Biocatalysis and Agricultural Biotechnology 35 (2021) 102096

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Biocatalysis and Agricultural Biotechnology

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

A novel sodium alginate active films functionalized with purple onion peel
extract (Allium cepa)
Luan Gustavo Santos, Gisele Fernanda Alves Silva, Bruno Marques Gomes,
Vilásia Guimarães Martins *
Laboratory of Food Technology, School of Chemistry and Food, Federal University of Rio Grande, Rio Grande, RS, 96203-900, Brazil

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

Keywords: The purple onion peel extracts (POPE) have a high content of phenolic compounds and can be used as a
Biodegradable polymer promising strategy in the development of active packaging. Biodegradable sodium alginate films have good
Active packaging characteristics of gel formation, flexibility, transparency, brightness, and low oil permeability. The incorporation
Antioxidant activity
of bioactive compounds from plant residues can be a low-cost technique to functionalize alginate-based mate­
Food waste
rials. The aim of this work was to evaluate the effects of POPE incorporation on the physical, mechanical,
barriers, antioxidants, and antimicrobial properties of alginate-based films. Filmogenic solutions were prepared
with a concentration of 0, 10, 20, and 30% of POPE and glycerol was used as plasticizer. The addition of POPE in
the sodium alginate matrix promoted the development of opaque red films, with a content of phenolic com­
pounds 43 times greater than the film without extract. As expected, the incorporation of POPE increased the
antioxidant activity and the thickness of the alginate-based films. In addition, the incorporation of phenolic
compounds promoted better interaction between the polymeric networks, reducing the water solubility of the
alginate films. The results indicated that the sodium alginate films incorporated with several concentration of
POPE have been shown to be an interesting material for the production of active packaging, being possible to
apply them in foods with high water activity or susceptible to lipid oxidation, to preserve and extend the shelf life
of these products.

1. Introduction (Paixão et al., 2019), with the ability to be decomposed by microor­

Tons of plastic are launched into nature year after year, aggravating
the environmental impacts due to the decomposition time of these ma­
terials (Thakur et al., 2018). However, due to their low cost and me­
chanical performance, synthetic polymers such as polyethylene,
polyvinyl chloride, polyethylene, polypropylene, polystyrene, or poly­
amide remain the materials most used as raw materials for food pack­
aging (Hanani, 2018; Kozłowicz et al., 2019). In the environmental
context, many types of research are being focused on replacing tradi­
tional fossil-derived polymers by renewable and biodegradable mate­
rials (Cazón et al., 2017; Nešić et al., 2020; Paixão et al., 2019).
Biodegradable films are defined as flexible films produced from one
or more biological materials, such as proteins, lipids, or polysaccharides
ganisms in a short time after use (Kozłowicz et al., 2019; Nešić et al.,
2020). Polysaccharides are biopolymers abundant in nature, presenting
low cost and good film-forming properties (Cazón et al., 2017; Hanani,
2018; Nešić et al., 2020).
Alginate is a polysaccharide extracted from microorganisms such as
Azotobacter and Pseudomonas or cell walls of brown algae (Phaeophy­
ceae), such as Ascophyllum odosum, Macrocystis pyrifera, and Ecklonia
cava (Senturk Parreidt et al., 2018). This polymer has great commercial
interest due to its potential to manufacture films and coatings with good
flexibility, brightness, water-solubility, emulsification capacity, and low
permeability to oil and oxygen (Oliveira Filho et al., 2019; Paixão et al.,
2019).
Technological advances related to the development of biomaterials

Abbreviations: POPE, Purple onion peel extract; CF, Control films; 10 EF, Film with 10% of extract; 20 EF, Film with 20% of extract; 30 EF, Film with 30% of
extract; RH, Relative humidity; TS, Tensile strength; EB, Elongation at break; WVP, Water vapor permeability; SW, Solubility in water; EF, Aqueous filmogenic
extracts; TPC, Total phenolics content; AT, Total anthocyanins content.
* Corresponding author.
E-mail addresses: luansantos.ea@outlook.com (L.G. Santos), giferalves@gmail.com (G.F.A. Silva), gmbruno95@gmail.com (B.M. Gomes), vilasiamartins@gmail.
com (V.G. Martins).

