Food Bioscience 41 (2021) 100977

Contents lists available at ScienceDirect

Food Bioscience
journal homepage: www.elsevier.com/locate/fbio

Development and characterization of corn starch-gelatin based edible films

incorporated with mango and pineapple for active packaging
Aliyath Susmitha a, b, Keerthi Sasikumar a, b, Devi Rajan a, Arun Padmakumar M a,
Kesavan Madhavan Nampoothiri a, b, *
a
Microbial Processes and Technology Division, CSIR- National Institute for Interdisciplinary Science and Technology (NIIST), Trivandrum, 695019, Kerala, India
b
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India

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

Keywords: The effect of corn starch-gelatin (CSG) films enriched (5–15% w/v) separately with mango (puree and peel) and
Active packaging pineapple pomace were characterized to analyse the functional properties for active packaging. The incorpo­
Antimicrobial ration of mango puree (MP), MP with peel (MPP) and pineapple pomace (PP) improved the physicochemical
Antioxidant
properties including moisture content, swelling index, thickness and opacity, while tensile strength and elon­
Biodegradable
gation at break was better for the control CSG films. The biological properties such as antioxidant, antimicrobial
Corn starch
Edible films activity and total phenolic content were increased with increase in concentration of all the films. The FTIR
Pineapple analysis showed a good interaction and blending of all the film constituents and the thermogravimetric analysis
(TGA) showed a closer resemblance of the films with that of the control. All films showed more than 50%
biodegradability after 15 days. These biodegradable films from these tropical fruits with enhanced properties
have the potential to be utilized for the formulation of edible food-packaging materials in the near future.

1. Introduction
According to the FAO reports, 14% of the food produced globally is
lost during the post-harvest production stage before reaching the retail
stage of the food system (Jenkins, Tucker, & Grim, 2016, p. 462). The
short shelf life and over ripening of fruits prior to their consumption is
one of the major problems faced by the society which may lead to their
wastage. These fruits can be converted into economically important
products such as edible packing materials which can play a decisive role
in improving the shelf life of the food. The packing material made of
fruits and its by-products are promising candidates which can make
remarkable changes in the food packaging industries by replacing the
excessive use of synthetic polymers such as plastics (Torres-león et al.,
2018). The environmental issues caused by these synthetic polymers
greatly demand the importance of bio-based active packaging systems.
Nowadays, the use of biodegradable active packaging systems
composed of biopolymers, provide a semi-permeable barrier that helps
to extend shelf life by reducing the migration of moisture, loss of solutes
from fruit respiration and oxidation reaction (Rangel-Marrón,
Montalvo-Paquini, Palou, & López-Malo, 2013). Among natural
hydrocolloids to form edible films, starch and gelatin are the primary
sources. Corn starch is a predominantly used starch polymer with high
biodegradability which makes it suitable for developing the film matrix
(Aisyah, Irwanda, Haryani, & Safriani, 2018; Ghanbarzadeh, Almasi, &
Entezami, 2011). Gelatin is a digestible protein, containing nearly all the
essential amino acids, except tryptophan (Fakhouri, Martelli, Caon,
Velasco, & Mei, 2015). The films made up of starch-gelatin blends have
good film-forming properties (Rosseto, Krein, Balbé, & Dettmer, 2019)
and enhances with the addition of fruit by-products. These packaging
systems improves the shelf life of corn starch/gelatin films (CSG) by
preventing food spoilage and also provides a positive effect on food
quality, food safety and aroma of food (Adilah, Jamilah, Noranizan, &
Hanani, 2018; Otoni et al., 2017).
Mango and pineapple are most popular tropical fruits well known for
its distinctive flavour, taste and nutritional value (Farid Hossain, 2015;
Melo et al., 2019). These fruits are highly perishable and consumed both
fresh and processed. During their processing, large amounts of
by-products consisting mainly of peel, seed, core and pomace were
generated, which contains several health promoting compounds (Selani
et al., 2016). Mangoes produce 13.5% of seeds, 11% of peels, 17.94%

* Corresponding author. Microbial Processes and Technology Division, CSIR- National Institute for Interdisciplinary Science and Technology (NIIST), Trivandrum,
695019, Kerala, India.
E-mail address: madhavan@niist.res.in (K.M. Nampoothiri).

