International Journal of Biological Macromolecules 226 (2023) 1352–1359

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

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

Active packaging of corn starch with pectin extract and essential oil of
Turmeric Longa Linn: Preparation, characterization and application in
sliced bread
Maria Nicheilly Pontes Araújo a, Cristiani Viegas Brandão Grisi b, Cybelle Rodrigues Duarte a, c,
Yeda Medeiros Bastos de Almeida a, Glória Maria Vinhas a, *
a
Federal University of Pernambuco, Postgraduate Program in Materials Science, 50740-560 Recife, PE, Brazil
b
Federal University of Paraiba, Postgraduate Program in Agro-Food Technology, 58220-000, Bananeiras, PB, Brazil
c
Federal University of São Carlos, Graduate Program in Materials Science and Engineering, 13565-905 São Carlos, SP, Brazil

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

Keywords: The use of active packaging to reduce food waste has been a very effective alternative. An eminent concern is the
Corn starch use of plastic materials of petroleum origin and toxic additives in the processing of these packages. Thus, the
Pectin extract focus on the use of biodegradable and natural raw materials that minimize waste generation and promote greater
Turmeric essential oil
consumer safety has been preferable. The objective of the research was to investigate the effects of turmeric
Active packaging
essential oil (TEO) on corn starch and pectin extract films manufactured by solution casting method. The anti­
oxidant and antimicrobial potential of the oil was confirmed by the tests: antimicrobial diffusion disk, deter­
mination of the content of phenolic compounds and antioxidant activity by the DPPH and FRAP method. The
chromatographic analysis confirmed the presence of active chemical constituents such as Turmerone, Ar-
Turmerone and β–Turmerone. The results showed that the oil promoted a change in the color of the films,
increased mechanical strength and reduced flexibility, keeping transparency, solubility, WVP and thermal sta­
bility unchanged. In the direct application test of the film as packaging for sliced bread, no visible contamination
was detected during the nine weeks of analysis. Therefore, the active film with 3 % TEO was shown to be a viable
solution for manufacturing biodegradable and safe active films that can be applied as food packaging.

1. Introduction genera Aspergillus, Chrysonilia, Cladosporium, Curvularia, Eurotium,

Packaging plays a fundamental role in the food industry. Their main
applications are for containment, protection, preservation, and
adequate transport, which essentially helps to reduce losses and global
waste in the food sector [1]. The direct connection between these in­
dustrial sectors makes it possible to understand the reasons that make
the food industry the main end user of the packaging sector, corre­
sponding to a consumption of 40 % of the plastic produced [2,3].
Waste in the food sector is especially considerable in bakery products
because bread reaches its expiration date shortly, in a period of days.
Thus, it cannot be completely consumed, most of the time being dis­
carded after it expires [4]. This problem was associated with the rela­
tively short shelf life of bread, at room temperature (25 ◦ C), due to high
water activity and acidic pH, which makes them susceptible to
contamination by various microorganisms. In general, fungi of the
Fusarium, Penicillium, Rhizopus e Mucor are the causes of bread spoilage.
The action of fungi on bread is diverse, the most noticeable aspects of
change are color, taste, and odor of the product [5].
The search for new materials such as active packaging, capable of
increasing the shelf life of food and reducing the quantity of pre­
servatives added in food formulations, has represented a major tech­
nological advance for the food industry [6]. The technology used in
these packages makes use of additives that contain chemical constitu­
ents in their formulation capable of developing antimicrobial and anti­
oxidant action, in addition to polymers and other essential additives,
such as plasticizers [7,8].
The selection of natural and biodegradable raw materials to produce
these packages has been preferable, as a result of the environmental
problems generated using conventional polymers of petrochemical and
non-biodegradable origin. Additionally, the use of agro-food waste as

* Correspondence to: G.M. Vinhasa, Federal University of Pernambuco, 50740-560, Recife, PE, Brazil.
E-mail address: gmvinhas@yahoo.com.br (G.M. Vinhas).

https://doi.org/10.1016/j.ijbiomac.2022.11.248
Received 22 July 2022; Received in revised form 18 November 2022; Accepted 23 November 2022
Available online 28 November 2022
0141-8130/© 2022 Published by Elsevier B.V.
M.N.P. Araújo et al.
raw material in the production of these packaging reduces production
costs [9].