https://doi.org/10.1016/j.bcab.2021.102096
Received 1 April 2021; Received in revised form 6 July 2021; Accepted 7 July 2021
Available online 9 July 2021
1878-8181/© 2021 Elsevier Ltd. All rights reserved.
L.G. Santos et al.
applied as food packaging have been growing steadily. Several types of
research demonstrated that the incorporation of bioactive materials,
such as phenolic compounds, were able to improve mechanical and
barrier properties of films (Fabra et al., 2018; Hanani et al., 2019; Ju and
Song, 2020). The incorporation of natural antioxidants from plant ex­
tracts and essential oils in the polymeric matrix promotes the develop­
ment of active packaging, it can control the spoilage process and
increase the shelf life of food products (Romani et al., 2018).
Additionally, biocompounds have demonstrated the great potential
of antioxidant and antimicrobial activity, being a promising alternative
in the replacement of synthetic additives in active packaging. Several
studies show that when applied these active compounds in the devel­
opment of active packaging, these biocompounds can extend the shelf
life of fresh foods, minimally processed foods, and oils, delaying the
effects of spoilage (Guo et al., 2020; Paixão et al., 2019; Romani et al.,
2018).
Phenolic compounds are present mainly in essential oils, plant ex­
tracts, and food waste (Romani et al., 2018). The extraction of phenolic
compounds from plant matrix can be obtained through alcoholic,
organic, hydroalcoholic and aqueous solvents, in which, the polarity,
degree of polymerization and affinity with the matrix constituents, in­
crease the extraction efficiency of the biocompounds (Viera et al., 2017).
In this context, the purple onion peel (Allium cepa) is a waste that shows
great antioxidant and antimicrobial activity (Viera et al., 2017). These
activities are obtained from the biocompounds present in purple onion
peel extracts (POPE), mainly quercetins (Ju and Song, 2020) and an­
thocyanins (Lee et al., 2014), which can act in the capture of free radi­
cals and chelation of metals (Singh et al., 2009).
Previous studies show great efficiency in extracting bioactive com­
pounds from onion peel in acidified aqueous medium, obtaining extracts
with antimicrobial and antioxidant effects (Ali et al., 2016; Kabrah et al.,
2016; Singh, 2017). According to Makris and Kefalas (2015) acidified
aqueous media assist in the penetration of solvent into the cell walls of
plant matrices, enabling greater release of bioactive constituents,
obtaining low-cost, eco-friendly, and nontoxic phenolics-rich extract.
However, as far as we know, there are not studies related in the litera­
ture with the application of POPE in alginate-based films as an bioactive
agent.
Therefore, the present study aims to evaluate the effects on the
mechanical, physical, optical and barrier properties of sodium alginate-
based films functionalized with aqueous extracts of purple onion peel.
The results of this study may promote a new alternative for the devel­
opment of active biodegradable packaging capable of extending the
shelf life of foods by incorporating bioactive compounds derived from
food processing residues, such as purple onion peel.
2. Material and methods
2.1. Materials
Purple onion peels (Allium cepa var. Mercúrio) were obtained from
the local commerce of Rio Grande, Brazil. Sodium alginate P.A was
purchased from Dinamica (Indaiatuba, Brazil); The reagents ABTS (2, 2′ -
Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)); DPPH (2,2-
Diphenyl-1-picrylhydrazyl); Trolox (6-hydroxy-2, 5, 7, 8-tetramethyl­
chroman-2-carboxylic acid) and TPTZ (2,4,6-Tris(2-pyridyl)-s-triazine)
were obtained from Sigma-Aldrich (Sao Paulo, Brazil). All chemicals
reagent used were of analytical grade purity.
