Food Packaging and Shelf Life 23 (2020) 100431

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Food Packaging and Shelf Life

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Packaging with cashew gum/gelatin/essential oil for bread: Release T

potential of the citral
Marília A. Oliveiraa, Maria L.C. Gonzagab, Maria S.R. Bastosa,*, Hilton C.R. Magalhãesa,
Selene D. Benevidesa, Roselayne F. Furtadoa, Rafael A. Zambellib, Deborah S. Garrutia
a
Embrapa Agroindústria Tropical, Rua Dra. Sara Mesquita 2270, CEP 60511-110, Fortaleza, CE, Brazil
b
Departamento de Engenharia de Alimentos, Universidade Federal do Ceará, CP 12200, 60455-760, Fortaleza, CE, Brazil

ARTICLE INFO ABSTRACT

Keywords: Cashew gum (CG) and gelatin (G) films were produced by incorporating ferulic acid (FA), as a cross-linking
Polysaccharide agent and Cymbopogon citratus essential oil (EO) for application bread packaging. The addition of FA and EO did
Cashew gum not significantly affect water vapor permeability (WVP) or opacity. There was a significant decrease in the
Gelatin solubility of the films, possibly due to matrix cross-linking. Morphology revealed films with smooth surfaces and
Ferulic acid
small cross-sections. Characteristic frequencies of CG and G demonstrated the cross-linking identified in infrared
Active film
(IV). EO cross-linked films showed lower rupture stress than non-EO films. Elongation-at-break increased from
2% (cross-linked) to 140% (cross-linked/EO). CGG/EO packaging was more permeable than commercial
packaging (polyethylene), giving bread more harshness. The experimental packaging provided six days of
preservation to the bread compared with three days for the commercial packaging. Neral and geranial presented
a higher proportion in the crust of the bread, suggesting antimicrobial action.

1. Introduction and essential oil. Ferulic acid, a powerful antioxidant extracted from
plants and grains, has been shown to be an excellent cross-linking agent
A wide variety of gums is produced in tropical and subtropical for the gell-gelatin polyelectrolyte complex, it acts on the amine groups
countries with attractive characteristics for commercial and industrial forming cross-links throughout the blended structure (Graf, 1992).
applications; a typical example is cashew gum, which has received at- Oxidation occurs employing 0.5% hydrogen peroxide solution. The acid
tention because it is similar to gum arabic, which is widely used in the forms a dimer as a side reaction or reacts with amine groups I and II
food industry. The Anacardium occidentale L. cashew tree is native to which are free in the gelatin chain to form OeN covalent bonds. The
northeastern Brazil, and exudes a gum in response to the attack of latter response can be reoxidized and linked to another amine, resulting
pathogens and lesions. In Brazil, cashew trees produce an average of in cross-linking.
700 g of cashew gum (CG) per year. Since there are more than 700 Research on active food packaging has been developed by in-
thousand hectares of planted cashew trees in Brazil, it is estimated that corporating substances that can alter the food environment, affecting its
the production potential of the CG is around 50,000 tons per year metabolism, providing greater protection, consequently improving
(Ribeiro et al., 2016), and it is continuously studied aiming towards quality and increasing its useful life (Bastarrachea, Dhawan, & Sablani,
different applications. Cashew gum exhibits stabilizing, emulsifying 2011; Zhao & Saldaña, 2019). Packaging is the last stage of production,
and adhesion properties in water, and can be molded into a film with and its choice is based on promoting the intrinsic quality of the product
the addition of a suitable plasticizer such as glycerol. However, even and its effectiveness in preserving and communicating this quality
with the use of glycerol in high concentrations, CG films are relatively (Licciardello et al., 2017).
fragile, inflexible and soluble, making it necessary to incorporate ge- The most commonly used polymers in packagings are derivated
latin and a cross-linking agent to improve the mechanical and solubility from petroleum products and not biodegradable, generating solid waste
properties. The need for cross-linking in packaging films is due to the from their final disposal and leading to serious environmental pro-
hydrophilicity of the materials, and is therefore a way to improve the blems. This opens the way for the use of natural biodegradable poly-
incorporation and adhesion of active substances such as antioxidants mers in manufacturing active food packaging (Arfat, Ejaz, Jacob, &

⁎
Corresponding author.
E-mail address: [email protected] (M.S.R. Bastos).