Due to the biocompatibility, abundance, low cost, functional, phys­
ical and chemical properties that the starch constituents present, this has
been a suggested material in the application of active packaging.
However, their use needs to be well investigated, as they have me­
chanical, thermal and barrier properties lower than petroleum derived
polymers [10,11].
Natural additives such as essential oils have been used as an effective
alternate to replace the chemical preservatives to avoid the risks of
toxicity and insecurity of consumers in case of excessive consumption
[12]. Essential oils are substances classified as Generally Regarded as
Safe (GRAS), which guarantees its use in food products, its use as anti­
oxidant and antimicrobial additives in packaging material has been
applied due to its high content of terpenes and phenolic compounds
[13,14]. In particular, turmeric essential oil has been highlighted
because of its content of phenolic acids (turmerone, zingiberene, etc.),
flavonoids (myricetin, fisetin, morin, etc.) and curcuminoids (curcumin,
dimethoxy curcumin, bis-dimethoxy curcumin, etc.) that have biological
properties already recognized as antioxidant, anti-inflammatory, anti-
cancer, anti-arthritic, anti-depressant, anti-diabetic, and antimicrobial
[15].
In the literature, there are studies that used corn starch, pectin, and
glycerol as a polymer matrix in the development of active packaging
systems through the incorporation of synthetic additive, oil essential,
and plant extracts [12,16–19]. To the best of our knowledge, there are
no studies in the literature using the turmeric essential oil additive with
antimicrobial and antioxidant activity in the polymer matrix of corn
starch, glycerol, and passion fruit peel pectin extract film.
So, the objective of this research was to develop a film of corn starch
added with glycerol, passion fruit peel pectin extract, and turmeric
essential oil, capable of promoting antimicrobial and antioxidant ac­
tivity to be applied as an active film for food packaging. Firstly, the ef­
fects of turmeric essential oil were studied and its potential as an active
additive was analyzed, then the films were prepared by solution casting
method. The thermal, mechanical, barrier and optical properties of the
films were evaluated, as well as an application test as a bread wrapper to
evaluate its effectiveness as a fungicide.
2. Materials and methods
2.1. Materials
Corn starch (Maizena brand) was obtained from the Recife-PE mar­
ket. The glycerol was obtained from Química Moderna. The turmeric
essential oil (Terra flor brand), originating in India (IBD - Brazil natural
ingredients certification) was acquired in a compounding pharmacy.
Passion fruit was acquired in its initial stage of development in the local
market of Recife (8◦ 04′ 03′′ S and 34◦ 55′ 00′′ W), in the period Octo­
ber–November 2019.
The pectin extract (PE) was obtained from the firm and well washed
passion fruit. The total of 200 g of the passion fruit peel was weighed and
added in Becker containing 500 mL of distilled water, which was put on
heating until the material softened and became translucent. Subse­
quently, 20 mL of lemon was added and boiled for 10 min. After cooling,
the liquid was extracted under filtration and stored in a refrigerator
(5 ◦ C).
2.2. Film preparation
The films were made by solution casting technique. Initially, the
starch, pectin extract, glycerol and distilled water were mixed manually
for 1 min. Then, the mixture was stirred with the aid of a mixer at 1000
rpm, on a water bath at 80 ◦ C for 8 min, aiming for starch gelatinization.
After this, turmeric essential oil (TEO) was dissolved in 2 mL of ethanol
and added to the solution, in which it remained under stirring in a mixer
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International Journal of Biological Macromolecules 226 (2023) 1352–1359
at 1000 rpm for 2 min. After preparing the film-forming solution, it was
allowed chill until 45 ◦ C, then poured into acrylic Petri dishes (140 × 15
mm) and dried in an air circulation oven at 40 ◦ C for 7 h. Then they were
conditioned (75 % RH/25 ◦ C). Each prepared solution had a total mass
of 60 g, comprising the following compositions: 3 % m/m CS, 30 % of the
PE and glycerol in relation to the starch mass, 2 mL of ethanol, distilled
water and 1 %, 2 % and 3 % of the TEO in relation to the starch mass.
The coding given to the developed films were CS, CS – 1%TEO, CS – 2%
TEO, CS – 3%TEO. The composition for each film formulation is listed in
Table 1.
