Food Chemistry 269 (2018) 142–149

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Butylated hydroxyanisole encapsulated in gelatin fiber mats: Volatile release T

kinetics, functional effectiveness and application to strawberry preservation
⁎ ⁎
Linlin Lia, Hualin Wanga,c, , Minmin Chena, Suwei Jianga, Shaotong Jiangb,c, Xingjiang Lib, ,
Qiaoyun Wangb
a
School of Chemistry and Chemical Engineering, Hefei University of Technology, 230009 Hefei, Anhui, PR China
b
School of Food Science and Engineering, Hefei University of Technology, 230009 Hefei, Anhui, PR China
c
Anhui Institute of Agro-Products Intensive Processing Technology, 230009 Hefei, Anhui, PR China

A R T I C LE I N FO A B S T R A C T

Keywords: Butylated hydroxyanisole (BHA) encapsulated in gelatin (GA) (GA-BHA) fiber mats were fabricated via elec-
Electrospinning trospinning technique and applied to strawberry preservation. The volatile release kinetics and functional ef-
Butylated hydroxyanisole fectiveness of the mats were investigated. BHA was high efficiently encapsulated in GA fibers and the antioxidant
Gelatin activity of BHA could be well protected. The encapsulation of BHA enhanced the stability of GA and favored
Strawberry
structure transition of GA from random coil and β-turns to α-helix and β-sheet. The GA-BHA mats showed good
antibacterial activity against Staphylococcus aureus, and the predominant volatile release mechanism of BHA
from mats was Fickian diffusion. Furthermore, the mats also showed broad-spectrum antifungal activity against
four mould genera (Rhizopus sp., Mucor sp., Aspergillus sp. and Penicillium sp.). The shelf-life of strawberry can be
prolonged effectively in the presence of GA-BHA mats during storage. Results suggested that the GA-BHA mats
may have a great potential in active food packaging.

1. Introduction efficient technique for the encapsulation of bioactive entities (Neo

Active food packaging offers new opportunities for food preserva-
tion (Etxabide, Uranga, Guerrero, & de la Caba, 2017; Wen, Wen, Zong,
Linhardt, & Wu, 2017). Antimicrobial packaging is one of the advanced
food packaging technologies and is extensively studied for the appli-
cation to food preservation. Phenolic compounds (Rui et al., 2017),
flavonoids (Aytac, Kusku, Durgun, & Uyar, 2016), bacteriocins
(Balciunas, Castillo Martinez, Todorov, Franco, Converti, & Oliveira,
2013), and antimicrobial enzymes (Wang, Yue, & Lee, 2015) are the
commonly used entities in antimicrobial packaging. Among them,
phenolic compounds have attracted more attention because of their
better antibacterial and antioxidant activities (Franco, Galeano-Díaz
et al., 2014; Zhang et al., 2014). Butyrate hydroxyanisole (BHA) is a
commonly used food antioxidant and antiseptic due to its low cost and
availability (Wu et al., 2015; Zhao & Hao, 2013). In order to expand the
application of BHA, we evaluated the application of BHA to preserva-
tion of strawberries in the present work. As BHA is a volatile compound
which is easily oxidized, an effective encapsulation of BHA is needed for
the purpose of its control release. However, less relative research work
has been done with BHA.
Electrospinning has been widely used as a simple, versatile and
et al., 2013; Wen et al., 2016). In comparison with the traditional films
prepared by casting or coating methods, electrospun fibers are more
responsive to surrounding atmosphere change and enable the con-
trollable release of encapsulation entities due to its larger specific sur-
face area and higher porosity (Vega-Lugo & Lim, 2009). Gelatin is a
kind of soluble peptidic compound derived from collagen (Gómez-
Guillén, Giménez, López-Caballero, & Montero, 2011), which possesses
good fiber-forming capacity besides excellent biocompatibility, non-
toxicity, abundance and low cost (Torkamani, Syahariza, Norziah, Wan,
& Juliano, 2018). Some bioactive entities such as asiaticoside
(Sikareepaisan, Suksamrarn, & Supaphol, 2008), vitamin A and E (Li
et al., 2016) and curcumin (Deng, Kang, Liu, Feng, & Zhang, 2017) were
successfully encapsulated in gelatin matrix via electrospinning tech-
nique.
In comparison with non-volatile bioactive compounds, the volatile
ones encapsulated in packaging matrix can be easily released and vo-
latilize to the limited storage space of food, exerting antibacterial ac-
tivity towards microorganism or antioxidant towards lipids and protein.
There are a few reports regarding the development and application by
utilizing the volatility of antibacterial compounds (Avila-Sosa et al.,
2012; Shojaee-Aliabadi et al., 2013). Herein, BHA encapsulated in