https://doi.org/10.1016/j.fpsl.2019.100431
Received 13 August 2019; Received in revised form 14 October 2019; Accepted 25 October 2019
Available online 18 November 2019
2214-2894/ © 2019 Elsevier Ltd. All rights reserved.
M.A. Oliveira, et al. Food Packaging and Shelf Life 23 (2020) 100431

Ahmed, 2017). of ± 0.01 mm. The thickness used was the arithmetic average of eight
Essential oils (EOs) extracted from plants and spices exhibit anti- measurements performed randomly along each evaluated sample.
microbial and antioxidant properties (Viuda-Martos et al., 2010), which
makes them excellent additives in the food industry as a substitute for 2.3.2. Opacity measurement
chemical additives (Alves-Silva et al., 2013). The composition of the oil Film samples were cut into a rectangle (8 x 40 mm) and placed on
and the specific interactions with the polymer determine its effective- the internal side of a spectrophotometer test cell (Agilent technologies
ness as an active ingredient (Chiralt & Atar, 2016). cary 60). The opacity was calculated using the following equation:
Cymbopogon citratus essential oil (EO) contains 40–80% of Citral,
Abs750
being widely used in the cosmetic, food and pharmaceutical industries Opacity =
X
(Martinazzo, Corrêa, Melo, & Barbosa, 2007) for its antifungal (de Sá
Silva et al., 2009) and antibacterial properties (Cimanga et al., 2002). Where: Abs750 is the absorbance value at 750 nm and X is the film
Therefore, the combined effect ferulic acid as cross-linked cashew thickness. The lower opacity values indicate higher sample transpar-
gum/gelatine films incorporated with commercial lemongrass ency (Farhan & Hani, 2017).
(Cymbopogom citratus) and on their physicochemical, microscopic,
spectrometric and mechanical characterizations and volatile com- 2.3.3. Water vapor permeability (WPV)
pounds of essential oil release was analyzed in this work to verify the This was determined in accordance with ASTM method E96-05
possibility of the an food packaging to bread. (ASTM, 2005). Eight replicate samples (60-mm diameter each) were
placed between the acrylic permeation cell fittings with a circular
2. Materials and methods opening (diameter = 50 mm), corresponding to the film area exposed
to migration. Each cell contained 5 mL of distilled water in its interior.
2.1. Cashew gum isolation The cells were placed in a desiccator (25 ± 5 °C temperature and
30 ± 5% relative humidity) containing silica gel. The weight of the
Gum exudate was collected from cashew trees from Embrapa cells was observed each hour on an analytical balance for 24 h.
Experimental Station at Pacajus (Fortaleza-Ceará). It was ground to 100
mesh particle size, and 100 g resulting powder was solubilized in 2.3.4. Solubility in water
300 mL water at the ratio of 1:3 (w/v) for 4 h, centrifuged at The water solubility determination was carried out on 3 cm x 3 cm
10,000 rpm at 4 °C for 20 min, and filtered to remove the insoluble film pieces based on the method proposed by Gontard, Duchez, Cuq,
materials. 300 mL exudate solution was add in 1200 mL commercial and Guilbert (1994) with some modifications. Previously dried and
ethanol at a 1:4 (v/v) ratio for 24 h to precipitate the polysaccharide. weighed samples were immersed in 50 mL of distilled water for 24 h at
The precipitate was washed with acetone, then vacuum filtered on 25 °C with stirring (75 rpm). The dry weight of the remaining film
funnel with sintered glass disc nº 1, this process being repeated for five pieces was obtained after filtration on previously dried and weighed
times and dried in a hot air circulation oven. This final powder is called filter paper and used to calculate the insoluble matter as a percentage of
“cashew gum” in this paper. Gelatin 225H B type (pH = 5.4) was kindly the initial dry weight (g/100 g). All the dry weights (of the initial and
supplied by Rousselot® (São Paulo, Brazil). final film pieces and the filter paper) were determined after drying at
105 °C for 24 h using a fan-assisted convection oven (Quimis model Q
2.2. Formulation of cross-linked film 31 4M22, Brazil).