2.3. Characterization of turmeric essential oil
The total phenolic content was determined by the method of
Singleton et al. [20]. Initially, 10 mg of the sample was diluted in
ethanol, using a 10 mL volumetric flask. An aliquot of 150 μL of the
solution was transferred to a test tube, together with 2610 μL of distilled
water and 60 μL of Folin Ciocalteau reagent, which remained under
agitation for 60 s. After this time, 180 μL of Na2CO3 solution (15 %) was
added and the solution was stirred for another 30 s. After 120 min of
reaction at rest, in the absence of light, the reading was performed in a
spectrophotometer (Edutec) at 760 nm, in triplicate. A standard curve
with gallic acid at concentrations from 1 to 20 mg/mL was obtained
under the same conditions (y = 0.0924x + 0.1222, R2 = 0.986). The
total phenolic content was expressed in mg of GAE (gallic acid equiva­
lent) per gram of the sample.
The determination of antioxidant activity by the free radical method
2,2-diphenyl-1-picrylhydrazyl (DPPH) was performed to the methodol­
ogy developed by Brand-Williams et al. [21]. The mixture was carried
out by adding 150 μL of the TEO solution (1 mg. mL− 1), 150 μL of
ethanol and 2700 μL of the DPPH solution. The DPPH solution in ethanol
was used as a control. After 30 min of incubation of the mixture at room
temperature (25 ◦ C), the samples were read in a UV–visible spectro­
photometer at 517 nm, with the results expressed in percentage of in­
hibition (%I).
The antioxidant activity measured by the Ferric Reducing Antioxi­
dant method was performed to the methodology of Rufino et al. [22].
The FRAP reagent was prepared by mixing 25 mL of acetate buffer (0.3
M, pH 3.6), together with 2.5 mL of the TPTZ solution (2,4,6-tri (2-
pyridyl-1,3,5 triazine) (10 mM) and 2.5 mL of aqueous ferric chloride
solution (20 mM). Then, an aliquot of 90 μL of the sample was added to a
test tube, together with 270 μL of distilled water, and 2700 μL of the
FRAP reagent. The test tubes were placed in a water bath at 37 ◦ C for 30
min. A standard curve for ferrous sulfate (500 to 2000 μM) was con­
structed. Results were expressed as μmol ferrous sulfate per gram of oil.
The determination of antibacterial activity followed the methodol­
ogy described by Silva et al. [23], testing Staphylococcus Aureus,
Escherichia coli and Enterobacter bacteria, in triplicate. The culture me­
dium (Nutritive Agar) was prepared and poured into a sterile Petri dish.
After solidification of the medium, a 0.1 mL aliquot of the microbial
Table 1
Formulations of films.
Films Starch PE Glycerol Ethanol Water TEO
(g) (g) (g) (mL) (mL) (g)
CS 1.80 0.54 0.54 – 57.12
CS – 1 % 1.80 0.54 0.54 2.00 55.10 0.018
TEO
CS – 2 % 1.80 0.54 0.54 2.00 55.08 0.036
TEO
CS – 3 % 1.80 0.54 0.54 2.00 55.06 0.054
TEO
CS (corn starch/pectin extract [PE]). CS – 1 % TEO (corn starch/pectin extract/
1 % turmeric essential oil [TEO]). CS – 2 % TEO (corn starch/pectin extract/2 %
turmeric essential oil). CS – 3 % TEO (corn starch/pectin extract/3 % turmeric
essential oil).
M.N.P. Araújo et al.
suspension of the bacteria (standardized to the McFarland scale) was
spread over this medium. Then, TEO impregnated paper discs were
transferred to the surface of the culture medium with the inoculated
bacteria. The plate was placed in an oven at 35 ◦ C for 48 h and the in­
hibition halo was measured with the aid of a ruler.
