 Ref.45
GG 7.8 Ref.76

Table 5.  GHG emissions values for the plastic and bioplastic components.

Figure 11.  The GHG emissions of PVA/GG based bioplastic in comparison with LDPE and HDPE.

Conclusion
In this work, three compositions of PVA/GG blends have been successfully prepared. The composition with
30/70 percent of PVA/GG showed promising mechanical properties as a bioplastic for food packaging applica-
tions. The addition of GL and CP extracts enhanced the mechanical properties even further due to the presence
of hydroxy functionalized compounds that can act as natural plasticizers. Scanning electron microscopy studies
for the surface of the sample S2 showed normal surface for plastics with few voids that might be attributed to
the degree of the dispersion of the plasticizer. Cytotoxicity assay for the new PVA/GG (S2, 30/70) showed that
the newly prepared bioplastic is very safe to be used for food packaging. In general, the WVTR test for the S2,
S/GL and S2/CP showed values that are comparable to previously reported values for other bioplastics martials
for food packaging applications. Oxygen permeability test showed that the addition of plant extracts leads to
an increase in the oxygen permeability for samples S2/GL and S2/CP compared to S2 (with PVA and GG only)
which might indicate that the compounds present in the added plant extract disturbed the hydrogen bonding
between the polymers leading to more gas permeation. The GL and CP bioactive phenolic compounds in their

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extracts increased the wettability and facilitated the degradation process under the optimum environmental
conditions. LCA has been carried out to facilitate carbon footprint calculations. The LCA and carbon footprint
calculations revealed that the use biopolymers derived from potential carbon sequestering biomass can largely
balance the carbon emissions from bioplastics. Consequently, bioplastic can be a safe solution for fossil based
plastic accumulation.

Data availability
All data generated or analyzed during this study are included in this article.

Received: 24 June 2023; Accepted: 12 December 2023

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Author contributions
All authors contributed equally to the article.

Funding
Open access funding provided by The Science, Technology & Innovation Funding Authority (STDF) in coopera-
tion with The Egyptian Knowledge Bank (EKB).

Competing interests
The authors declare no competing interests.

Additional information
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