Cashew gum (CG) and gelatin (G) at a weight ratio of 5.0CG:5.0 G 2.3.5. Contact angle
and 10% glycerol in relation to the total weight of the polymers were The contact angles of coating surface were measured on GBX -
added in water and kept under stirring for 4 h at 60 °C. Then 1% oxi- Digidrop MCAT (Modular Contact Angle Technology) system coupled to
dized ferulic acid (FA) was added relative to the total weight of the the CAN101 Tensiometer Optical Contact Angle Meter (KSV
polymers. After this period, the surfactants span 80 (S) at 0.6% and Instruments). Five drops of water were deformed on the surface of the
tween 80 (T) at 0.5% were added, reaching an Hydrophile-Lipophile films (FC and F1 to F4) individually. The angles were recorded by op-
Balance (HLB) of 10.8 and then 10% lemon grass essential oil (EO) tical system immediately after droplet deposition, ∼ 2 s, after 1 hour of
supplied by Embrapa was added. Homogenization occurred in an Ultra adhesion and after 2 hours with complete evaporation of water, all
Turrax T25 (IKA, Wilmington, NC, USA) for 10 min. The films were measurements being digitally determined. The assay was performed at
produced by the casting method and dried at room temperature. Four room temperature (∼ 25 °C).
films were prepared following the described procedure, called F1, F2,
F3 and F4. Film F1 contains all of the above components, F2 does not 2.3.6. Fourier transform infrared (FT-IR)
contain lemon grass essential oil, F3 does not contain surfactants and F4 The samples were analyzed as obtained by ATR/FT-IR in the
does not contain surfactants or essential oil. A non-crosslinked film medium infrared region, with wavelengths between 4000 and
without surfactants or essential oil was used as a control, called FC. The 400 cm−1 and 4 cm−1 resolution using the Perkin Elmer brand.
following were elaborated in order to facilitate identification of the
films: 2.3.7. Scanning electron microscopy (SEM)
F1(5.0CG:5.0 G:1.0 Gly:0.1FA:0.05T:0.06S:1.0EO), Morphological characterization was performed by scanning electron
F2(5.0CG:5.0 G:1.0 Gly:0.1FA:0.05T:0.06S), microscopy (SEM, Zeiss DSM, model 940A). Samples were coated with
F3(5.0CG:5.0 G:1.0 Gly:0.1FA:1.0EO), F4(5.0CG:5.0 G:1.0 Gly:0.1FA) platinum using a sputter coater (Electron Microscopy Sciences,
and Hatfield, PA, USA) and examined under SEM using an accelerating
FC (5.0CG:5.0 G:1.0 Gly). The commercial polyethylene film was voltage of 15 kV and a magnification of 3,000–6,000×.
obtained in the local market.
2.3.8. Mechanical properties
2.3. Film characterization Mechanical properties were tested according to ASTM Method
D882-09 (ASTM, 2009) on a mechanical tester (EMIC DL 3000) using
2.3.1. Thickness 12.8 cm film strips. The initial grip separation and cross-head speed
Film thickness was measured using a digital micrometer (Digimatic were set to 10 cm/min. Both force (N) and deformation (mm) were
model, Mitutoyo, Brazil) with a range of 0–25 mm and an accuracy recorded during extension. Tensile strength was calculated by dividing

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M.A. Oliveira, et al. Food Packaging and Shelf Life 23 (2020) 100431