2.2. Extraction of phenolic compounds from purple onion peel
Purple onion peels were washed, dried in an oven with forced air
circulation (Fanem 520-C, São Paulo, Brazil) at 40 ◦ C for 4 h, then milled
into powders using a blender (Arno 700W-PL, São Paulo, Brazil) until
mesh 52 (0.355 mm). After, 10 g of powder was homogenized with 100
mL of distilled water acidified with acetic acid (pH 2.0) on an orbital
2
Biocatalysis and Agricultural Biotechnology 35 (2021) 102096
shaker (Tecnal TE-420, Piracicaba, Brazil) at 25 ◦ C for 1 h with stirring
of 150 rpm. In sequence, the content was centrifuged at 8709×g for 10
min and the supernatant was filtered in a paper filter under vacuum at
room temperature (25 ± 1 ◦ C), obtaining the POPE. The extract was
stored under refrigeration (5 ± 1 ◦ C) in amber glass and used on the
same day.
2.3. Production of active films
The sodium alginate films were prepared using the casting technique
(Oliveira Filho et al., 2019). The filmogenic solution was prepared ho­
mogenizing 2 g of sodium alginate in 100 mL of distilled water under
stirring of 1000 rpm for 1 h at 70 ◦ C. Then, 40% (w/w) of glycerol, based
on the dry weight of the polymer, was added and stirred for 20 min. The
solution was cooling to 40 ◦ C and added 10, 20, or 30% (v/v) of POPE,
homogenizing for 20 min. Petri dishes (Ø = 9 cm) were filled with 15 g
of filmogenic solution and dried in an oven with forced air circulation
(Quimis Q316M, Diadema, Brazil) at 40 ◦ C for 24 h. After drying, the
films remained protected from light 24 h in desiccators with relative
humidity (RH) of 55%. A saturated sodium chloride solution was used to
control the RH. The films developed with concentrations of 10, 20 and
30% (v/v) of POPE were denominated as 10 EF, 20 EF, and 30 EF,
respectively. A control film (CF) was obtained using the same process
but without adding POPE.
2.4. Characterization of the active films
2.4.1. Mechanical properties
The thickness is a fundamental property in the analysis of films,
being used in the calculation of the tensile strength and water vapor
permeability. The thickness of films was measured with a micrometer
(Insize IP54, precision 0.001 mm) at ten random positions for each film
sample.
Mechanical properties were evaluated using a texture analyzer
(Stable Micro Systems TA.XTplus, England). Tensile strength (TS) and
elongation at break (EB) of films were determined by ASTM Standard
Method D-882-02 (ASTM, 1995), where the films were cut in rectan­
gular strips 50 × 25 mm (length x width) and the test was made with
initial crosshead speed set at 1 mm/s and separation of 50 mm. The TS
and EB were calculated according to Eq. (1) and Eq. (2), respectively.
TS (MPa) =
Fmax
(1)
S
ΔL
EB (%) = x 100 (2)
L0
Where Fmax, S, ΔL, and L0 is the maximum force (N), initial cross-
sectional area of the film (m2), film elongation at break (mm) and the
initial gauge length of the specimen (mm), respectively.
2.4.2. Barrier properties
The water vapor permeability (WVP) was determined by the gravi­
metric method according to the ASTM Standard Method E96-00 (ASTM,
1995). In brief, plastic jars were filled with 15 g of anhydrous calcium
chloride (0% RH) and sealed with the active films. The jars were placed
in desiccators with 83% RH, using a saturated sodium chloride solution
to control the RH, and weighing them for 7 days at 25 ◦ C. The WVP was
measured by Eq. (3), where W is the weight gain (g); L is the thickness
(mm); A is the exposed area of the film (m2); t is the time of weight gain
(h); P is the vapor pressure difference between two sides of the film
(kPa).
( ) WxL
WVP g mm kPa− 1 day− 1 m− 2 = (3)
A x t x ΔP
The color and opacity were measured using a colorimeter (Minolta
CR 400, Tokyo, Japan) and used the CIELab color space, analyzing the
L.G. Santos et al.
parameter L (lightness/brightness), a* (redness/greenness), and b*
(yellowness/blueness). The colorimeter was calibrated with Illuminant
D65 (L* = 97.39, a* = 0,14 and b* = 1.94) and the total difference of
color (ΔE) was calculated according to Eq. (4).
ΔE = [(L*-L0)2 + (a*-a0)2 + (b*-b0)2]1/2 (4)
The opacity (%) of each film was determined by the ratio of the black
(Yb) and white (Yw) standards using the same equipment previously
mentioned.