2.4. Characterization of active films
2.4.1. Solubility and water vapor permeability (WVP) of films
The solubility was determined following the methodology described
by Nordin et al. [16], in triplicate. The films (2 cm × 2 cm) were dried in
an air circulation oven at 105 ◦ C for 24 h, and then weighed to obtain the
initial mass (Im). Then, they were immersed in 50 mL of distilled water
and kept at rest for 24 h. Subsequently, the samples were filtered and
subjected to drying in an oven at 105 ◦ C for 24 h again and then weighed
in order to obtain the final dry mass (Dm). The solubility was given by
Eq. (1):
S (%) = [(Im − Dm)/Im ] × 100 (1)
The WVP was established using gravimetric method E96–95 of the
American Society for Testing and Materials [24]. Square samples (6 cm
× 6 cm) were placed properly sealed, forming a permeation area (7.069
× 10− 4 m2). These containers were filled with calcium chloride (CaCl2)
up to a distance of approximately 1.5 cm of the film. Then, the con­
tainers were placed in a desiccator containing saturated sodium chloride
(NaCl) solution (75 % RH). Weight gain was monitored every 24 h for six
days. The WVP was measured according to Eq. (2):
WVP = (Δm/A.Δt) × (X/(Ps × (RH1 − RH2) ) (2)
where, WVP is water vapor permeability, Δm/Δt is the slope of the
weight gain curve per unit time, A is the exposed surface area of the film
(m2), X is the average film thickness, Ps is the saturation pressure of the
water, RH1 is the relative humidity inside the desiccator and RH2 is the
relative humidity inside the container.
2.4.2. Mechanical properties
Tensile strength (TS) and percentage elongation (ε) at break were
measured using a SHIMADZU static testing instrument-AGSLine-X10Kn,
operating according to ASTM D882-12 specifications [25]. The speci­
mens (80 × 15 mm) had their thickness measured with a stainless-steel
digital caliper (LEE TOOLS), in five random positions in all samples,
previously conditioned at 75 % RH and 25 ◦ C for 7 days. The specimens
were fixed with an initial grip separation distance of 50 mm and a grip
separation speed of 12.5 mm/min. The TS (MPa) was determined by the
relationship between the maximum tensile force measured (σ) and the
initial cross-sectional area of the specimen (Ao), according to Eq. (3).
The ε (%) was determined by the relationship between the distance
traveled in the displacement of the grips until the break [ΔL = final
length (Lf) − initial length (Lo)] and initial length of the specimen (Lo),
according to Eq. (4).
TS (MPa) = σ/Ao (3)
ε (%) = (ΔL/Lo) × 100 (4)
2.4.3. Thermogravimetric analysis
Thermogravimetric analysis was performed using the TGA-50 SHI­
MADZU equipment, with a temperature range from 25 to 550 ◦ C, with a
heating rate of 10 ◦ C/min under a nitrogen atmosphere, with a sample
mass that varied in around 8 to 10 mg and nitrogen flow of 50 mL/min.
2.4.4. Fourier transform infrared spectroscopy (FTIR) and principal
component analysis (PCA)
The films were analyzed using Infrared Spectroscopy from SHI­
MADZU, model IRTRACER100, using the attenuated reflectance method
(ATR). The spectra were obtained according to the methodology of
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International Journal of Biological Macromolecules 226 (2023) 1352–1359
Jaramillo et al. [26] in transmittance mode, in the range of 4000 to 400
cm− 1, with a resolution of 4 cm− 1 and 45 scans. PCA was performed
based on the spectra obtained in the infrared, in which each film sample
was analyzed at five different points, obtaining as a result of multivar­
iate data a matrix composed of 20 spectra. The Unscrambler 9.7 software
was used for data processing.
2.4.5. Transparency and colorimetry
The transparency was analyzed with a UV–Vis spectrophotometer in
the range of 600 nm and determined according to Eq. (5) [27]. Where, %
T, is the percentage of transmittance and b, is the film thickness (mm).
T = Log (%T)/b (5)
The colorimetric analysis was performed following the methodology
described by the ASTM E308-17 [28]. The experiment was carried out
with GretagMacbeth - Color-Eye 2180 colorimeter, the CIELAB Ttan D65
system, reading with an angle of 10◦ and a viewing area of one square
inch. The films were placed on a standard white plate (L* = 95.833; a*
= − 0.22; b* = 2.35). The total color difference (ΔE) calculated by Eq.
(6) with respect to the control formulation (without TEO).
[ ]1/2
ΔE* = (ΔL* )2 + (Δa* )2 + (Δb* )2 (6)
2.4.6. Application of the film on the loaf of bread
The films (10 cm × 6 cm) were used to package slices of bread in the
dimensions 2 cm × 2 cm. The packages were properly sealed and stored
at room temperature (25 ◦ C). The evaluation of the appearance of the
fungus in the breads was performed visually. The packaged bread was
monitored weekly (Fig. 2S) by observing any noticeable change in color
[29]. Cellophane was used as a control material. The loaf of bread tested
was the traditional brand “Pão e Mel Indústria de Panificação LTDA-
ME”, purchased at the local market in Recife - PE, valid for 10 days.