the required force for film rupture by the cross-sectional area, and the films as barriers to water vapor bigger than for similar materials
elongation-at-break was calculated as the percentage increase in sample (Azeredo et al., 2012; Wu et al., 2016). The higher WVP values obtained
length. The elastic modulus was calculated from the slope of the for the FC, F3 and F4 films may be related to the absence of surfactants
stress–strain curve in the elastic deformation region. The reported va- in these materials, resulting in lower compaction of the structure by
lues are the averages of five measurements. decreasing of bonds. The higher thicknesses of these films may also
contribute to higher WVP values.
2.4. Film application as bread package Ferulic acid action was observed by Ou, Wang, Tang, Huang, and
Jackson (2005) and Silva, Santos, Coutinho, Mano, and Reis (2005),
2.4.1. Packaging preparation who reported no significant reduction in the vapor permeability of the
The package (Fig. 4c) was prepared using the F1 film, selected for its soy protein films cross-linked with this acid. In addition to noting the
mechanical resistance. Three 45 cm x 30 cm films each were closed by low influence of ferulic acid on the WVP of gelatin films. Cao, Fu, and
vacuum-sealed Selovac model 200B. The basic formulation bread He (2007) suggest that its small effect is due to the possible combina-
(wheat flour, margarine, baking powder, sugar and salt) was prepared tion of the hydroxyl groups of the acid with the water molecules, not
at the Laboratório de Cereais of Departamento de Tecnologia de Alimentos favoring permeation. Edible films made from sodium caseinate cross-
UFC - Brazil, cut into 1.25 cm thick x 15 cm in length, packed as shown linked with feluric acid were less permeable to water vapor than α-
in Fig. 4b and c and conditioned at room temperature (25 °C) for 8 days, tocopherol cross-linked films (Fabra, Hambleton, Talens, Debeaufort, &
during which the analyzes were performed. Chiralt, 2011). The presence of gelatin in the composition of the films
favors poor absorption of water since it can form organized structures,
2.4.2. Volatile compounds release in the bread with it being possible to recover part of the triple helix structure of the
Extraction of volatiles in the F1 films (the package) and packed collagen during the formation process of the film. The higher degree of
bread (Fig. 4c) was performed by Headspace Solid-Phase Microextrac- organization optimizes molecular packaging, resulting in films with
tion (SPME) using a carboxen/divinylbenzene/polydimethylsiloxane better gas barrier properties (Fan, Duquette, Dumont, & Simpson, 2018;
50/30 μm fiber. First, 0.5 g of sample film and bread separately were Yun & Dong, 2017). Benavides, Villalobos-Carvajal, and Reyes (2012)
cut in small pieces and weighed in 20 mL vials. The fiber was then formulated alginate films with oregano essential oil (EO) in varying
exposed into the headspace above the sample for 40 min, maintaining proportions and observed a porous structure, whose increase in EO
the temperature at 40 °C. The volatiles were thermally desorbed into content resulted in greater packaging of the layers, accompanied by an
the splitless injection mode of a gas chromatograph (GC Shimadzu improvement in water barrier properties. Ojagh, Rezaei, Razavi, and
GC2010 Plus, Japan) fitted with a 5% phenyl stationary phase capillary Hosseini (2010) incorporated cinnamon essential oil into chitosan films
column (30 m length, 0.25 mm id, 0.25 mm film thickness). Hydrogen and observed final films with leaf-like structures stacked in thick layers,
was used as the carrier gas at a flow rate of 1.0 mL min−1. The oven which presented better water barrier properties.
temperature was programmed from 70 °C to 180 °C at 4.0 °C min−1, and The solubility of the films after cross-linking decreased from 53 to
then raised to 250 °C at 8 °C min−1, held for 5 min. The injector tem- 3%, indicating good process efficiency. In a study by Cao et al. (2007),
perature was 250 °C and the FID detector was 260 °C. Gas Chromato- the effect of water on gelatin films cross-linked with ferulic acid was
graphy-Mass Spectrometry (GC–MS SHIMADZU QP2010, Japan) and classified as slight. The presence or absence of essential oils and sur-
Retention indices were used to identify neral and geranial. Analyses factants, associated or not, did not promote a change in solubility, as
were performed in triplicate. the data for F1 to F4 show.
Due to the similarity in the results obtained individually for the
2.4.3. Texture physicochemical properties analyzed herein, it can be shown that EO
The texture profile was obtained by the double compression test of was satisfactorily dispersed in the presence and absence of surfactants.
the bread slices packed in a TA-XT2i texture analyzer (Stable Micro The optical analysis of the films is essential to evaluate the accep-
Systems). A texture profile analysis (TPA) with velocity of 1.0 mm/s, tance of certain foods by the consumer. Transparency is essential for
compression distance of 40 mm, equivalent to 50% of compression, materials whose purpose as packaging is to promote good visualization
contact force of 5.0 g was performed using a SMS Aluminum Cylinder of the packaged product. The developed films are promising, since the
probe (P35). Texture Expert for Windows 1.20 program (Stable Micro non-cross-linked film (FC) did not register opacity and the reticulated
Systems) collected the data. The hardness parameter was evaluated in films presented small opacity values. It is important to note that such
this test. behavior was independent of the composition and thickness of the
films. For thickness, the fluctuation in the values is due to the mold
2.5. Statistical analyses casting method, a fact reported by other authors (Cao et al., 2007;
Henrique, Cereda, & Sarmento, 2008).
The volatile compounds data were evaluated by regression analysis, The results (Table 1) show similar values for crosslinked films (F1 to
while the quality of the models (linear or quadratic) was evaluated by F4), indicating that the presence or absence of surfactants, essential oil
the coefficient of determination (R2) and by the lack of fit test. The film and glycerol did not influence the adhesive properties of the films
characterization data were submitted to analysis of variance (ANOVA) structural composed by gelatin cashew gum. Non-crosslinking of the FC
and Tukey test with a level of 5% of significance for comparison of film, which gives it greater solubility, may account for its lower contact
means. All analyses were performed using using the XLSTAT v. 18.1 angle value. The low solubility of films (F1 to F4) may be related to the
statistical software program (Addinsoft). branched molecular structure of cashew gum (Ribeiro et al., 2016). This
characteristic added to the presence of cross-linking may contribute to
3. Results and discussion the non-exposure of hydroxyls present in glycosidic units, not favoring
the formation of hydrogen bridges with water and, consequently, their
3.1. Characterization of films solubility. Fig. 1 illustrates the behavior of sessile drop on the surface of
the films. The interval recording of 2 h shows the efficient adhesion of
3.1.1. Thickness, opacity, WVP, solubility and contact angle water to film surfaces, regardless of their compositions. After 1 h of
Table 1 presents the properties of thickness, opacity, water vapor contact, it was observed that the drop deformed the films, without
permeability, solubility and contact angle of films obtained under dif- breaking them and spreading the droplets on the surfaces. This may be
ferent conditions. As shown in Table 1, cross-linking through ferulic related to the ability of the material to absorb water by capillary in-
acid did not influence the WVP of the films (F1 to F4). The results show filtration or by reorienting hydrophilic groups towards the surface as a