2.4.3. Solubility in water (SW)
The SW of films was obtained according to Romani et al. (2018). The
film was cut in 2 cm of diameter and dry in an oven (DeLeo A15E, Porto
Alegre, Brazil) for 24 h at 105 ◦ C, then determining the initial dry weight
of the film (W0). The film was submerged in 50 mL of distilled water and
homogenized in an orbital shaker (Tecnal TE-420, Piracicaba, Brazil) at
150 rpm for 24 h. The films were dry again at 105 ◦ C for 24 h to
determine the final dry weight of the films (Wf). The solubility of the film
(%) was defined as the ratio between W0 and Wf.
2.5. Phenolic content
The total phenolic content and antioxidant activity of crude POPE
and functionalized sodium alginate films were evaluated. Water is the
main solvent used to simulate the migration and evaluate the antioxi­
dant activity of active compounds present in food packaging (Filipini
et al., 2020). Therefore, to evaluate the antioxidant of the films incor­
porated with POPE, aqueous film extracts (EF) were obtained, where,
0.1 g of the film was homogenized with 10 mL of distilled water in an
orbital shaker for 24 h at 25 ◦ C. Subsequently, the solution was centri­
fuged at 8709×g for 10 min and the solid particles were removed from
the supernatant using a vacuum filtration system at room temperature
(25 ± 1 ◦ C), obtaining EF. The same EF was used to determine total
phenolic content, ABTS, DPPH and FRAP.
2.5.1. Determination of total phenolic content
Total phenolics content (TPC) was obtained according to the meth­
odology described by Oliveira Filho et al. (2019). A test tube was filled
with 0.5 mL of POPE or EF, 2.5 mL of Folin-Ciocalteu 10% (v/v) and 2.0
mL of sodium carbonate solution 7.5% (w/v). The mixture was ho­
mogenized and incubated for 15 min at 50 ◦ C, then cooling until room
temperature (25 ± 1 ◦ C) and the absorbance was read in spectropho­
tometer UV-VIS (Kasuaki IL592, Brazil) at 760 nm. Standard curve of
gallic acid (0–65 μg/mL) was used to quantify the TPC and the content
was expressed as milligrams of gallic acid equivalents per gram of dry
sample (mg GAE/g).
2.5.2. Determination of total anthocyanin content
The anthocyanin content of the crude POPE was determined by the
pH-differential method (Lee et al., 2005). Briefly, a 500 μl aliquot of the
POPE was separately homogenized with 9.5 mL of potassium chloride
buffer (pH 1.0) or sodium acetate (pH 4.5). After 30 min of reaction, the
absorbance of the solutions was determined in a UV-VIS spectropho­
tometer (Kasuaki IL592, Brazil) at 510 and 700 nm. Anthocyanin total
(AT) was determined as mg of cyanidin-3-glucoside equivalent per 100 g
of purple onion peel (mg C3G/100 g) according to Eq. (5), where A510 nm
and A700 nm are the absorbance values at 510 and 700 nm, respectively,
FD is the dilution factor, MW is the molecular weight of the C3G (449.2
g/mol), V is the volume of the crude POPE (L), ϵ is the C3G molar
extinction coefficient (26,900 L/mol cm), L is the bucket length (1 cm)
and m is the mass of purple onion peel (g).
[ ]
(A510 nm − A700 nm )pH 1.0 − (A510 nm − A700 nm )pH 4.5 x FD x MW x 105 x V
AT =
εxLxm
(5)
3
Biocatalysis and Agricultural Biotechnology 35 (2021) 102096
2.5.3. Antioxidant activity assays
The antioxidant activity was determined by ABTS, DPPH, and FRAP
methods. The ABTS and DPPH assay were determinate using the method
described by Filipini et al. (2020). Briefly, ABTS (7 mM) and sodium
persulfate (140 mM) solution was mixed and kept 16 h at room tem­
perature (25 ± 1 ◦ C). Then, ABTS⋅+ radical solution was diluted with
ethyl alcohol until reach the absorbance of 0.70 ± 0.02 using a micro­
plate reader (Polaris Celer Biotecnologia S/A, Brazil) at 734 nm. An
aliquot (20 μL) of POPE or EF was homogenized with 280 μL of ABTS⋅+
radical in a microwell plate and incubated at 35 ◦ C for 20 min, obtaining
the absorbance of the extract (AE) at 734 nm.