https://doi.org/10.1016/j.fbio.2021.100977
Received 21 December 2020; Received in revised form 3 March 2021; Accepted 9 March 2021
Available online 13 March 2021
2212-4292/© 2021 Elsevier Ltd. All rights reserved.
A. Susmitha et al.
unusable pulp, and 57.56% of final products and pineapples produce
9.12% of core, 13.48% of peels, 14.49% of pulp, 14.87% of top, and
48.04% of finished products (Varzakas, Zakynthinos, & Verpoort, 2016).
Mangoes are climatric fruits which will continue to ripen after har­
vesting. During fruit processing, only the selected high-quality mangoes
go for further processing while the damaged or decayed mangoes are not
utilized. These underutilized fruits when disposed into the environment
cause various waste management issues. These risks can be minimized
by converting underutilized fruits into edible packaging films on whole
and minimally processed foods to maintain their quality and shelf-life.
The mango pulp and peel, and pineapple pomace are rich source of
dietary fibre and plays a significant role in enhancing the nutritional
quality of the food (Nguyen, Nguyen, & Savage, 2019; Selani et al.,
2014). The mango peels and pineapple pomaces are rich in antioxidants
which could improve the food stability, storage and texture by pro­
tecting it from oxidation and microbial attack. These natural properties
present in the mangoes lower the addition of synthetic compounds and
also reduces their release into the food which may cause some adverse
health problems (Melo et al., 2019). High levels of pectin present in the
mango can act as a natural jellying or thickening agent (Nguyen et al.,
2019). Besides the natural ingredients, there is a high requirement for
the addition of plasticizers that disrupts the hydrogen bonding between
the polymer chains, increases the flexibility and enhances the perme­
ability of gas and water vapour (Bourtoom, 2008). The common plas­
ticizers used include polyethylene glycol, sorbitol and glycerol. The type
and concentration of each plasticizers can have effect on the mechanical
properties of the film (Afifah, Ratnawati, & Darmajana, 2019). The
addition of sorbitol makes the film stronger and elastic than the standard
(Rahmawati, Arief, & Satyantini, 2019).
The purpose of the study was to develop edible and eco-friendly
biodegradable packaging system based on corn starch/gelatin (CSG)
blending incorporated with different concentration of mango puree
(MP), mango puree with peel (MPP) and pineapple pomace (PP), using
sorbitol as the plasticizer and also to evaluate its physicochemical,
mechanical, and biological properties such as total phenolic content,
antioxidant and antibacterial activity. In order to increase the antioxi­
dant and antibacterial value of the produced films, mango peel was
added in combination with mango pulp. This is the first report showing
the combination of mango peel with puree for the synthesis of edible
films.
2. Materials and methods
2.1. Materials
Mango, pineapple and corn starch were purchased from a local shop
(Trivandrum, Kerala, India). 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and
D-sorbitol were purchased from Sigma-Aldrich (Germany). Folin-
Ciocalteu reagent was acquired from SRL (India). All other chemicals
were procured from Himedia and Merck with analytical grade.
2.2. Preparation of films
Three different polysaccharide-gelatin based films, mango puree
(MP), mango puree with peel (MPP) and pineapple pomace (PP) films
were prepared. For the preparation of MP, ripened mangoes were first
washed, peeled, chopped using a blender to form a thick paste. For
making MPP, the sliced mangoes along with peel were homogenized.
Similarly, for extracting PP, fresh pineapples were washed, de-headed,
removed the peel and collected left over pomaces after juice extrac­
tion. To prepare single component film-forming solution (FFS), 0, 5, 10
and 15% (w/v) concentration of each MP, MPP and PP (around 13–19%
of dry matter in 100 g) were dissolved in a constantly stirred mixture of
distilled water (100 mL) along with 2% (w/v) corn starch and dissolved
at 75 ◦ C for 15 min. Then, 3% (w/v) of gelatin was added and heated at
75 ◦ C for 20 min. As plasticizer, 3% (w/v) of D-sorbitol was added to the
2
Food Bioscience 41 (2021) 100977
solution mixture with continuous stirring for another 35 min (the per­
centage of plasticizer is approximately corresponds to around 42.85% of
the polymer dry weight in pomace films and 40.98% of the polymer dry
weight in puree films). The films were prepared following basically the
study of Tulamandi et al. (2016) with slight modifications that we
optimized. In short, after cooling for 10 min, 100 mL of each film
forming solution (FFS) were filtered using a stainless-steel strainer with
mesh (0.2 mm). Thereafter, the whole 100 mL of the FFS were casted in
an open polycarbonate Petri dish of dimension (196 mm diameter x 19
mm height). Petri dishes were placed in a BOD Incubator (Lab line,
India), and the FFS was allowed to evaporate. The films were dried at 28
± 1 ◦ C for 28 h with a relative humidity of 54%. The films were then
gently peeled off and stored in air tight packets for further analysis. The
overall process flow and the films made were shown in Fig. 1.
2.3. Characterization of edible films
2.3.1. Moisture content
Moisture content of the films were measured according to the
method described by Torres-león et al. (2018). Film samples (2 cm × 2
cm) were weighed (W0) initially and dried in an oven at 105 ± 2 ◦ C for
24 h. Then, the dry weight (W1) of the films were measured and the
moisture content was determined using equation (1):
(W0 − W1 )
MC(%) = × 100 (1)
W0
where, MC % is the percentage of moisture content, W0 is the initial
weight and W1 is the dry weight of the film. All the tests were carried out
in triplicates.
2.3.2. Water solubility
The water solubility of edible films were measured according to
Sucheta, Rai, Chaturvedi, and Yadav (2019) with slight modifications.
The films were dried at 105 ± 2 ◦ C for 24 h and then were immersed in
tubes containing 20 mL of distilled water and incubated at 25 ◦ C, 150
rpm for 24 h. The insoluble fractions of the films were filtered using filter
papers and subjected to further drying in hot air oven at 105 ± 2 ◦ C for
another 24 h. The dried insoluble film fractions were further weighed
(W2) and the film solubility was calculated using equation (2):
(W1 − W2 )
WS(%) = × 100 (2)
W1
where, WS % is the percentage of water solubility, W1 is the dry weight
and W2 is the dry weight of the insoluble fraction of the film. All the tests
were carried out in triplicates.
2.3.3. Swelling index
The swelling index of the films were determined according to Cao,
Fu, and He (2007) with slight modifications. The films were cut into a
small pieces (2 cm × 2 cm), dried at 105 ± 2 ◦ C for 24 h and weighed
(W0) initially. The dried films were further immersed in 15 mL distilled
water for 2 min at
SI(%) = (W1 − W0 ) × 100 (3)
25 C. Later, the swelled samples were wiped with filter paper to
◦
remove excess liquid and weighed (W1). The amount of absorbed water
was calculated using equation (3):
W0
where, SI % is the percentage of swelling index and W0 and W1 are the
weights of dried and the wet samples. Each measurement was taken in
triplicates.
A. Susmitha et al.
Fig. 1. Schematic diagram showing the process development and final films made from mango (puree and peel) and pineapple pomace.
2.3.4. Film thickness
The thickness of the films (mm) was measured using a Mitutoyo
micrometer (Tokyo, Japan) with a sensitivity of 0.01 mm. Each film
sample was measured randomly at five different locations and the mean
value was reported.
2.3.5. Mechanical properties
The tensile strength (TS) and elongation at break (EAB) of the edible
films were measured by Universal Testing Machine (Hounsfield Ltd.,
Instron, UK). The films were cut into strips with 20 mm wide and 120
mm long. The 5 kN load cell was used to determine the mechanical
properties of samples (Lee, Lee, & Han, 2020). The initial grip distance
was 50 mm and the crosshead speed was 1 mm/min. The reported values
were the average of four measurements of each film.
2.3.6. Opacity measurement
According to Nouraddini, Esmaiili, and Mohtarami (2018) the films
were cut into rectangle pieces (1 cm × 4 cm) and placed directly in a
quartz spectrophotometer cell. An empty test cell was used as the
reference. The absorbance was measured at 600 nm using UV–Vis
spectrophotometer and the opacity of the film was calculated using
equation (4):
Abs600
O=
T (As)
where, O is the opacity, Abs600 was the absorbance at 600 nm and T is
the film thickness (mm). The analysis was conducted three times for
each type of the film and the mean value was reported.
2.3.7. Fourier transform infrared spectroscopy
Fourier transform infrared spectroscopy (FTIR) of different film
samples were analyzed using an IRTracer-100 (Shimadzu Corp., Kyoto,
Japan) FTIR spectrometer with an attenuated total reflection (ATR)
accessory. The chemical structure analysis of edible films was obtained
from an average of 45 scans per minute at a resolution of 4 cm− 1 over a
spectral range of 400 cm− 1 and 4000 cm− 1. Data analysis of each edible
film was done with Origin 8.5 program.
2.3.8. Thermogravimetric analysis (TGA)
The thermal stabilities of the control and blended films were inves­
tigated using thermogravimetric analyzer STA-6000 (PerkinElmer,
Massachusetts, USA). 10–15 mg of each film samples were used for the
analysis. Film samples were heated at a constant temperature rate of
10 ◦ C/min from 50 to 500 ◦ C under 30 mL/min of nitrogen atmosphere.
Food Bioscience 41 (2021) 100977
2.3.9. Total phenol content
The total phenolic content was determined using Folin-Ciocalteu
reagent according to Moghadam, Salami, Mohammadian, Khodadadi,
and Emam-Djomeh (2020). A standard curve was plotted using gallic
acid at different concentration (0–100 μg/mL). The results were
expressed as the milligram of gallic acid equivalent per unit gram weight
of the film (mg GAE/g film).
2.3.10. Evaluation of antioxidant properties
2.3.10.1. DPPH radical scavenging activity. The 2-diphenyl-1-picrylhy­
drazyl (DPPH) free radical scavenging assay described by Moghadam
et al. (2020) was applied to evaluate the antioxidant properties of the
edible films. The 900 mg of the film samples were added to each conical
flask containing 18 mL methanol (covered with aluminium foil) and
incubated at 25 ◦ C for 3 h with a gentle stirring (150 rpm). After the
incubation, the supernatant was analyzed for DPPH activity. The assay
was performed by adding 2 mL of 0.06 mM DPPH to 1 mL of the su­
pernatant. The methanol solution without film was taken as the control
for this experiment. The mixture was then vortexed and incubated fo r
30 min in dark at room temperature. The experiments were done in
triplicates. The antioxidant activity (AA) was measured using equation
(5):
(4)
AA(%) = 1 − × 100 (5)
(Ac)
Here, AS is the absorbance of the sample solution and AC is the
absorbance of the methanolic DPPH solution (control) at 517 nm. The
antioxidant activity (AA) was expressed as % DPPH radical scavenging
activity/100 mg of film.
2.3.10.2. Ferric (Fe3+) reducing power. Ferric reducing power assay was
determined according to Jridi et al. (2019). For reducing power assay,
0.5 mL of each film extract (as mentioned in section 2.3.10.1) was mixed
with 1.25 mL potassium ferricyanide (1% w/v) and 1.25 mL of phos­
phate buffer (0.2 M, pH 6.6) and incubated at 50 ◦ C for 20 min. The
obtained solution was mixed with 1.25 mL of 10% TCA (v/v) and
centrifuged at 4000 rpm for 10 min. Finally, to 1.25 mL distilled water
containing 0.1% FeCl3 (w/v), an equal volume of the supernatant was
added. The experiments were done in triplicates and the absorbance was
recorded at 700 nm.
2.3.11. Antibacterial activity
The antibacterial activities of the edible films were studied according
to the zone of inhibition in the disc diffusion method following the
3
A. Susmitha et al. Food Bioscience 41 (2021) 100977