2.5. Statistical analysis
The statistical analysis of the results aimed to verify the degree of
dispersion and the reliability of the data. Data were analyzed by Analysis
of Variance (ANOVA) and Duncan’s test was applied at 5 % probability
using ASSISTAT software version 7.7 [30].
3. Results and discussion
3.1. Turmeric essential oil (TEO)
The TEO presented 49.20 mg GAE/g in terms of total phenolic
compounds and antioxidant activity, of 34.01 ± 1.76 % by DPPH free
radical inhibition, and 28.05 ± 0.06 mmol Fe2SO4/g, by the FRAP
method. The chemical compounds identified in the TEO (Table 1S)
belong to the sesquiterpenoid groups (Ar–Turmerone, β-Turmerone and
Turmerone) and sesquiterpenes (Ar-curcumene, α-Zingiberene and
β-sesquiphelandrene), whose predominant composition (73.51 %) was
Turmerone (C15H22O), Ar–Turmerone (C15H20O) and β-Turmerone
(C15H22O), which are potential compounds in the development of
antioxidant activity.
The results determined in this study are in agreement with those
observed in the literature by Aggarwal et al. [31] who determined that
among the major compounds in TEO are ar-turmerone (17.9 %) and
α-turmerone (14.6 %); Gounder and Lingamallu [32] found ar-
turmerone (21.0 to 30.3 %), α-turmerone (24.8 to 33.5 %), β-turmer­
one (18.9 to 21.1 %), ar -curcumene (1.2 to 1.9 %), α-Zingiberene (2.2 to
2.6 %) and β-Sesquiphelandrene (1.8 to 2.4 %); Li et al. [33] also found
Ar-Turmerone as the predominant compound in the oil.
The variation in the percentage of the various chemical compounds
identified in the TEO in relation to the various presented studies comes
from factors intrinsic to the plant, such as: geographic location of origin,
climatic conditions, stage of development or age of the plant, among
M.N.P. Araújo et al. International Journal of Biological Macromolecules 226 (2023) 1352–1359

others. Furthermore, the volatility of essential oil chemical components,
oxidation of chemical compounds, extraction methods and some other
factors associated with the oil are responsible for causing differences in
the composition and content of turmeric essential oil [32,34].
The TEO was able to inhibit the growth of the Staphylococcus Aureus
(12 mm of inhibition halo) and did not present an inhibition halo for
Enterobacter and Escherichia coli, which confirms the antimicrobial
power that the TEO has against gram positive bacteria. The results of this
study corroborate the literature that evidenced a more effective action of
TEO with gram positive bacteria. That it can be explained by the absence
of the outer membrane in these bacteria, which makes them unable to
limit the entrance of exogenous substances. Franco et al. [35] evaluating
the activity of turmeric essential oil verified the antibacterial action on
S.aureus, but it did not show any activity on the other microorganisms
tested, Salmonella choleraeseus and E. coli. These results were an indi­
cation that this oil has substances with antioxidant and antimicrobial
potential, which may be of interest for application in antioxidant active
packaging.
3.2. Solubility and water vapor permeability (WVP)
The solubility also remained unchanged with the addition of oil
(Table 2). The interaction of the oil with the polymer matrix reduces the
solubility of the films in water, due to the hydrophobicity of the oil
molecules. However, the result of this study showed a non-significant
reduction. The WVP of films ranged from (5.11 to 6.35) × 10− 7 g/
h⋅m⋅Pa, with no significant changes after the addition of the additive.
This behavior is acceptable, as it is not always possible to observe
changes in this property from the addition of essential oils. Di Giuseppe
[36] when adding rosemary essential oil to the chitosan matrix also did
not observe changes in the WVP of the films. Li et al. [19] observed an
increase in WVP when incorporating a higher concentration of TEO in a
chitosan matrix. The change in WVP depends on factors such as: char­
acteristics and concentration of the additive, and the oil/matrix
interaction.