The DPPH assay was prepared by mixing 20 μL of POPE or EF with
280 μL of ethanolic solution of DPPH radical (80 μM). The mixture was
incubated for 30 min at 35 ◦ C and the absorbance of the extract (AE) was
obtained at 517 nm in a microplate reader.
In all assays the same process was made replacing the extract with
distilled water, obtaining the control absorbance (Acontrol) of assays. The
ABTS and DPPH radical scavenging were calculated using Eq. (6).
AControl − AE
Scavenging effect (%) = (6)
Acontrol
The reduction of ion ferric (Fe3+) was evaluated using the FRAP
method according to Oliveira Filho et al. (2019). The FRAP reagent was
obtained by mixing 25 mL of acetate buffer (pH 3.6), 2.5 mL of TPTZ (10
mM) in HCl (40 mM) and 2.5 mL of FeCl3.6H2O (20 mM). In tubes were
homogenized 200 μL of the extract, 150 μL of distilled water, and 2.65
mL of reagent FRAP, incubating the mixture for 30 min at 37 ◦ C. The
absorbance values were read at 595 nm in spectrophotometer UV-VIS
and the results of the FRAP assay were expressed as Trolox equivalent
antioxidant capacity per gram of dry sample (mg TEAC/g) using a
standard curve of Trolox (0–80 μg/mL).
2.6. Antimicrobial properties of films
The agar diffusion method was performed to obtain the antimicrobial
activity of films against Escherichia coli O157:H7 and Staphylococcus
aureus ATCC 10832. Films were cut into a disc form of 15 mm diameter
using a circular knife and placed on brain heart infusion agar plates,
which had been inoculated with 100 μL (1.5 × 108 CFU/mL) of the cited
microorganisms. The plates were incubated at 37 ◦ C for 24 h and the
antimicrobial activities were obtained measuring the diameter (mm) of
the clear zone around the films using a pachymeter, all the experiments
were performed in triplicate.
2.7. Statistical analysis
The data were determined in triplicate and standard deviations were
reported. ANOVA was used to determine the analysis of variance and
Tukey’s test was applied to determine the differences among the prop­
erties of films in the 95% confidence.
3. Results and discussion
3.1. Barrier and mechanical properties of the films
The barrier and mechanical properties of food packaging play an
important role in preserving the quality of food products (Romani et al.,
2018). The properties as water vapor permeability, barrier gases, me­
chanical, optical, thermal, and microstructural properties vary accord­
ing to the composition of the films, related to the interaction of the
compounds present (Filipini et al., 2020). In Table 1, the mechanical and
barrier properties of the alginate-based films are presented.
Thickness evaluation is necessary because it is related to the per­
formance of the mechanical and barrier properties of the films (Borges
et al., 2015). In this study, it was observed that the films incorporated
L.G. Santos et al. Biocatalysis and Agricultural Biotechnology 35 (2021) 102096

Table 1 regions of the material (Filipini et al., 2020).

Thickness, Tensile strength (TS), Elongation at break (EB), Water vapor The CF showed an elongation of 4.91%, followed by a decrease of
permeability (WVP) and solubility in water (SW) of control film (CF) and films 25.25% with the addition of 30% of the extracts. The reductions in the
incorporated with 10, 20 and 30% (v/v) of POPE, called of 10 EF, 20 EF and 30 mechanical properties of the polysaccharide films incorporated into the
EF, respectively. phenolic extracts are reported in several studies in the literature (Biao
Films Thickness TS (MPa) EB (%) WVP (g.mm/ SW (%) et al., 2019; Liang et al., 2018; Liu et al., 2019; Moreno et al., 2020).
(mm) m2.day.kPa) Phenolic compounds can modify the thickness and microstructure of the
CF 0.049 ± 11.08 ± 4.91 ± 4.14 ± 0.02c 19.08 ± film, increasing the empty volume of the material, causing disorder and
0.002c 0.30a 0.20a 1.01a lesser interactions of the biopolymer chains (Liang et al., 2018; Liu et al.,
10 EF 0.055 ± 9.88 ± 4.31 ± 4.48 ± 0.10b 13.28 ±
2019; Ju and Song, 2020).