procedure of Moghadam et al. (2020). Petri dishes containing
Muller-Hinton agar (MHA) were prepared. Overnight incubated cultures
of Pseudomonas aeruginosa, Salmonella typhi and Mycobacterium smeg­
matis in Luria Bertani (LB) broth were seeded on to MHA plates by swab.
Films were cut into pieces (1 cm × 1 cm) and exposed to UV for 20 min.
The UV treated films were placed onto the inoculated MHA plates using
sterile forceps and the plates were incubated at 37 ◦ C for 24 h. After
incubation, the diameter of the zone of inhibition (mm) was measured.
2.3.12. Biodegradability
Composting test was conducted according to Nouraddini et al.
(2018). Film samples (1 cm × 2 cm) were cut, weighed and placed on an
iron wired gauze and buried at 2 cm depth in an aluminium tray con­
taining natural soil at room temperature. Everyday water was sprayed to
the soil. After 15 days, the film samples were taken out for photograph.
The degradation of the films was evaluated in terms of percentage of
weight loss using equation (6) (Sekhar et al., 2016):
(W0 − W1 )
WL(%) = × 100
W0
Aelma Husna, Nurul Syahida, Nor Khaizura, & Jamilah, 2018).
3.2. Water solubility
The solubility of corn starch-gelatin films (CSG) incorporated with
different concentration of MP, MPP and PP are shown in Table 1. The WS
of the control film was found to be highest (64.07%) due to its hygro­
scopic nature and its solubility significantly (p < 0.05) decreased with
the incorporation of MP, MPP and PP at different concentration. The
high solubility in control film may be due to hygroscopic nature of starch
and gelatin which disintegrates in water rapidly (Ramos, Valdés,
Beltrán, & Garrigós, 2016; Shih, 1996). The incorporation of MP, MPP
and PP in the formulations promoted a reduction in the water solubility.
The hydrophilic nature of the fibers in the fruits may interact with hy­
droxyl groups of both starch and gelatin and reduce the water solubility
of the films (Dantas et al., 2014). The edible films with low water sol­
ubility exhibits high water resistant which is an essential functional
parameter for the packing of intermediate- or high-moisture foods from
(6) spoilage (Galus & Kadzi, 2016).