3.3. Mechanical properties
The effect of TEO on the tensile strength and flexibility of the films
were significant (Table 2). The thickness of the films varied between
0.094 and 0.103 mm. The TS increased after the addition of the additive,
except for the film with 2 % TEO. The film with 3 % oil showed the
highest value for mechanical strength. Flexibility, in general, was
reduced with the addition of higher concentrations of TEO. Such a
reduction is associated with the hydrophobicity of the oil in a hydro­
philic matrix, as changes in mechanical properties caused by the addi­
tion of hydrophobic substances can promote the formation of a structure
with lower mobility [37]. Another reason be associated with the
reduction of the water content present in the film after the addition of
the TEO, which reduces the plasticizing action of the water in the film
[19].
The results of this study are in accordance with those presented by
Souza et al. [38] synthesized CS films containing Pickering emulsions
and wood essential oil and significantly improved the mechanical
strength of the film with strength values of 10.8 MPa and flexibility of
29.5 %. The films produced showed greater flexibility when compared
to synthetic polymer films, polypropylene (PP) and poly (ethylene
terephthalate) (PET) produced by Lee et al. [39]. In the results, the oil-
free films presented an elongation value of 50.67 %, while the film
containing star anise essential oil presented a value of 45 %.
It was also important to consider that during the production of the
films, analyzing macroscopically. It was only possible to obtain films
without bubbles and cracks after the addition of the PE, which made
possible to affirm that the pectic compounds acted promoting greater
stability to the films. The reason for this increased stability was
explained by the three-dimensional molecular structure of pectin, which
has, in its side chains, free polar groups such as hydroxyls. Pectin mol­
ecules promoted greater aggregation of particles in solution, favoring
the formation of gels, stabilization of solutions and several functional
properties [40]. Thus, the use of the plasticizer in association with the
PE also contributed to improve the mechanical properties of the films.

3.4. Thermogravimetric analysis

Table 2
Structural properties of films: Water Vapor Permeability (WVP); Solubility; The behavior of the TG and DTG curves (Fig. 1) was similar for all
Tensile Strength, Elongation at break (Ɛ), Transparency and Colorimetry of formulations of the prepared films, presenting three stages of mass loss
films. characteristic of starch films [41]. The first stage of mass loss (25 to
250 ◦ C) was caused by the reduction of free water in the films because of
Films CS CS – 1%TEO CS – 2%TEO CS – 3%TEO
water evaporation. In addition, at this stage, glycerol degradation and
WVP 6.35a ± 1.69 6.26a ± 1.13 5.46a ± 1.04 5.11a ± 4.87
evaporation of TEO began [42]. The second stage occurs between 280 ◦ C
(10− 7 g/
h⋅m⋅Pa)
and 350 ◦ C. In this temperature range, glycerol degradation continued
Solubility (%) 28.74 a ± 27.33a ± 27.44a ± 27.37a ± and the beginning of thermal decomposition of starch was observed
2.51 9.26 9.07 3.39 [38]. In the third step, after 350 ◦ C, a loss of mass attributed to starch
TS (MPa) 5.40c ± 0.79 8.48b ± 0.71 5.39c ± 0.54 10.94a ± decomposition was observed. Studies attribute to this stage, elimination
1.11
of hydrogen groups, decomposition, depolymerization of starch carbon
Ɛ (%) 66.97b ± 92.25a ± 54.25d ± 61.85c ±
2.89 3.62 1.32 0.60 chains and loss of mass generated by the disintegration of carbonaceous
Transparency 18.717a ± 13.590a ± 15.370a ± 16.993a ± residues [16,42].
(Log (%T)/ 2.917 2.640 3.300 1.484 Despite this similarity, the CS-3 % TEO formulation shows a subtle
mm)
improvement in stability from approximately a temperature of 170 ◦ C.
L* 94.66ª ± 94.62ª ± 94.15b ± 93.88c ±
0.27 0.09 0.17 0.15
Probably at this concentration there was a greater intermolecular
a* − 0.58d ± − 3.97c ± − 6.00b ± − 6.66ª ± interaction between the oil components and the polymeric matrix fa­
0.05 0.42 0.45 0.40 voring this event. A similar result was obtained by Cai et al. [43] who, in
b* 3.59d ± 0.17 12.53c ± 19.11b ± 22.08a ± their formulation with a higher percentage of thyme oil in the corn
1.25 1.44 1.47
starch matrix, observed a slight change in the thermogravimetric
ΔE* – 09.57c ± 16.45b ± 19.49a ±
1.39 1.51 1.40 analysis.