0.003b 0.67a 0.41ab 1.56b
20 EF 0.058 ± 7.83 ± 3.87 ± 4.61 ± 0.15b 13.99 ±
0.004ab 0.65b 0.16b 1.55b 3.2. Optical characteristics of the films
30 EF 0.062 ± 7.01 ± 3.67 ± 5.15 ± 0.19a 9.60 ±
0.004a 0.38b 0.30b 0.17c
The colors of the packaging are an important factor in the general
Mean ± standard deviation. Different letter in the same column indicates sig­ appearance of these materials and consumer acceptance (Nogueira et al.,
nificant differences (p < 0.05) by Tukey’s test. 2019). Results of the color and opacity parameters obtained for the so­
dium alginate films incorporated with different concentrations of POPE
with POPE presented greater thickness values when compared to the are shown in Table 2.
control film. This behavior is similar to several reported studies, in The incorporation of 10–30% of POPE on the biopolymer matrix
which, the presence of soluble solids from plant extracts, increases the promoted a significant difference (p < 0.05) of all color parameters
thickness of the material. Filipini et al. (2020) observed a 37.89% in­ about control film (CF). The decrease in L* values indicated that the
crease in the thickness of methylcellulose-based films by incorporating brightness of the films was decreased with the incorporation of POPE.
of jambolão skins extracts. Luo et al. (2019) incorporated guava leaf However, due to the presence of anthocyanins in POPE, the incorpora­
ethanolic extract in sodium alginate films and observed an increase of tion of the extract in sodium alginate films promoted an increase in
23.64% in the thickness of the films due the presence of soluble solids in parameter a* and reduction in parameter b*, obtaining films with red
the extract. color (Fig. 1).
As expected, the control film showed lower WVP. According to Oli­ Values of color difference (ΔE) and opacity increased with the
veira Filho et al. (2019), the incorporation of bioactive compounds in a addition of POPE. The films incorporated with the extract proved to be
sodium alginate matrix reduces the number of cross-linking sites, darker and more opaque than the CF. Similar behavior of color and
increasing the free volume, causing an increase in water vapor perme­ opacity parameters was reported by Nogueira et al. (2019) on arrowroot
ability. Similar results were observed by Shin et al. (2011) and Biao et al. starch films incorporated with anthocyanin-rich blackberry extracts,
(2019) incorporating extracts of grape seeds and green tea, respectively, showing a color difference of 62.33 after the addition of 40% (w/w) of
in alginate films. The authors attributed the increase in WVP to the loss extract in polymeric matrix.
of intermolecular interactions of the polymer and the increase in pore Darker packages with higher percentages of opacity are usually
size when extracts were added to the films. applied to fatty foods to extend the shelf life of these products, reducing
On the other hand, the high solubility of biodegradable films is one of the direct emission of light on these foods, avoiding the catalysis of lipid
the main disadvantages, hindering its use by industries. It was observed oxidation (Oliveira Filho et al., 2019).
that the solubility of the alginate films reduced by about 50.31% with
the incorporation of POPE (Table 1). The reduction in solubility may
3.3. Phenolics compounds content, antioxidant and antimicrobial activity
occur due to the presence of carboxylic acids in POPE. When combined
with alginate, carboxylic acids strengthen the bonds of biopolymeric
The POPE showed levels of phenolic compounds of 31.36 ± 0.37 mg
networks, reducing the solubility index (Kaewprachu et al., 2018).
GAE/g for TPC and 246.70 ± 4.24 mg C3G/100 g for AT. Similar values
Similar behavior was observed by Ju and Song (2020) when extracts of
of TPC and AT were obtained in extracts from different onion cultivars
yellow onion peel were added to funoran-based films, where the poly­
(Ali et al., 2016; Ju and Song, 2020; Viera et al., 2017), being these
phenols present showed greater interaction with the polymeric chains,
bioactive compounds the main responsible for the high antioxidant ac­
reducing the number of available hydroxyl groups that interact with the
tivity of the extracts in the FRAP assay (71.64 ± 0.60 mg TEAC/g).
water molecules, decreasing the solubility of the films.