where, WL % is the percentage weight loss of the film, W0 is the initial
weight of the film and W1 is the weight of the film after degradation.
2.3.13. Statistical analysis
All the results were analyzed using one-way analysis of variance
(ANOVA) to evaluate the statistical significance of the data at p < 0.05
and presented as mean ± standard deviation. The SPSS 22 statistical
software (IBM Corporation) was used for data analysis. The mean
comparisons were conducted using Tukey’s test to examine if the dif­
ferences between each samples were significant (α = 0.05).
3. Results and discussion
3.1. Moisture content
Moisture content of different film samples are presented in Table 1.
In comparison to the CSG (control), the moisture content (MC) of MP,
MPP and PP films were found to be significantly (p < 0.05) high. The
addition of MP, MPP and PP into the film-forming solution slightly
increased the moisture content of the films which could be due to the
presence of hydrophilic components (proteins, carbohydrates and fiber)
within the fruits which enables them to interact with water molecules,
thus increasing the MC of the films (Nouraddini et al., 2018). The MC for
PP film was comparatively less than MP and MPP films, which may be
due to the low fiber content in pineapples than in mangoes. The films
with high moisture content are found to be more stretchable and flex­
3.3. Swelling index
Swelling index of edible films indicates their biodegradation and
maintenance of quality and storage of food products (Cerqueira, Souza,
Teixeira, & Vicente, 2012). The results confirmed that the swelling ca­
pacity of CSG films was the lowest (24.58%), while with the addition of
MP, MPP and PP at different concentrations, swelling index values
significantly (p < 0.05) increased (Table 2). The swelling capacity of the
MP, MPP and PP films were found to be higher than that of the control
which indicates a greater hydration capacity of the films, which is an
important characteristic in food packaging. The swelling index of the
film was dependent on the incorporation fruit pulp and it’s by products
and the polysaccharide used in the formulation. These results were
similar to those reported for the film made from low methoxy pectin
(LMP) and jambolan juice which showed a higher swelling index (Hulda,
Bianca, Wiliene, Daniel, & Flávio, 2020).
Table 2
Thickness, tensile strength (TS), elongation at break (EAB) and opacity of CSG
films incorporated with various concentrations of mango (puree and peel) and
pineapple pomace.
Samples Thickness TS (MPa) EAB (%) Opacity (A600/
(mm) mm)