In a general analysis of this result, it is possible to state that the
addition of TEO did not change the thermal stability of the CS films, due
to the similarity observed in the TG mass loss curves. By analyzing the
maximum temperature of degradation, observed in the DTG, it was not
Values expressed in mean (n = 3) ± standard deviation. The means followed by possible to verify significant differences in the depth and width of the
the same letter on the same line do not differ statistically from each other by the peak, proving that the oil did not change the thermal stability of the
Duncan Test at the level of 5 % probability (p < 0.05). films. Di Giuseppe [36] when evaluating the effect of essential oils on

1355
M.N.P. Araújo et al.
Fig. 1. TG (a) DTG (b) curves of the films of starch/PE and additive with 1, 2 and 3 % of TEO.
polymeric films and Subbuvel et al. [44] when evaluating the matrix of
poly (lactic acid) films counting curcumin and fenugreek essential oil
obtained similar results.
3.5. Fourier transform infrared spectroscopy (FTIR) and principal
component analysis (PCA)
The FTIR spectra of the starch films with and without the addition of
TEO were very similar (Fig. 2a), the reason being well understood by the
analysis of the spectra of the TEO which has characteristic vibrational
bands similar to the bands of the starch films. The band presented be­
tween 3000 and 3500 cm− 1 corresponds to the OH stretch band in
glycerol, starch, and water in the film. It also exhibited a band at 2920
cm− 1 due to the CH fold, a band at 1640 cm− 1 attributed to the OH fold
of the water in the film, a band close to 1500 cm− 1 attributed to C–H
and a band at 1100 cm− 1 associated with C–O. Similar vibrational
spectra were found in starch films and they also show characteristic
signals of the vibrational spectra of pectic compounds [38].
The information obtained in the infrared, although it did not allow to
identify more subtle differences in relation to the spectra, was useful for
carrying out the PCA, which in turn showed the chemical differentiation
between the starch/PE film and the films added with TEO. In the graph
(Fig. 2b) it was possible to verify distinct regions that delimit the films
that had the addition of oil, represented by CS-1 % TEO, CS-2 % TEO and
CS-3 % TEO and the films without oil (CS).
The PC1 explained that the most relevant bands for the separation
between the starch films added and without TEO were around 1000 to
1200 cm− 1, referring to the characteristic C–O stretching of starch. This
band was very intense band in the spectra.
In both PC1 and PC2 (Fig. 2b), bands belonging to functional groups
characteristic of TEO, such as aromatic rings (C=C) at approximately
1600 cm− 1, the C– – O elongation at 1400 cm− 1 were responsible for the
shown separation. At approximately 600, 700, 800 and 900 cm− 1 the
presence of the C–H curvature was also responsible for the separation.
This proves the chemical differentiation obtained by the incorporation
of the TEO.
3.6. Transparency and colorimetry
The addition of TEO did not significantly alter the transparency of
the films, indicating uniformity between the formulations. Despite
studies reporting that the addition of essential oils in the polymer matrix
promoted an increase in light scattering, contributing to a reduction in
transparency. These results depend on the type and interaction of the
additive - polymer used [45]. Yeddes et al. [17] who did not observe
significant changes after the addition of rosemary essential oil to gelatin-
1356
International Journal of Biological Macromolecules 226 (2023) 1352–1359
chitosan-pectin films.
The L* value is an indication of the film’s brightness. This value
ranged from 94.66 to 93.88, which expresses that all films present
proximity to light color and loss of luminosity. However, it did not
impact the transparency of the films. The addition of oil generated a
slight decrease in this value and the result showed that there was no
statistically significant difference between the CS and CS-1%TEO sam­
ples. Evidencing that in small concentration, the oil did not significantly
alter the luminosity of the film. Li et al. [19] observed the same effect in
the evaluation of chitosan films added with TEO, whose addition of oil
promoted a reduction in luminosity.
The a* parameter represents the green (− ) and red (+) variation of
the films. This result was negative in all films, indicating the approxi­
mation with the green. Negative values were also found in CS films with
orange essential oil produced by Evangelho et al. [45] and in CS films
plasticized with glycerol and thymol manufactured by Nordin et al. [16].
In all the works cited, the tendency of CS films to be green in color was
evidenced.