Table 3 shows the results of the TPC and FRAP assay of the sodium
Besides, alginate films can be crosslinked from the reaction with
alginate films. The TPC content increased significantly (p < 0.05) with
monovalent cations, which are present in the chemical structure of an­
the incorporation of POPE in the sodium alginate films. Regarding the
thocyanins. The crosslinking process in alginate films increases the
films, 30 EF had the highest TPC content, being 43.32 times higher than
interaction between the biopolymer chains, reducing the solubility of
the material in water (Lopes et al., 2020). Films with less solubility are
Table 2
indicated mainly for fresh, frozen, or chilled food (e.g., fruit, meat
Color parameters of sodium alginate films incorporate with POPE, where CF is
products, dairy, and beverages), which have a high moisture content and
the control film, 10 EF, 20 EF and 30 EF are sodium alginate films incorporated
require insoluble packaging to maintain product integrity and quality with 10, 20 and 30% (v/v) of POPE, respectively.
(Nogueira and Martins, 2019).
Films L* a* b* Opacity (%)
Tensile strength and elongation at break are mechanical properties ΔE

that demonstrate the ability of the packaging to protect the physical CF 93.75 ± − 0.54 ± 7.91 ± 7.17 ± 10.59 ±
integrity of the food products (Hanani et al., 2019). The alginate films 0.23a 0.02d 0.12d 0.22d 0.31d
10 EF 58.24 22.22 ± 26.57 ± 51.39 ± 19.66 ±
incorporated with POPE had a reduction of up to 36.73% of TS in
±
0.38b 0.20c 0.13a 0.39c 0.62c
relation to CF. According to Luo et al. (2019) higher concentrations of 20 EF 39.44 ± 33.72 ± 24.87 ± 70.99 ± 32.48 ±
extracts incorporated in the polymeric matrix may cause a decrease in 0.24c 0.16b 0.39b 0.32b 0.74b
TS due to poor dispersion in the filmogenic solution. Moreover, films 30 EF 36.49 ± 34.76 ± 22.41 ± 73.79 ± 45.85 ±
0.12d 0.21a 0.53c 0.44a 0.80a
with lower TS values indicate that the material has a greater free volume
and pores in the structure, caused by the reduction of the crystalline Mean ± standard deviation. Different letter in the same column indicates sig­
nificance differences (p < 0.05) by Tukey’s test.

4
L.G. Santos et al.
Fig. 1. Visual aspects of the control films (CF) and sodium alginate films
incorporated with 10, 20 and 30% (v/v) of POPE, called of 10 EF, 20 EF and 30
EF, respectively.
Table 3
Total phenolics content (TPC) and ferric reducing ability of plasma (FRAP) of the
sodium alginate films, being CF, 10 EF, 20 EF and 30 EF are sodium alginate
films incorporated with 0, 10, 20 and 30% (v/v) of POPE, respectively.
Sample TPC (mg GAE/g film) FRAP (mg TEAC/g film)
CF 0.59 ± 0.28d 3.19 ± 0.15d
10 EF 19.93 ± 0.32c 23.86 ± 1.00c
20 EF 20.86 ± 0.16b 52.30 ± 0.81b
30 EF 25.56 ± 0.33a 55.33 ± 1.07a
Mean ± standard deviation. Different letter in the same column indicates sig­
nificance differences (p < 0.05) by Tukey’s test.
the CF film. The 10 EF and 20 EF films, the TPC content was 22.02 and
18.39% lower than the 30 EF film, respectively.
In the evaluation of the antioxidant activity by the FRAP method, the
films incorporated with POPE obtained values ranging from 23.86 to
55.33 mg TEAC/g. Due to the higher TPC content, the highest antioxi­
dant activity was obtained in the 30 EF film, corresponding to about
77.23% of the antioxidant activity by the FRAP method of the crude
POPE. According to Lee et al. (2014) the main compounds of POPE are
quercetins and anthocyanins which are shown to be powerful antioxi­
dants due to their mechanism of reducing free radicals, chelating metal
ions, and inhibiting oxidation.