⁎
Corresponding authors at: School of Chemistry and Chemical Engineering, Hefei University of Technology, 230009 Hefei, Anhui, PR China (H. Wang).
E-mail addresses: hlwang@hfut.edu.cn (H. Wang), lixingjiang1978@hfut.edu.cn (X. Li).

https://doi.org/10.1016/j.foodchem.2018.06.150
Received 13 March 2018; Received in revised form 26 June 2018; Accepted 30 June 2018
Available online 02 July 2018
0308-8146/ © 2018 Elsevier Ltd. All rights reserved.
L. Li et al. Food Chemistry 269 (2018) 142–149

Fig. 1. Schematic illustration of BHA volatile release and application to strawberries.

Fig. 2. (A) FTIR spectra of BHA powders, GA and GA-BHA fiber mats, (B) Hydrogen bond between GA and BHA, (C) CD spectra of GA-BHA solutions at 25 °C, and (D)
DSC curves of GA and GA-BHA fiber mats.

Table 1 2. Materials and methods

Secondary structure proportions of GA obtained from deconvoluted CD spectra.
α-helix (%) β-sheet (%) β-turns (%) Unordered (%) 2.1. Materials

GA 27.2 ± 0.6a 7.0 ± 0.2a 18.9 ± 0.7a 48.2 ± 1.1a Porcine gelatin (from porcine skin, G2500, type A) with moisture
GA-BHA20 28.9 ± 0.7a 7.3 ± 0.3a 16.1 ± 0.5b 47.5 ± 0.9a
content no higher than 15% was bought from Sigma (St Louis, MO,
GA-BHA30 29.1 ± 1.1a 11.0 ± 0.5b 13.2 ± 0.4c 46.9 ± 0.4a
GA-BHA40 31.9 ± 0.9b 23.1 ± 0.5c 13.0 ± 0.3c 31.8 ± 0.5b
USA). Hydrochloric acid (HCl), sodium chloride, sodium bicarbonate
and sodium carbonate were supplied from Sinopharm (Shanghai,
Data with the same superscript letter in the same column indicate that they are China). Glacial acetic acid (> 99%), butylated hydroxyanisole
not statistically different (p > 0.05). The data (mean ± SD) are results from (BHA, > 99%), and ethyl alcohol (> 99%) were provided by Aladdin
three independent experiments. (Shanghai, China). Folin-Ciocalteu’s reagent, phosphate buffered solu-
tion (PBS, 1X) was obtained from Beijing Solarbio Science &
gelatin fiber mats were fabricated via electrospinning technique and Technology Co., Ltd (Beijing, China). Beef extract and peptone were
applied to strawberry preservation. Attentions were focused on the bought from Sangon Biotech Co., Ltd (Shanghai, China). Strawberries
volatile release kinetics in limited space and functional effectiveness (Fragaria × ananassa Akihime) were purchased from commercial
(antioxidant, antibacterial and antifungal activities) of the mats. greenhouses in a local farm (Hefei, Anhui) in November. All fruits
without additional pretreatment were selected based on the same ma-
turity (> 75% red surface color), no physical damage and uniform size.
Milli-Q water from a Millipore purification system (Bedford, MA, USA)