ible, which could be used in various food application (Nur Hanani, CSG 0.22 ± 0.00e 0.50 ± 65.60 ± 8.36a 1.08 ± 0.14e
0.04abcd
5% MP 0.24 ± 0.00de 0.51 ± 46.73 ± 1.42 ± 0.04d
0.08abc 7.76bc
Table 1
10% MP 0.25 ± 0.01cd 0.56 ± 0.10ab 61.00 ± 1.69 ± 0.04c
Moisture content (MC), water solubility (WS) and swelling index (SI) of CSG 6.41ab
films incorporated with various concentrations of mango (puree and peel) and 15% MP 0.25 ± 0.00cd 0.34 ± 0.00cd 37.73 ± 2.39 ± 0.00a
pineapple pomace. 3.45cd
5% MPP 0.27 ± 0.02bc 0.32 ± 0.02d 35.40 ± 1.69 ± 0.14c
Samples MC (%) WS (%) SI (%)
2.42cd
CSG 13.7 ± 0.44d 64.07 ± 3.68a 24.58 ± 3.44f 10% 0.28 ± 0.00ab 0.39 ± 34.00 ± 1.40 ± 0.01d
5% MP 17.02 ± 1.12cd 35.4 ± 3.80bc 63.65 ± 2.09cde MPP 0.03bcd 5.29cde
10% MP 19.76 ± 0.51abc 49.78 ± 1.75ab 73.52 ± 3.70c 15% 0.30 ± 0.00a 0.59 ± 0.13a 42.47 ± 1.85 ± 0.01bc
15% MP 22.81 ± 1.78a 49.89 ± 8.39ab 73.65 ± 3.62c MPP 8.03cd
5% MPP 18.30 ± 1.4bc 30.50 ± 3.37c 74.24 ± 1.48bc 5% PP 0.25 ± 0.00cd 0.36 ± 0.03cd 30.26 ± 1.06 ± 0.00e
10% MPP 17.05 ± 0.67c 41.83 ± 4.45bc 54.82 ± 3.39e 3.98de
15% MPP 21.34 ± 1.10ab 26.82 ± 9.44c 66.44 ± 1.19cd 10% PP 0.22 ± 0.00e 0.42 ± 30.80 ± 1.79 ± 0.01bc
5% PP 16.44 ± 2.71cd 45.24 ± 7.55b 88.93 ± 6.70a 0.03abcd 2.84cde
10% PP 14.10 ± 0.61d 52.28 ± 11.23ab 84.88 ± 6.64ab 15% PP 0.27 ± 0.00bc 0.43 ± 19.03 ± 0.86e 1.90 ± 0.02b
15% PP 16.88 ± 1.80cd 50.53 ± 0.88ab 58.80 ± 1.21de 0.00abcd

a-f Means with different letters within a column are significantly different a-e Means with different letters within a column are significantly different
(ANOVA/Turkey’s test, p < 0.05). (ANOVA/Turkey’s test, p < 0.05).
MP: Mango puree, MPP: Mango puree with peel and PP: Pineapple pomace. MP: Mango puree, MPP: Mango puree with peel and PP: Pineapple pomace.