The ABTS and DPPH method is widely used to assess the antioxidant
activity of hydrophilic and lipophilic biocompounds, respectively (Oli­
veira Filho et al., 2019). Fig. 2 shows the evaluation of the scavenging of
the radicals ABTS and DPPH of the POPE and the films developed. The
30 EF film showed the highest percentage of reduction of radicals ABTS
and DPPH, being 98.17% and 48.20%, respectively, where, similar ABTS
values were obtained in 30 EF and crude POPE. The content of phenolic
compounds present in plant extract increase the antioxidant activity of
biopolymer films and it is directly related to the concentration of ex­
tracts incorporated in the biopolymer matrix (Fabra et al., 2018; Oli­
veira Filho et al., 2019).
On the other hand, the scavenging activity of DPPH on the POPE and
films showed lower values when compared with the antioxidant activity
by the ABTS method. This is because the bioactive compounds present in
the POPE, such as anthocyanins and quercetins, present hydrophilic
5
Biocatalysis and Agricultural Biotechnology 35 (2021) 102096
Fig. 2. ABTS and DPPH scavenging activity of the purple onion peel extract
(POPE) and sodium alginate films incorporated with 0, 10, 20 and 30% (v/v) of
POPE, abbreviated as CF, 10 EF, 20 EF and 30 EF, respectively. Uppercase and
lowercase letters indicate different analyzes. Different letters for the same
method indicate statistical difference (p < 0.05).
behavior and the DPPH radical demonstrates greater interaction with
lipophilic antioxidant compounds (Oliveira Filho et al., 2019). Bio­
compounds act mainly in the capture of free radicals, preventing
oxidation when incorporated in active packaging. This packaging sys­
tem becomes very promising due to the reduction in the use of synthetic
additives and the extension of the shelf life of food products (Romani
et al., 2018).
The films produced did not show antimicrobial activity for the mi­
croorganisms studied. Similar results were obtained by Viera et al.
(2017) and Kabrah et al. (2016) where aqueous extracts of purple onion
peel did not show an inhibition zone for the microorganisms S. aureus
and E. coli. This result can be influenced by the solvent used in the
extraction or by the low content of phenolic compounds present in the
dry films, being insufficient to inhibit the growth of the tested
microorganisms.
Despite this, sodium alginate films are non-toxic materials, with
good physical and structural characteristics that, by incorporating
phenolic extracts from a food residue (POPE), are functionalized,
reducing the percentage of solubility and increasing its antioxidant ac­
tivity. Thus, the material developed in this study can be applied as an
active packaging, reducing, and preventing the oxidation of packaged
foods.
4. Conclusion
The presence of phenolic compounds in POPE promoted the devel­
opment of active films based on sodium alginate with opaque red color
and higher antioxidant activity, indicating great potential for applica­
tion in foods susceptible to lipid oxidation (e.g., oil and fats). Moreover,
the 30 EF film showed less solubility in water, which can be justified by
the interactions of the anthocyanidin cations with the alginate poly­
meric network, providing the crosslinking of the material. The reduction
in the water solubility of alginate films allows the application of this
material as packaging in foods with high water activity or when the film
remains in contact with the food water during storage, avoiding the
exudation of processed or fresh products. Further experiments are
necessary to improve the mechanical properties of the films, also it is
important to carry out the application and evaluation of the shelf life of
fatty foods packaged with those films.
L.G. Santos et al.
CRediT authors
Luan Gustavo Santos: Conceptualization; Data curation; Formal
analysis, Investigation; Methodology; Roles/Writing - original draft.
Gisele Fernanda Alves Silva: Conceptualization; Data curation; Formal
analysis, Investigation; Methodology; Roles/Writing - original draft.
Bruno Marques Gomes: Formal analysis, Investigation. Vilásia
Guimarães Martins: Project administration; Supervision; Writing - re­
view & editing.
Declaration of competing interest
None.
Acknowledgments
This study was financed in part by the Coordenação de Aperfeiçoa­
mento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001
and Conselho Nacional de Desenvolvimento Científico e Tecnológico
(CNPq). This research was developed within the scope of the Capes-PrInt
Program (Process # 88887.310848/2018-00).
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