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L. Li et al. Food Chemistry 269 (2018) 142–149

Fig. 3. (A) Cyclic voltammograms of GA-BHA30 fiber mats at different release times, (B) Released amounts of BHA from fiber mats to the space, (C) Cyclic
voltammograms of BHA oxidation at various scan rates, (D) Dependence of ip on the v1/2 for GA-BHA30, (E) Growth curves of S. aureus exposed to fiber mats, and (F)
Inhibition efficiency curves of GA-BHA fiber mats against S. aureus.

was used in experimental process. As a kind of common food-borne
pathogenic bacteria, Staphylococcus aureus (S. aureus, ATCC 6538) from
the China Center of Industrial Culture Collection (Beijing, China) was
selected as model testing bacteria for the antibacterial evaluation of
fiber mats.
2.2. Preparation of fiber mats
Gelatin (GA) solutions (30%, g/ml) were prepared by completely
dissolving GA particles (4.8 g) in a mixture medium (16 ml, acetic acid/
ethanol/water = 3/2/3, w/w/w) at 25 °C under magnetic stirring for
5 h. After then, the requisite BHA was added into the medium to pre-
pare a serial of electrospinning solutions (20, 30 and 40%, i.e. 0.96,
1.44 and 1.92 g, on basis of GA weight) and the relative electrospinning
fibers were named GA-BHA20, GA-BHA30 and GA-BHA40, respec-
tively.
All the fiber mats were fabricated at 25 °C by an electrospinning
device equipped with a high voltage power supply (DW-P403-1ACCC,
Tianjin, China), a needle with an internal diameter of 0.6 mm, a syr-
inge, a ground electrode and a copper sheet. Subsequently, the fiber
mats were dried in an oven (25 °C, 8 h) to weight lose less than 0.05%
per 2 h and then sealed preservation. According to our preliminary
experimental data, the electrospinning parameters were determined as
follows: relative humidity (50%), supplied voltage (12 kV) and distance
(13 cm) between tip and collector.

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L. Li et al. Food Chemistry 269 (2018) 142–149

Table 2 ca
EE (%) = × 100
Correlation coefficients (R2) of the different models and the release exponents ct (1)
(n) of the Ritger and Peppas model.
where ca and ct represent the actual and theoretical concentrations
model R2 release kinetic transport of BHA (mg/ml), respectively.
data mechanism