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M.A. Oliveira, et al. Food Packaging and Shelf Life 23 (2020) 100431

Table 1
Thickness, water vapor permeability, solubility and opacity for F1, F2, F3, F4 and FC films.
Film Thickness§ WVP Solubility Thickness† Opacity Contact angle
(μm) (g.mm.k. Pa−1. h−1. m-2) (%) (μm)

FC 59.67 ± 1.50bc 1.42 ± 0.20 c

53.40 ± 2.50a 76.59 ± 3.27b 0.00 ± 0.00 d
50.98 ± 2.23 a

F1 58.94 ± 3.93 bc 1.78 ± 0.07 bc

3.04 ± 0.31 b 47.21 ± 5.94 c 0.11 ± 0.00 a
61.66 ± 1.30 b

F2 49.90 ± 6.25 c 1.72 ± 0.40 bc

2.39 ± 0.42 b 51.10 ± 7.10 c 0.10 ± 0.00 c
61.78 ± 1.78 b

F3 95.10 ± 8.42 a 2.60 ± 0.30 a

3.00 ± 0.48 b 58.13 ± 10.66 c 0.10 ± 0.00 bc
62.72 ± 2.91 b

F4 68.90 ± 5.61 b 2.10 ± 0.22 ab

3.03 ± 0.35 b 46.35 ± 6.27 c 0.11 ± 0.00 ab
60.68 ± 1.25 b

§
Thickness for WVP (mean of eight replicate samples, each measured eight times).
†
Opacity thickness (average of six replicate samples, each measured eight times).
Mean of three values with standard deviation, the same letter in the row indicates that there is no significant difference between the means by the Tukey test
(p < 0.05).
F1(5.0CG:5.0 G:1.0 Gly:0.1FA:0.05T:0.06S:1.0EO), F2(5.0CG:5.0 G:1.0 Gly:0.1FA:0.05T:0.06S), F3(5.0CG:5.0 G:1.0 Gly:0.1FA:1.0EO), F4 (5.0CG:5.0 G:1.0 Gly:0.1FA) and FC
(5.0CG:5.0 G:1.0 Gly).