The b* parameter, a component that reflects the variation of blue (− )
and yellow (+) color, increased considerably with the addition of oil,
indicating the gain of yellow color in the films. This can be seen by visual
analysis of the image of the films, shown in Table 2. This result comes
from the presence of curcumin in the chemical composition of the TEO,
since this component has a characteristic yellow-orange color. De
Campos et al. [46] evaluated that increasing curcumin in cassava starch
and poly (butylene adipate co-terephthalate)-PBAT films resulted in a
decrease in the L* luminosity parameter and a significant increase in the
b* value, indicating the trend of yellow color.
The total color difference obtained by calculating the ΔE* ranged
from 9.57 for the film containing 1 % TEO to 19.49 for the film con­
taining 3 % TEO. The increase in the oil content in the films generated a
greater color change, as a result of the increase in the a* and b* color
parameters.
3.7. Application of film on sliced bread
During the weekly analysis of the packages, the TEO migration
process was noticeable after the 4th week of analysis, due to the loss of
yellow color of the packages (Fig. 2S). In general, the breads packed by
films did not show any type of contamination during the nine weeks of
analysis. Meanwhile, the bread wrapped in the film of cellophane
showed signs of browning from the 6th week of analysis. The results
presented by the films produced in this study were satisfactory, since
they managed to inhibit the appearance of fungi during the 9 weeks of
analysis.
Thus, these results showed that it is possible to use TEO as an
M.N.P. Araújo et al.
Fig. 2. Medium infrared spectra (a) and Analysis of the main components (b) of the films of starch/PE and additive with 1, 2 and 3 % of TEO.
antimicrobial additive agent in the production of films for application in
active packaging. Mustapha et al. [47] explains that the antimicrobial
activity of TEO in the fungal species Aspergillus niger, one of the most
common fungi found in spoiled bread, was due to the presence of a high
percentage of Turmerone (35.46 %). As observed in the analysis of the
turmeric essential oil chromatography results of the present work, the
Turmerone compound was present (36.47 %), which was an indication
that this oil had action against food fungi.
Complementarily, analyzing the visible changes in sliced bread, as
performed by Priyadarshi et al. [29], only bread packed with the control
package showed signs of contamination. Fig. 3 shows the bread wrapped
in cellophane, identifying the browning points on the front (a) and
bottom (b). In view of the packages studied, the recommended one to
increase the shelf life of the breads was CS-3 % TEO, since the TEO at the
highest concentration studied increased the mechanical strength of the
films and did not cause undesirable changes in the thermal and barrier
properties.
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International Journal of Biological Macromolecules 226 (2023) 1352–1359
4. Conclusion
The addition of turmeric essential oil (TEO) in the corn starch film
was effective as food packaging to protect the sliced bread. The analysis
by principal components evidenced the chemical differentiation ob­
tained by the films added with TEO, proving the success of the incor­
poration of the oil in the polymeric matrix. TEO showed activity against
the bacterium Staphylococcus Aureus and satisfactory antioxidant activ­
ity due to the phenolic compounds content. The predominant active
chemical compounds were Turmerone, Ar – Turmerone and
β-Turmerone.
The CS-3%TEO film represented the best formulation developed. It
showed resistance, presenting twice the mechanical strength when
compared the CS film. The TEO provided a yellow color of the film,
characteristic of Curcumin, and reduced the flexibility of the film. On
the other hand, it kept the transparency, solubility, WVP and thermal
stability unchanged. The evaluation of the antifungal activity of the
M.N.P. Araújo et al.
Fig. 3. Bread packed with the control film. Front area (a) and lower side area (b).
films as sliced bread packaging resulted in a sample free of contamina­
tion during the 9 weeks. Thus, it can be concluded that the active film
developed with 3 % of TEO can be applied as food packaging, and it can
be recommended for the food industry as a biodegradable packaging
that is safe and able to extend the shelf life of sliced bread.
CRediT authorship contribution statement
M.N.P. Araújo, C.V.B. Grisi and C.R. Duarte were responsible for
Methodology, Data curation, Formal analysis, Writing – original
draft, Validation and Writing – review & editing; G.M. Vinhas and Y.
M.B. Almeida were responsible for Conceptualization, Project
Administration, Supervision, Funding Acquisition. All authors
contributed and approved the final draft of the manuscript.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgements
The authors would like to acknowledge the National Council for
Scientific and Technological Development (CNPQ, the Brazilian Agency)
for funding this project.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.ijbiomac.2022.11.248.
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