GA-BHA20 Ritger and 0.9345 k = 0.1775 Fickian diffusion 2.3.4. Cyclic voltammetry
Peppas Cyclic voltammetry (CV) was performed at 25 °C on a CHI 660D
n = 0.43 potentiostat (Shanghai, China) equipped with a working electrode
Zero-order 0.7931 k = 0.0120
First-order 0.9967 k = 0.0672 Fickian diffusion
(glassy carbon disk with 3 mm in diameter, MF-2012), a reference
Higuchi 0.9233 k = 0.1285 electrode (BAS Ag/AgCl, +207 mV vs SHE), and a counter electrode
Hixson-Crowell 0.9715 k = 0.0125 Fickian diffusion (platinum wire). The cyclic voltammograms were recorded by scanning
GA-BHA30 Ritger and 0.9050 k = 0.2804 Fickian diffusion between −1.0 and 1.0 V.
Peppas
n = 0.32
2.3.5. BHA volatile release
Zero-order 0.6988 k = 0.0100
First-order 0.9932 k = 0.1122 Fickian diffusion The investigation of the volatile release behaviors of BHA from the
Higuchi 0.8569 k = 0.1077 GA-BHA fiber mats were performed at 37 °C within 72 h in well-sealed
Hixson-Crowell 0.9307 k = 0.0118 Fickian diffusion sterile glass chambers (21 × 10.5 × 5 cm, length × width × height),
GA-BHA40 Ritger and 0.8423 k = 0.4120 Fickian diffusion which were placed in a SPX-150B-Z biochemical incubator (Shanghai,
Peppas China). Fiber mats (6 × 6 cm) with a weight of 75 mg were placed in
n = 0.23 each of the nine sterile glass chambers and sealed well, respectively,
Zero-order 0.5683 k = 0.0076
and then subjected to release testing. At fixed time, the specimen was
First-order 0.9954 k = 0.1883 Fickian diffusion
Higuchi 0.7462 k = 0.0844 successively taken out from the each chamber and then completely
Hixson-Crowell 0.8891 k = 0.0166 Fickian diffusion dissolved in 50 ml electrolyte solution (PBS/ethanol = 7/3, v/v,
pH = 1 by HCl) at 25 °C with gentle stirring before being subjected to
the CV testing at a scanning rate of 0.1 V/s.
2.3. Methods According to the cyclic voltammograms of BHA in 50 ml electrolyte
solutions (PBS/ethanol = 7/3, v/v, pH = 1 by HCl) (Fig. S2A), the BHA
2.3.1. Viscosity and conductivity of electrospinning solutions standard curve (Fig. S2B) was established by plotting anodic oxidation
For the evaluation of the influence of BHA on the morphology of peak total charge (Qt) vs BHA concentration (CBHA). The released
fibers, a rotational viscometer (NDJ-5S, Shanghai, China) and an elec- amount of BHA to the space was determined by calculating the residual
tric conductivity meter (DDL-80, Shanghai, China) were used to mea- BHA in the fibers using the regression equation (Fig. S2B, in set:
sure the viscosity and conductivity of the solutions (25 °C), respectively. y = 0.1947x + 0.1652, R2 = 0.9991), where x is CBHA (mM) and y is Qt
(mC/cm2).
2.3.2. Morphology and structure of GA-BHA fibers To investigate the mass volatile mechanisms of BHA from fiber
A SU8020 scanning electron microscopy (SEM, Hitachi, Japan) was mats, the release profiles were described by Ritger and Peppas model as
used to observe the fiber morphologies, specimens sputter-coated with follows (Ritger & Peppas, 1987a, 1987b):
gold. The average diameter of fibers was determined by 100 randomly Mt
selected counts in the images. The interaction between GA and BHA in = ktn
M∞ (2)
the fibers was investigated by Fourier transform-infrared (FT-IR)
3 3
spectra on a Nicolet 6700 spectrometer (Thermo Nicolet, Madison, WI, where Mt (mg/cm ) and M∞ (mg/cm ) represent the release amount
USA) in the range 4000–500 cm−1. The effects of BHA on the secondary of BHA at time t and equilibrium, respectively, k the release rate con-
structure of GA was investigated by circular dichroism (CD), which stant, and n the release exponent indicating the release mechanism. In
were recorded on a spectropolarimeter (JASCO J1500, Tokyo, Japan) at addition, four following kinetic models were also employed to further
M
25 °C in a quartz cell (1.0 mm) with wavelength range of 190–250 nm at analyze release characteristics: M t = kt (Zero-order model),
∞
a scan rate of 50 nm/min (Liu et al., 2017). The GA concentration of M
1− M t = e - kt (First-order model),
Mt
= kt1/2 (Higuchi model) and
M∞
each testing specimen was of 0.2 mg/ml by dissolving fiber mat in ∞
1/3
ethanol aqueous (20%, w/w). According to the CD spectra, the sec- (1− )
Mt
M∞
= −kt (Hixson-Crowell model).
ondary structure proportions of GA were calculated by CD Pro software
using CDSSTR, CONTIN, and SELCON3 methods and each result was 2.3.6. Antioxidant activity of GA-BHA fiber mats
the average of the three methods. Differential scanning calorimetry The fiber mats with weight of 75 mg were completely dissolved in
(DSC) was performed on a DSC Q2000 (TA Instruments, New Castle, 50 ml electrolyte solution (PBS/ethanol = 7/3, v/v, pH = 1 by HCl) at
USA). The mats (about 3 mg) were tested from 25 to 250 °C at a rate of 25 °C with gentle stirring, and then the solution was subjected to CV
10 °C/min (50 ml/min, nitrogen atmosphere). testing at six scanning rates (0.01–0.36 V/s).