function of hydration generated by interaction with water (Gugliuzza & stretches of the C–H bonds of CH3 and C]O of the aldehyde, are added
Drioli, 2004). The non-rupture of the films was better recorded after 2 h to those recorded at 2923 cm−1 and 1633-1638 cm−1, respectively. As
of application, with drop evaporation. proposed in this work, the bond that structures the cross-linking is
likely made between the amino group (NH2) of the gelatin and the
hydroxyl group (OH) of ferulic acid, the registration of which can be
3.1.2. Fourier transform infrared (FT-IR)
identified by symmetrical stretching at 1546 cm−1 for NO (Silverstein
The IR spectra of the films, ferulic acid, cashew gum, and gelatin are
et al., 2007), present in all cross-linked films.
shown in Fig. 2. Characteristic frequencies of the present polymers, CG
and G and FA on the films are reported at 3295-3304 cm−1 for OH, and
NH stretches, 2923 cm−1 for asymmetric stretches CH of CH2, 3.1.3. Scanning electron microscopy (SEM)
1150 cm−1 for CO (anomeric carbon), 1072-1077 cm−1 for COC (gly- Fig. 3 micrographs were obtained for the control-FC and F3-sample
cosidic bonding), 1031-1036 cm−1 for alcohol CO, with the latter three films. The F1, F2 and F4 images were suppressed by F3 similarity, de-
being specific for CG. C]O acid stretches, characteristic of gelatin and spite differences in compositions. The FCa and F3a films show homo-
ferulic acid, are recorded at 1650 and 1679 cm−1, respectively. The geneity in the surfaces with no irregularities, as reported by Cao et al.
record of this group in the films can be observed displaced in the range (2007). The recording of air bubbles is due to the casting method used
of 1633-1638 cm−1. in the molding of films (Mathew & Abraham, 2008).
The presence of ferulic acid on the films is also recorded in the The transverse sessions (FCb and F3b) show higher compression in
stretches at 1454 cm−1, the characteristic frequency of C]C aromatic F3b, with few pores without depth. The bumps in some points are due
ring bonds (Wang, Sun, Liu, Li, & Ma, 2011; Yeung, Eskici, & Axelsen, to the blade action used in the cut. The compact aspect observed may be
2013; Silverstein, Webster & Kiemle, (2007)). Three intense bands at related to the degree of interaction between the gelatin molecules, one
1618, 1598 and 1518 cm-1 observed in the aromatic ring of fungal acid of the structural polymers of the film, provided by the cross-linker ac-
appear in some regions between 1630 to 1556 cm-1. tion. Mathew and Abraham (2008)) observed a similar compact aspect
EO-specific frequencies added in F1 and F3, such as the asymmetric for composite chitosan and starch films cross-linked with ferulic acid.

Fig. 1. Wettability behavior of FC and F1-F4 films after 2 s, 1 h and 2 h.

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M.A. Oliveira, et al. Food Packaging and Shelf Life 23 (2020) 100431

Fig. 2. Infrared spectra of the films samples (F1, F2, F3, and F4) and control (FC), Ferulic acid (FA), Gelatin (G) and Cashew gum (CG).

Considering the similarity in the compositions of FC and F4, the ab- on the concentration of the cross-linking agent, the literature indicates
sence of ferulic acid (the only non-common component) may respond to that the cross-linking process makes the polymer matrix structure more
the aggregates visualized in FCb. The presence of the essential oil did rigid, leaving the films less flexible (Dammak, Bittante, Lourenço, & do
not promote morphological changes in the F1 and F3 films when Amaral Sobral, 2017). The obtained results reveal a good cross-linking/
compared to the other cross-linked F2 and F4 films. matrix ratio able to flexibilization the polymeric chain. The E in the
cross-linked films decreases and the ε increases considerably, because in
the case of ferulic acid, the cross-linker can preserve the matrix struc-
3.1.4. Mechanical properties ture in its action, herein formed by CG:G, resulting in a more flexible
The mechanical properties of the film indicate the expected in- film with lower tensile strength, especially in F1 and F3, which present
tegrity under stress conditions that would occur during processing, EO.
handling, and storage. The stiffness, strength, and flexibility of the film A decrease in tensile strength caused by adding EO in films based on
are described by Young’s modulus (E), tensile strength (σ) and elon- polysaccharides was repeatedly observed (Atarés, Pérez-Masiá, &
gation-at-break (ε). Table 2 shows the modulus, tensile and elongation Chiralt, 2011; Ma, Hu, Wang, & Wang, 2016; Sánchez-González, Cháfer,
values for the ferulic acid cross-linked films with and without the sur- Chiralt, & González-Martínez, 2010; Sánchez-González, Chiralt,
factants, lemongrass essential oil, as well as the non-cross-linked (FC) González-Martínez, & Cháfer, 2011). Tongnuanchan, Benjakul, and
film. It is observed that FC showed to be more rigid than the cross- Prodpran (2012) and Limpisophon, Tanaka, and Osako (2010) observed
linked films. However, its elongation-at-break was very low. Depending

Fig. 3. Micrographs of the surfaces (a) and cross-sections (b) of the films: FCa and FCb (5.0CG: 5.0 G: 1.0 Gly); F3a and F3b (5.0CG: 5.0 G: 1.0 Gly: 0.1FA: 1.0EO).