4
A. Susmitha et al.
3.4. Thickness
The thickness of the films is presented in Table 2. It was observed
that the thickness values for CSG (control), MP, MPP and PP films
ranged from 0.22 to 0.30 mm. The thickness of the films showed a sig­
nificant increase (p < 0.05) with the addition of MP, MPP and PP at
higher concentration. In agreement, Adilah et al. (2018) reported that
thickness of the films increased with the addition of high concentration
of mango peel extract (MPE) to gelatin films. Our results are also similar
to those presented by Rodríguez, Sibaja, Espitia, and Otoni (2020), who
indicated that the thickness of the papaya edible films was from 0.13 to
0.32 mm, and for carrot edible films the thickness ranges from 0.099 to
0.238 mm reported by Wang, Liu, Wei, and Ma (2011). The increase in
thickness of the films, may be related to some conformational changes in
CSG chains occurred by the addition of MP, MPP and PP at 15%. This
was in agreement with Tongnuanchan, Benjakul, and Prodpran (2014),
whereby the addition of extract compounds disrupted the ordered
alignment of gelatin film matrix. According to the literature reports, the
thickness of the films produced during research will be usually less than
0.3 mm (Jancikova, Dordevic, Jamroz, Behalova, & Tremlova, 2020).
Therefore, it is essential to have films with low thickness value, since the
packaged food product can be eaten along with the edible films.
3.5. Mechanical properties
The tensile strength (TS) and elongation at break (EAB) plays a sig­
nificant role in maintaining the structural integrity of the films. The
mechanical properties for different types of CSG films produced during
the study are tabulated in Table 2. The addition of MP, MPP and PP in
films caused significant (p < 0.05) differences in TS and EAB compared
to CSG films. Moreover, the addition of 5% and 10% MP, and 15% MPP
caused an increase in strength in comparison with CSG. However, the
EAB of the films decreased when MP, MPP and PP was added at different
concentration. The similar results was observed by Nouraddini et al.
(2018), where a decrease in TS and EAB of corn starch films were
observed after the addition of eggplant flour (EF). The presence of
different antioxidant agents have a pronounced impact on mechanical
properties of the film (Gallego, Gordon, Segovia, & Almajano Pablos,
2016). In fact, the decrease in mechanical properties depends on the
level of MP, MPP and PP incorporated. A possible reason is that the free
sugars present naturally in mangoes as glucose, fructose and sucrose
acted as plasticizers. The addition of plasticizer (sorbitol) to the film
base have increased the final concentration, resulting in excessive
interaction between the film network and the plasticizer, thereby
reducing film flexibility (Ioannis Arvanitoyannis, Psomiadou, &
Nakayama, 1996).
3.6. Opacity
The opacity plays an important role to control the light incidence
over the food. As shown in Table 2, the incorporation of MP, MPP and PP
additives significantly (p < 0.05) increased the opacity of CSG films,
except 5% PP showing an opacity value similar to CSG films. The slight
increase in values may be due to the addition of polyphenols, anthocy­
anins and carotenoids present in mango and pineapple. This is in
agreement with Nur Hanani et al. (2018), where the increase the opacity
in the mango peel is due to the presence of carotenoids and anthocyanin
contents, which absorbs lights at these wavelength. Moreover, Li, Miao,
Wu, Chen, and Zhang (2014) reported that the –OH group in poly­
phenolic compounds reduces the transmission of light. Although, the
films that are too opaque reduces its exposure to UV–Vis light and
controls oxidative deterioration which may exhibit discolouration and
off-flavours in food (Yuan, Lv, Yang, Chen, & Sun, 2015). Therefore, the
films with low opacity were preferred for commercialization, since it has
a direct influence over product and consumer acceptance (Nur Hanani
et al., 2018; Ojagh, Rezaei, Razavi, & Hosseini, 2010).
5
Food Bioscience 41 (2021) 100977
3.7. FTIR
The FTIR spectra in Fig. 2 a, b and c indicates that the addition of MP,
MPP and PP did not change the molecular interaction of the formed
films. However, the absorbance pattern were found to be identical to
that of the control due to their similar elemental composition. The broad
overlapping peaks of the films at 850.61 cm− 1 and 923.90 cm− 1 shows
the presence of corn starch which attributed to the vibration of glucose
pyranose and C–O vibrations stretching of glucose units respectively
(Ibrahim, Sapuan, Zainudin, & Zuhri, 2019). Likewise, the occurrence of
C–OH group in the films structure showed a peak at 1076.29 cm− 1
whereas the coupling of C–C and C–O bond stretching appears at
1149.57 cm− 1 which is a characteristic peak of the polysaccharide
molecules (Pourfarzad, Ahmadian, & Habibi-Najafi, 2018). A peak was
observed at 1367.53 cm− 1, which occurred due the blending mode of
CH2. A sharp peak at 2924.08 cm− 1 has occurred due to C–H vibrational
stretching. While, an extreme band at 3265.48 cm− 1 was observed due
to O–H group vibrational stretching (Ibrahim et al., 2019). The sorbitol
spectra has maximum absorption within a range of 850–1450 cm− 1
indicating C–OH stretching vibration and O–H bond vibration within
this region (Quinquenet, Ollivon, Grabielle-Madelmont, & Serpelloni,
1988). The spectrum of gelatin exhibits a characteristic peak in the re­
gion of 3000–3600 cm− 1 which indicates the presence of O–H group, at
1579.70 cm− 1 for amino band and 1656.85 cm− 1 for the carbonyl peak
of the amide group. The highest intensity of vibration in the range from
3100 to 3500 cm− 1 indicates N–H stretching vibration, which may be
due to the overlapping of O–H bond (Parvez et al., 2012).
3.8. Thermogravimetric analysis
Thermogravimetric analysis is a continuous processes used to un­
derstand the correlation between the sample weight changes in accor­
dance with increase in temperature (Parvez et al., 2012). The results of
the thermal decomposition behaviour of MP, MPP and PP films with
respect to CSG films are represented in Fig. 3. The control, MP, MPP and
PP films were analyzed using weight loss measurements. The CSG
(control) and MP (5% and 10%) showed an initial weight loss at 150 ◦ C
(Fig. 3a) while, the thermal stability was improved at high concentration
of MP (15%). The high thermal stability of MP at 15% was due to
increasing dissociation energy owing to numerous interactions occur­
ring between the polymer blend constituents (I Arvanitoyannis, Pso­
miadou, Nakayama, Aibab, & Yamamotob, 1997). The TGA
thermograms for MPP films showed that the CSG films undergo an initial
weight loss earlier than that of MPP films at different concentration
(Fig. 3b). Whereas as the initial weight for PP films was similar to CSG
films (Fig. 3c). All the films were stable up to 200 ◦ C and showed a
thermal decomposition at range of 250–280 ◦ C.
3.9. Total phenolic content
The Folin-Ciocalteau assay is a useful tool to measure the total
phenolic compounds present in the films. Based on the results summa­
rized in Table 3, TPC values of MP, MPP and PP significantly (p < 0.05)
increased when compared to pure CSG films. This result corresponds to
study reported by Adilah et al. (2018), who observed an increase in TPC
value with the enrichment of mango peel extract. Moghadam et al.
(2020) also reported the similar results, in which they observed the
mung bean protein films incorporated with pomegranate peel extract
showed a higher TPC values compared to control films. Among all the
films, the highest TPC values are observed for 10% MPP, ranging from
13.17 μg GAE/g to 27.34 μg GAE/g. The TPC values of MP was signifi­
cantly decreased when compared to MPP and PP, this may be related to
the presence of high phenolic compounds found in mango peels and
pineapple pomace. The total phenolic content of ripe mango pulp of
different mangoes ranged between 33.16 and 120.14 mg GAE/100 g. In
contrast to pulp, mango peel contained higher total phenolic ranging
A. Susmitha et al. Food Bioscience 41 (2021) 100977

Fig. 2. FTIR spectrum of CSG films with different concentration a) Mango puree (MP), b) Mango puree with peel (MPP) and c) Pineapple pomace (PP).