2.3.3. Encapsulation efficiency of fibers
Encapsulation efficiency (EE) of GA-BHA fiber mats were de-
termined according to the Folin-Ciocalteu method with some mod-
ification (Rebelo, Rego, Ferreira, & Oliveira, 2013). The mats with a
weight of 0.01 g were washed with 20 ml ethanol for the removal of the
BHA on fibers surface before dissolving in ethanol aqueous (50 ml, 20%
w/w). The absorbance (A) of resulting solutions was detected at 765 nm
with a UV/Vis-754 PC spectrophotometer (Shanghai, China). The actual
concentrations of BHA were determined on the basis of the obtained
regression equation as shown in the inset of Fig. S1, and the EE of the
fiber mats was calculated as follows:
2.3.7. Antibacterial activity of GA-BHA fiber mats
The antibacterial activity of fiber mats against S. aureus was eval-
uated at 37 °C in the glass chamber described above. Prior to the anti-
bacterial testing, the bacteria strains were cultured in the Luria-Bertani
(LB) culture medium overnight on a shaker (37 °C, 200 rpm), and then
diluted to the concentration of approximately 107 CFU/ml.
The as-prepared culture medium (2 ml) was added to 200 ml fresh
LB broth medium. After shaking softly, a volume of 15 ml homogeneous
broth was injected into each of the nine uncovered sterile petri dishes
(diameter 90 mm), respectively, and then the dishes with broth about
2.4 mm in thickness were placed in the corresponding chamber at one

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L. Li et al. Food Chemistry 269 (2018) 142–149

Fig. 4. Appearance changes of strawberries stored at 21 °C: (A) Control, (B) Exposed to GA fiber mats, and (C) Exposed to GA-BHA fiber mats.

end. Subsequently, GA-BHA fiber mats (6 × 6 cm) with a weight of
75 mg were laid aside the other end of glass chambers described above
(fiber mats without additional BHA as a control). The distance between
each dish and the fiber mat was about 40 mm. All the well-sealed
chambers were incubated on the shaker platform (37 °C, 60 rpm). At
each 4 h interval, the bacteria concentrations of the mediums were
determined and depicted an optical density at 600 mm (OD600) by using
the spectrophotometer. The inhibition efficiency (IE) of the fiber mats
against S. aureus was calculated as follows:

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L. Li et al.
ODC600−OD600
S
Inhibition efficiency (%) = × 100
C
OD600 (3)
where ODc600 and ODs600 represent the OD600 values of control and
culture medium at time t, respectively.
2.3.8. Application to strawberries
To evaluate the application of the GA-BHA fiber mats in active food
packaging, a representative fruits of strawberry was selected in our
experiment. Eighteen groups of strawberries (3 treatments × 6 re-
plicates, 4 fruits in each replicate) were put in the glass chambers de-
scribed above (initial relative humidity at 65%). Here, the three treat-
ments were as follows: GA fiber mats (6 × 6 cm, 75 mg), GA-BHA30
fiber mats (6 × 6 cm, 75 mg) and control (only strawberries). Similarly,
fiber mats were at one end of the well-sealed sterile chamber, straw-
berries at the other end. After then, these groups were stored at
21 ± 0.2 °C in the biochemical incubator and the appearance changes
of strawberries were observed at each 2-day interval.
For further investigating the inhibited species of microorganisms,
the strains on rotten strawberries were qualitatively screened by em-
ploying plate assay technique. Here, the strains were randomly col-
lected from the rotten strawberries and incubated in potato dextrose
broths (100 ml) for 36 h on the shaker platform (28 °C, 200 rpm).
Subsequently, serial 107-dilution cultures were evenly coated on the
potato dextrose agar medium to incubate at 28 °C for 36 h in the bio-
chemical incubator. Finally, the pure colonies were harvested and
viewed microscopically.
Schematic illustration of BHA volatile release and application to
strawberries were shown in Fig. 1.
2.4. Statistical analysis
Each experiment was carried out in triplicate. The results were re-
ported as means ± standard deviation (SD) and significance was con-
sidered at p < 0.05.
3. Results and discussion
3.1. Morphology and structure investigation
The fiber morphology and size are associated with BHA contents
(Fig. S3). As indicated from Table S1, the diameters of fibers increased
from 655 ± 6.0 (GA fibers) to 848 ± 5.9 nm (GA-BHA40 fibers).
Noteworthily, GA-BHA40 fibers were uneven and unsmooth on surface.
Therefore, a BHA content of 30% was considered as the optimum
content for the fabrication of uniform and continuous GA-BHA fibers.
Generally, the fiber morphology and size were directly associated with
electrospinning solution viscosity and conductivity. Table S1 showed
that the addition of BHA increased the solution viscosity while the
electrical conductivity change was not obvious. The increase in visc-
osity might be attributed to the intermolecular hydrogen bonds which
formed between GA and BHA. The splitting of droplets was more dif-
ficult at a higher viscosity during electrospinning, as a result, the fibers
became larger (Chiou et al., 2013).
Fig. 2A shows the FTIR spectra of BHA powders, GA fiber mats and
representative fiber mats (GA-BHA30), respectively. In GA mats spec-
trum, the characteristic absorptions could be identified as follows: NeH
and eOH stretching (3309 cm−1), CeH stretching (3080 cm−1), and
CeH stretching vibrations of aliphatic groups (2969, 2938 and
2883 cm−1); C]O stretching in amide I (1631 cm−1), coupling of NeH
bending and CeN stretching in amide II (1529 cm−1), and NeH
bending in amide III (1234 cm−1) (Vogt et al., 2013). In BHA powders
spectrum, the characteristic absorptions appeared at: –OH stretching
(3384 cm−1) and phenyl ring vibration (1504 cm−1); CeOeC sym-
metric stretching or symmetric bending of phenyl (1033 cm−1) and
vibration out-of-plane ring (815, 773 and 682 cm−1). Comparisons with
147
Food Chemistry 269 (2018) 142–149
the spectra of GA and BHA, all the characteristic absorption peaks of GA
and BHA appeared in the spectrum of GA-BHA fiber mats. Meanwhile,
the OeH stretching of BHA showed a lower-shift from 3384 to
3297 cm−1, and the NeH bending of GA presented a lower-shift from
1234 to 1229 cm−1. This information indicated the formation of in-
termolecular hydrogen bonds between GA and BHA in GA-BHA fibers as
depicted in Fig. 2B.
Fig. 2C illustrates CD spectra of GA-BHA solutions. Each curve
presented a weak positive peak at about 222 nm for the triple-helical
structure of GA and an obvious negative peak at about 199 nm for the
random coil conformation (Liu et al., 2017). Furthermore, a slight in-
crease of the positive peak and a relatively larger decrease of the ne-
gative peak suggested the encapsulation of BHA favored the con-
formational transition of coil to helix of GA. According to Fig. 2C, the
secondary structure proportions of GA were listed in Table 1. The