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M.A. Oliveira, et al. Food Packaging and Shelf Life 23 (2020) 100431

Table 2
Mechanical properties for the F1, F2, F3, F4 and control (FC) films.
Films Thickness(μm) E(Mpa) σ(MPa) ε(%)

b a a
FC 74.00 ± 5.00 3.54 ± 1.34 58.36 ± 0.03 2.97 ± 0.98 b
F1 51.53 ± 0.79 cd 0.59 ± 0.25 b
19.16 ± 4.18 c
141.57 ± 46.75 a

F2 46.12 ± 0.92 d 0.09 ± 0.00 c

46.60 ± 1.25 b
105.50 ± 65.42 a

F3 140.60 ± 5.16 a 0.27 ± 0.09 c

19.40 ± 0.65 c
121.69 ± 41.75 a

F4 56.46 ± 1.49 c 0.14 ± 0.67 c

50.51 ± 2.18 ab
137.13 ± 34.89 a

Mean of three values with standard deviation, same letter in the line indicates that there is no significant difference between the means by Tukey test (p < 0.05).

Fig. 4. (a) Neral and geranial determination in

percentage of area and their respective equa-
tions. Bread crust: neral (□): y = 176.37x² –
731.18x + 2057.9 R² = 0.9309; geranial (●):
y = 212.11x² – 929.76x + 2278.2
R² = 0.9451 and bread crumbs: neral (Δ) :
y = 28.111x² – 93.522x + 340.27
R² = 0.7308; geranial (▼) : y = 22.114x² –
73.734x + 233.45 R² = 0.6725 (b) Bread in
commercial packaging (c) Bread in CGG / EO
film and (d) aspect of bread wrapped in CGG /
EO film over days.

that the incorporation of citric oil resulted in a reduction in the tensile Farahmandghavi (2015) to justify the reduction of tensile strength in
strength of edible films of fish skin gelatine, which was attributed to the gelatine-chitosan fish films added with Origanum vulgare essential oil.
lower interaction between the gelatin molecule and provides flexible Therefore, in the films studied herein greater flexibility and lower
domains within the film. The effects of destabilization phenomena stiffness of the cross-linked films is a probable consequence of the
during the film drying were considered by Hosseini, Rezaei, Zandi, and presence of the essential oil, favoring the behavior of the glycerol in

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M.A. Oliveira, et al. Food Packaging and Shelf Life 23 (2020) 100431

study (Fig. 4d).