Fig. 3. Comparison of TGA graphs of CSG films with various concentration of a) Mango puree (MP) b) Mango puree with peel (MPP) and c) Pineapple pomace (PP).

enhancing the nutritional and aesthetic quality of the food (Ecole & Sup,
Table 3
1996). Antioxidant properties of MP, MPP and PP films were measured
Total phenolic content, antioxidants (DPPH and reducing power) of CSG films
by DPPH free radical scavenging and Fe3+ reducing power assay
incorporated with various concentrations of mango (puree and peel) and pine­
(Table 3). The results showed that DPPH scavenging activity could be
apple pomace.
observed in all CSG films, but the effect of the films was found to be more
Samples TPC (μg GAE/g DPPH scavenging Fe3+ Reducing Power (Abs
significant (p < 0.05) when higher concentration of fruit additives were
film) (%) 600 nm)
added. The incorporation of 15% of MP, MPP and PP significantly (p <
CSG 13.17 ± 0.004d 12.59 ± 0.05i 0.08 ± 0.006f 0.05) resulted a higher DPPH value, compared to CSG control films.
5% MP 14.55 ± 0.002d 13.42 ± 0.22i 0.10 ± 0.007def
10% MP 15.24 ± 0.002cd 18.92 ± 0.24h 0.11 ± 0.008de
Generally from our findings, mango pulp with peel showed an increased
15% MP 16.62 ± 0.002cd 26.90 ± 0.42f 0.15 ± 0.002c antioxidant activity (82.83%) at higher concentration of 15% when
5% MPP 27.66 ± 0.001a 46.00 ± 0.26c 0.25 ± 0.006b compared to mango pulp (26.90%) alone. This may be due to the
10% 28.34 ± 0.003a 66.20 ± 0.44b 0.28 ± 0.001a considerable amount of biologically active compounds in mango peels
MPP
like carotenoid, anthocyanins, polyphenols and flavonoids (Torres-león
15% 23.51 ± 0.006ab 82.87 ± 0.71a 0.24 ± 0.006b
MPP et al., 2018). A study by Nur Hanani et al. (2018) showed that the
5% PP 14.21 ± 0.005d 24.54 ± 0.50g 0.09 ± 0.001ef addition of pomegranate, papaya and jackfruit peel greatly increased the
10% PP 19.38 ± 0.001bc 32.69 ± 0.23e 0.12 ± 0.008d DPPH value, as the presence of biologically active compounds in the
15% PP 17.66 ± 0.003cd 44.11 ± 0.30d 0.11 ± 0.002de films caused it to accept electrons or hydrogen atoms. The control films
a-i Means with different letters within a column are significantly different ( also showed DPPH scavenging activity due to the presence of amino
ANOVA/. acids in gelatin (Nur Hanani et al., 2018).
Turkey’s test, p < 0.05). Ferric reducing power assay evaluates the samples ability to reduce
MP: Mango puree, MPP: Mango puree with peel and PP: Pineapple pomace. ferric to ferrous ion (Benzie & Strain, 1999). Table 3 shows that the
Ferric reducing power values of MP, MPP, and PP significantly (p <
between 278.75 and 379.17 mg GAE/100 g (Muralidhara, Veena, 0.05) increased, compared to CSG films. As in the DPPH assay, control
Bhattacherjee, & Rajan, 2019) respectively, and pineapple pomace also showed a slight ability to reduce ferric ions. In this assay, MPP films
contains around 642 mg GAE/100 g (Montalvo-González et al., 2018). had the highest ability to reduce ferric to ferrous ion, followed by MP
The phenolic content of the film was found to be lower when comparing films, while PP films showed the lowest ability at an absorbance of 700
with TPC of mango pulp, mango peel and pineapple pomace, this could nm. This increase in MPP film could be associated with higher TPC
be due to the degradation of phenolic compounds at high temperature content in films improved after enriching with mango peel. Therefore,
during film formation. the films incorporated with these fruit extract are found to be effective in
food packaging for shelf life extension.
3.10. Antioxidant properties
3.11. Antibacterial activity
Antioxidant is a vital aspect to inhibit the free radicals. Moreover,
films enriched with natural antioxidants are considered to be effective in The antimicrobial activity of the edible films against Gram-positive