contents of α-helix and β-sheet structures increased with the increase of
BHA contents, while the random coil and β-turns structures decreased.
It could be concluded the hydrogen bonds formed between GA and BHA
would be in favor of the transition of random coil and β-turns to α-helix
and β-sheet.
Fig. 2D shows the influence of BHA on thermal property of the fiber
mats. Visibly, each fiber had a similar thermal behavior and presented a
characteristic endothermic peak termed denaturation temperature (TD)
which reflected the structure transition of helix to coil (Deng et al.,
2018). Additionally, the values of TD increased from 87.5 °C (GA) to
92.6 °C (GA-BHA40), indicating the encapsulation of BHA could slightly
enhance the thermal stability of GA matrix. Under the same condition,
the secondary structure (α-helix and β-sheet) change of GA in liquid
state could reflect that in solid state. From the CD results in liquid state,
we could speculate that higher contents of α-helix and β-sheet struc-
tures maybe enhance the protein chain stability.
3.2. Encapsulation efficiency of fibers
Encapsulation efficiency (EE) of BHA in GA fibers was obtained
according to eq (1) and summarized in Table S1. The EE data above
90% indicated that a high-efficiency encapsulation of BHA in GA fibers
could be achieved facilely via electrospinning. Similarly, Deng and co-
workers reported curcumin loaded GA electrospun fibers with an EE
value of almost 100% (Deng et al., 2017). Comparison with the en-
capsulation of curcumin, the slightly lower EE value of BHA was as-
sociated with its higher volatility. Maybe the diffusion of BHA in fiber
matrix was mainly driven by BHA content, the EE values of BHA in
fibers showed a slight decrease with increasing BHA contents.
3.3. BHA volatile release from fiber mats
The volatile release behavior of BHA from GA-BHA fiber mats was
investigated by using CV (Fig. 3A, representative fiber mats GA-BHA30,
the other two in Fig. S4). According to the CV curves, the released
amounts of BHA to the space (Fig. 3B) were determined by using
equation (Fig. S2B, inset). Each curve presented a similar variation
trend: an initial burst release within first 6 h and then gradually re-
leasing before reaching one plateau. This trend indicated the volatile
release of BHA from GA fiber mats was well-controlled. The burst re-
lease was majorly due to the volatile release of BHA absorbed or loosely
bound on or near the fiber surface (Wang et al., 2016). After then, BHA
molecules entrapped in the inner core of fiber matrices took a longer
time to release and resulted in a gradual volatile release. As the release
space is the same, the released amount of BHA from each fiber mats was
almost equal at equilibrium. Besides, a higher BHA content favored the
volatile release before the equilibrium because of the diffusion driving
force derived from BHA content.
For further investigating the volatile release mechanism of BHA
from fiber mats, the five empirical models (Ritger and Peppas, Zero-
order, First-order, Higuchi, and Hixson-Crowell) as shown in Figs.
L. Li et al.
S5(B–F) were employed on basis of Fig. S5A (Mt/M∞ vs t). The release
exponent (n) of Ritger and Peppas model and the correlation coeffi-
cients (R2) of each model were summarized in Table 2. The n values
below 0.45 (GA-BHA20: 0.43, GA-BHA30: 0.32, GA-BHA40: 0.23) im-
plied the major release was Fickian diffusion (Costa & Sousa Lobo,
2001; Ritger & Peppas, 1987a). The diffusion may be owing to BHA
volatile release partly through GA fiber matrix and partially through the
channels in the fiber structures; moreover, the n values decreased with
the increase of BHA contents, suggesting that a lower content of BHA
favored Fickian diffusion. As indicated from the values of R2, First-order
model is the best fit for the release data (R2 from 0.9976 to 0.9954),
indicating that the BHA transport in fiber matrix maybe a rate-de-
termining step; meanwhile, the release behavior was also agreed well
with Hixson-Crowell model (R2 from 0.9715 to 0.8891). Thereby, the
volatile release of BHA from GA fiber mats may be more inclined to a
diffusion mechanism.
3.4. Antioxidant activity of BHA in fibers
The presence of anodic peak between 0.59 and 0.63 V in each cyclic
voltammogram (Fig. 3C) was attributed to the oxidation of eOH group
on benzene ring (Wang et al., 2016), which indicated that BHA could
retain its antioxidant activity after being encapsulated in GA fibers,
suggesting that electrospinning is an effective method for the en-
capsulation of antioxidants. Additionally, the high linearity
(R2 = 0.9996) between anodic peak current (ip) and square root of scan