The efficiency of active packaging by the action of essential oils was
verified by Priyadarshi, Sauraj Kumar, Deeba, Kulshreshtha, and Negi
(2018) by incorporating apricot kernel oil into chitosan films. These
authors not only observed better antioxidant activity, but also sub-
stantial antibacterial activity against Gram-negative E. coli, as well as
Gram-positive B. subtilis bacteria. The films were also effective in in-
hibiting mold growth on bread for ten days, thus increasing its shelf life.
Antioxidant action was also reported by the authors, similar to that
disclosed for chitosan films added with green tea extract as presented
by Siripatrawan and Harte (2010). Similarly, by adding cinnamalde-
hyde essential oils to wheat glycine films, Balaguer, Lopez-carballo,
Catala, Gavara, and Hernandez-munoz (2013) obtained a material with
antibacterial and antifungal action, which promoted a drastic reduction
of fungi, and consequently extended the shelf life of the bread and
packaged cheese.
Fig. 5. Bread hardness packed in CGG / EO film and commercial packaging.
An important issue nowadays is the selection of packaging systems
which are not only efficient, meaning capable of maintaining quality
leaving spaces in the matrix structure. The action of the essential oil characteristics, but also useful in containing the environmental impact
was also observed in films based on synthetic polymers of cellulose and costs generated by the production and disposal of packaging
acetate, with reduction of the tension at rupture by the action of (Licciardello et al., 2017). In this aspect, developed packaging is fa-
Cymbopogon citratus (Oliveira et al., 2017). vorable, since all the components are biodegradable (Biswas et al.,
The action of EO on elongation-at-break of films was also reported. 2018) and of affirmative action in the biochemical processes of the
Pranoto, Salokhe, and Rakshit (2005) observed that the proportion of environment.
EO determines its effect on elongation-at-break. Shojaee-aliabadi,
Hosseini, and Amin (2013) found an increase in the elongation of κ-
3.1.6. Texture
carrageenan film caused by adding Satureja hortensis EO. Zinoviadou,
Fig. 5 shows hardness data for bread packed with CGG/EO film and
Koutsoumanis, and Biliaderis (2009) incorporated oregano EO into
polyethylene (PE) film (commercial packaging). It is observed that the
serum protein films, and observed an increase in the elongation prop-
hardness is inversely proportional to the water levels of the bread.
erties at an oil concentration of up to 1%; a fact explained by the de-
Bread packed in CGG/EO film is more stringent than food wrapped in
velopment of composite film with discontinuities.
commercial packaging of PE over the course of days because the WVP
Different behaviors have also been observed. Increased tensile
of the CGG/EO film (1.78 g.mm.k.pa−1. h−1. m-2) is high enough to let
strength by adding EO in films was observed by Atarés, Bonilla, and
the bread lose its moisture, making it harder as the days go by com-
Chiralt (2010) in cinnamon EO soy protein films, and by Ojagh et al.
pared to polyethylene packaging (0.008 g.mm.k.pa−1. h−1. m-2), al-
(2010) who incorporated the same oil into chitosan films. These studies
ready widely used in the market. Mudgil, Barak, and Khatkar (2016)
justified such behavior by the strong interaction between the EO and
observed that the bread hardness remained almost unchanged up to
the polymer matrix, promoting chain cross-linking. Ojagh et al. (2010)
60% water level, but the bread hardness decreased with the increasing
also found a decrease in elongation-at-break in films, similar to Moradi
water level beyond 60%. The increase in permeability to water vapor
et al. (2012) when incorporating Zataria multiflora EO into chitosan
(WVP) and water absorption is one of the main factors that may con-
films, where the latter attributed this to an increase in the pore size of
tribute to the degradation of packaging made with CGG film (Oliveira
the films, creating possible rupture points.
et al., 2018). From the characteristics presented, the packaging under
study proved to be efficient in protecting the bread from microbial
3.1.5. Application of the film as bread packaging action for at least six days, due the less water available for the microbial
Several studies have demonstrated the efficacy of packaging using growth as demonstrated in Fig. 4d
various materials and techniques in maintaining bread quality char-
acteristics, delaying moisture loss and fungus growth (Latou, Mexis,
Badeka, & Kontominas, 2010; Licciardello, Cipri, & Muratore, 2014; 4. Conclusion
Pagani, Lucisano, Mariotti, & Limbo, 2006; Piergiovanni & Fava, 1997;
Cozmuta et al., 2015). These results imply in an increase in the shelf life Ferulic acid was efficient in cross-linking the GC:G matrix, pro-
of the bread. viding films with low solubility and opacity, and without significant
Research of the active components identified neral and geranial in change in water vapor permeability when compared to FC. The IR
higher proportions in the bread crust, and increasing throughout the spectrum confirmed the cross-linking at 1546 cm−1 relative to the N–O
days (Fig. 4a), with data indicating significant differences by the value covalent bond. The cross-linking provided more compact films. The
of P < 0.0001. Considering the arrangement of the slices inside the cross-linked films were shown to be less rigid according to the Young’s
package (Fig. 4b Commercial packaging and 4c CGG/EO packaging) modulus and having higher elongation-at-break by more than 100%,
and the observed shelf life, the antimicrobial barrier action do film was which gives them a plastic material characteristic. The essential oil
observed mainly in the bread crust. This fact can be suggested by re- favored the mechanical properties, with a 38% decrease in the tensile
taining the active compounds of the Cymbopogon citratus essential oil strength in the films. In general, the presence and absence associated or
(EO) incorporated in the experimental film. The presence of the assets not of the surfactants and essential oil did not affect the investigated
in the core is added to the bread’s stability. Such behavior is relevant characteristics. The essential oil promoted bread preservation. The re-
due to the absence of preservatives in the food used. sults present the films as an alternative to food packaging.
Assay research was completed after the sixth day of storage due to
fungus presence (visual observation) (Fig. 4c). This fact reveals oxygen Declaration of Competing Interest
permeability of the packaging under study, however without compro-
mising its effectiveness given that the satisfactory storage period pro- The authors declare that they have no conflict of interest regarding
moted six days in function of the commercial package evaluated in this this manuscript.

7
M.A. Oliveira, et al. Food Packaging and Shelf Life 23 (2020) 100431

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