6
A. Susmitha et al.
(Mycobacterium smegmatis) and Gram-negative bacteria (Salmonella typhi
and Pseudomonas aeruginosa) were evaluated by determining the inhi­
bition zones (mm) on MHA medium and results reported in Table 4. The
incorporation of MP, MPP and PP films significantly (p < 0.05) showed a
wider zone of inhibition against the pathogens with the increase in
concentration. MPP films with highest concentration (15%) was more
effective and showed a wider zone of inhibition of 33.66 ± 0.58 mm
than MP films (15%) which gave a halo of 26.33 ± 1.25 mm against
P. aeruginosa after 24 h of incubation (Fig. 4), whereas the control films
did not show any antibacterial activity against all the three tested mi­
croorganisms. Similar to these observation, Ali et al. (2019) reported
that the enrichment of pomegranate peel along with starch-based film
improved the antimicrobial activity against Staphylococcus aureus and
Salmonella. Even though, the incorporation of MPP (15%) gave a zone of
inhibition of 22.33 ± 2.51 mm against M. smegmatis, MP (15%) gave a
wider halo zone of 26.33 ± 1.70 mm showing a better activity. Against
the Gram-negative bacterium S. typhi, a zone of inhibition of 26.33 ±
1.70 mm was observed when MP film was incorporated. PP showed a
very less antibacterial activity compared to MP and MPP against
P. aeruginosa and no activity was observed against M. smegmatis and
S. typhi. Mango peels contains syringic acid, quercetin, mangiferin
pentoside and ellagic acid which are rich source of phenolic compounds
that exhibit antibacterial activity (Rehab et al., 2018; Vega-Vega et al.,
2013, Ajila, Aalami, Leelavathi, & Rao, 2010). The presences of phenolic
compounds act as antimicrobial agents and thus cause a change in
membrane permeability, with the release of ion gradients and pH ho­
meostasis. As a result, the cytoplasmic content in bacteria gets coagu­
lated accompanied by loss of normal cell metabolism and leads to
bacterial cell death (Raybaudi-Massilia, Mosqueda-Melgar, Sol­
iva-Fortuny, & Martín-Belloso, 2009).
3.12. Biodegradability
The biodegradability test evaluates the perishable potential of the
films in the environment by microorganism. A pictorial representation
of the experimental set up is depicted in Fig. 5 (a, b, c). All the films
including control were buried in the aluminium tray containing soil and
the degree of degradation of the films were estimated are shown in
Fig. 5d. Initial weights of the films were taken prior to the burial and
weight after degradation, after a span of 15 days, were noted (Sekhar
et al., 2016). It was observed that almost 50% of the films were degraded
after 15 days. The higher degradation of 80–90% was observed for lower
concentration (5% and 10%) of MP and MPP along with the loss of
shape, integrity and weight. The degradation rate of CSG with the
incorporation of (5% and 10%) MP and MPP increased up to 90%. This
Table 4
Antibacterial activity of CSG films incorporated with various concentration of
mango (puree and peel) and pineapple pomace.
Zone of inhibition (mm).
Samples
P. aueroginosa S. typhi M. smegmatis
CSG - - -
d -
5% MP 12.33 ± 0.47
–
10% MP 19.01 ± 0.81c -
–
15% MP 26.33 ± 1.25b 20.33 ± 1.24a 26.33 ± 1.70a
5% MPP 20.66 ± 0.58c
10% MPP 25.33 ± 1.52b
15% MPP 33.66 ± 0.58a 22.33 ± 2.51a 18.34 ± 1.52b
5% PP – – –
10% PP – – –
15% PP 14.33 ± 1.52d – –
a-d Means with different letters within a column are significantly different
(ANOVA/Turkey’s test, p < 0.05).
MP: Mango puree, MPP: Mango puree with peel and PP: Pineapple pomace.
‘-’ represents no zone of inhibition.
7
Food Bioscience 41 (2021) 100977
Fig. 4. Antibacterial test of 15% Mango puree (MP), Mango puree with peel
(MPP) and Pineapple pomace (PP) against Pseudomanas aeruginosaSample
1, 2, 3 and 4 represents CSG, MP, MPP and PP respectively.
was similar to the results reported by Nouraddini et al. (2018) where the
incorporation of eggplant flour (EF) improved the biodegradation of
starch film. The biodegradation time was comparatively slower for high
concentration of PP, MP and MPP, indicating that the increased amount
of bioactive compounds with antioxidant activity, such as phenolic
compounds, carotenoids and vitamins in mangoes and pineapples may
interfere with the microbial digestion of films, thereby inhibiting the
growth of microorganisms in the soil (Piñeros-Hernandez,
Medina-Jaramillo, López-Córdoba, & Goyanes, 2017). This was similar
to results reported by Medina Jaramillo, González Seligra, Goyanes,
Bernal, and Famá (2015) that the degradation time of cassava starch film
decreased with the addition of yerba mate extract.
4. Conclusion
An attempt was made to evaluate the potential of mango (pulp and
peel) and pineapple pomace for making biodegradable edible films. The
enrichment of CSG with these food by-products improved the thickness,
moisture content, swelling index and optic properties of the films. FTIR
analysis conferred that the compatibility between film constituents was
quite satisfactory and TGA analysis exhibited a similar decomposition
temperature for all the films, close to the degradation temperature of
native CSG films. The incorporation of high concentration of these fruit
components along with prebiotics like sorbitol (plasticizer) enhanced
the antioxidant and antimicrobial properties of the films, with pro­
nounced results observed for MPP films. By making such edible films, an
innovative process was developed for the utilization mango peel and
pineapple pomace which otherwise discarded or underutilized. Further
research has to be carried out to explore further formulations of these
materials and also to do in a larger scale to explore the possibilities to be
used in the food packaging industries. Application of this study in social
sector could generate additional income for the farmers and small and
medium enterprises in tropical areas.
Credit authorship contribution statement
Aliyath Susmitha: Conceptualization, Methodology, Formal anal­
ysis, Investigation, Writing - original draft, Keerthi Sasikumar: Meth­
odology, Writing - review & editing, Devi Rajan: Methodology,
Investigation, Arun PM: Methodology, Investigation, K Madhavan
Nampoothiri: Writing – Critical reading & editing, Supervision and
Project management. All authors have read and agreed to the published
version of the manuscript.
Author statement
We answered all the queries and incorporated the suggestions made
A. Susmitha et al.
Fig. 5. Biodegradability of CSG films incorporated with different concentration of MP, MPP and PP after 15 days of compost
a) Films prior to degradation
b) Weighed films placed on iron gauze
c) Films were buried in an aluminium tray containing soil. d) Films after 15 days degradation and percentage weight lost.
by the reviewers. A detailed response sheet also attached. All changes
were highlighted in yellow for easy evaluation of the changes brought in
to the revised version.
All authors agreed to the re submission of the revised manuscript to
the Journal and there is no conflict of Interest. There is no ethical issues
related to this manuscript Provided all the author’s contribution details
and also acknowledgd the funding.
Declaration of competing interest
All authors agreed to the submission of the manuscript to the Journal
and there is no conflict of interest.
Acknowledgement
Research fellowships from CSIR New Delhi are greatly appreciated
and the wok is carried out as a part of CSIR 800 program, socially useful
productive work. The authors sincerely acknowledge the technical help
provided by Mr. Brahma Kumar M and Mr. Peer Mohammed A in the
MSTD of CSIR NIIST for the material characterization studies.
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