rate (v1/2) (Fig. 3D) implied that the electrooxidation process of BHA on
the glassy carbon electrode surface was diffusion-controlled, which
could be preceded by chemical reaction (Prabakar & Narayanan, 2006).
3.5. Antibacterial activity of GA-BHA fiber mats
Fig. 3E shows the growth curves of S. aureus exposed to the BHA
volatile from GA-BHA mats in chamber, Fig. 3F for the corresponding
inhibition efficiencies. As expected, three representative dynamic
phases composed of lag, exponential, and stabilization phases were
presented at the growth curve of S. aureus for the control. At the initial
lag phase, bacteria number increased slowly because they needed to
adapt themselves to the new external environmental. After exposure to
the BHA volatile, some of the bacteria were killed through touching.
Hence, the curves presented a slight decline with increasing BHA con-
tents. At exponential phase, the bacteria reproduced as an exponential
function of time owing to adequate nutrients and less metabolic pro-
ducts. The number of bacteria exposed to the BHA volatile significantly
decreased with the increase of BHA contents as compared to the con-
trol, indicating a good antibacterial activity against S. aureus. As the
bacteria consumed more nutrients as time went on, the growth curve of
the control increased slowly before reaching a plateau at stabilization
phase. Different from the control, the growth curves of bacteria exposed
to the BHA volatile were significantly declined at this phase, especially
for the mats with a higher BHA contents. Each inhibition efficiency
curve (Fig. 3F) shows a similar trend at the three phases. The relatively
higher inhibition efficiency at lag phase was due to the lower level of
bacteria number. As incubation time prolonged, the bacterial number
increased rapidly at exponential phase, while the released amount of
BHA from GA fibers was not enough to inhibit the bacteria multiply.
Therefore, a decline of IE appeared at the curves. After then, the growth
rate of bacteria decreased and the amount of bacteria was inclined to be
stable at stabilization phase, at this moment, the amount of the BHA
volatile in the chamber reached a higher value. Correspondingly, the
inhibition efficiency of BHA increased once more.
3.6. Application to strawberry preservation
The representative GA-BHA30 mats were applied to strawberry
preservation to evaluate the potential in active food packaging (Fig. 4).
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Food Chemistry 269 (2018) 142–149
On the 4th day, the control decayed similar to the work reported by
Wen et al. (2016) and the experimental group with GA mats also de-
cayed indicating that GA has no inhibitory effect on the microbes.
While the experimental group with GA-BHA30 mats showed no sign of
decay until to the 10th day. Hence, the GA-BHA fiber mats could ef-
fectively prolong the shelf-life of strawberries.
We noticed that colonies grew on the surface of the rotten straw-
berries on the 8th day for the control and the experimental group with
GA mats. To determine the species of microorganisms, the strains were
qualitatively screened and the results were illustrated in Fig. S6. Four
mould genera (Rhizopus sp., Mucor sp., Aspergillus sp. and Penicillium
sp.) were examined and isolated from the rotten strawberries, more-
over, Aspergillus niger (A. niger) and Aspergillus flavus (A. flavus) were
harvested from the Aspergillus sp. The results showed that GA-BHA fiber
mats also had broad-spectrum antifungal activity.
4. Conclusions
The functionalized GA-BHA fiber mats were successfully fabricated
via electrospinning technique. The fiber morphology and size are de-
pendent on BHA contents, and the interaction between GA and BHA
molecules was hydrogen bonds. These bonds favored the structure
transition of GA from random coil and β-turns to α-helix and β-sheet,
which enhanced the stability of protein chains. BHA was efficiently
encapsulated in GA fibers and the antioxidant activity was well-pro-
tected. The volatile release of BHA from GA-BHA mats was well-con-
trolled, and the predominant release mechanism was Fickian diffusion.
The application to strawberry showed that the GA-BHA mats had broad-
spectrum antifungal activity besides the better antibacterial activity.
Conflict of interest
The authors declare that no conflict of interest exists.
Acknowledgment
The research was supported by National Natural Science Foundation
of China, China (31371859).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in the
online version, at https://doi.org/10.1016/j.foodchem.2018.06.150.
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