Food Chemistry 231 (2017) 7077

Contents lists available at ScienceDirect

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

Effects of surfactants on the formation of gelatin nanofibres for

controlled release of curcumin
Lingli Deng, Xuefan Kang, Yuyu Liu, Fengqin Feng, Hui Zhang
College of Biosystems Engineering and Food Science, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang R&D Center for Food Technology
and Equipment, Zhejiang University, Hangzhou 310058, China

a r t i c l e i n f o a b s t r a c t

Article history: This work studied the effects of non-ionic Tween 80, anionic sodium dodecyl sulfonate (SDS) and cationic
Received 14 November 2016 cetyltrimethyl ammonium bromide (CTAB) surfactants on the morphology of electrospun gelatin nanofi-
Received in revised form 20 February 2017 bres, and on the release behaviour, antioxidant activity and antimicrobial activity of encapsulated cur-
Accepted 6 March 2017
cumin. Scanning electron micrographs showed that addition of SDS significantly increased the
Available online 8 March 2017
nanofibre diameter. Fourier transform infrared and differential scanning calorimetry analysis indicated
that gelatin and SDS intimately interacted via electrostatic and hydrophobic interactions. However, these
Keywords:
interactions inhibited the release of curcumin from the nanofibres with SDS, while CTAB and Tween 80
Electrospinning
Gelatin
both facilitated the release. SDS and Tween 80 showed protective effects on curcumin from the attack of
Surfactants 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radicals, and the increased release of curcumin from
Curcumin nanofibres with CTAB or Tween 80 resulted in a higher reducing power. The antimicrobial activity results
suggested that the curcumin encapsulated gelatin nanofibres with CTAB exhibited effective inhibition
against Staphylococcus aureus.
2017 Elsevier Ltd. All rights reserved.

1. Introduction
Electrospinning is a novel fabrication technique to produce
nanofibres, which has been applied in bioactive delivery, active
packaging, enzyme immobilization, and filtration (Aytac, Kusku,
Durgun, & Uyar, 2016; Sti et al., 2016; Wen et al., 2016). Based
on the safety concerns in food industry, it is necessary to produce
nanofibres that are solely or mainly composed of food biopoly-
mers. During the past few decades, natural biopolymers such as
collagen, gelatin, zein, chitin, and alginate have been successfully
electrospun solely or mixed with synthetic polymers (Geng,
Kwon, & Jang, 2005; Li, He, Zheng, & Han, 2006; Nie et al., 2008;
Selling et al., 2007). Gelatin is one of the most commonly used
GRAS (Generally Recognized as Safe) biopolymers because of its
biocompatibility and easy availability. Although gelatin is soluble
in water, it cannot be electrospun from aqueous solutions at room
temperature due to extensive hydrogen bonding resulting in gel
formation, low volatility of water, and excessively high surface ten-
sions (Moon & Farris, 2009). Therefore, toxic and aggressive sol-
vents such as 1,1,1,3,3,3-hexafluoro-2-propanol, 2,2,2-
trifluoroethanol, and formic acid have been used to electrospin
Corresponding author.
E-mail address: hubert0513@zju.edu.cn (H. Zhang).
gelatin (Ki et al., 2005; Song, Kim, & Kim, 2008). Rather than using
organic solvents that are unsuitable for food applications, food-
approved solvents should be applied (Laha, Yadav, Majumdar, &
Sharma, 2016; Okutan, Terzi, & Altay, 2014; Sen & Culha, 2016).
There have been a few reports on fabricating gelatin nanofibres
with acetic acid as a food-approved solvent for the purpose of
encapsulating bioactives. Hani, Torkamani, Azarian, Mahmood,
and Ngalim (2017) encapsulated Moringa oleifera bioactive extract
in gelatin nanofibres spun from 30% acetic acid solution to improve
its stability and maintain the antioxidant activity. Laha et al. (2016)
fabricated the crosslinked gelatin nanofibres from 20% acetic acid
solution, and the controlled release experiments suggested good
compatibility of hydrophobic molecules in the fibres.
Curcumin, a typical hydrophobic polyphenol, is characterized
by a variety of properties such as antioxidant, antimicrobial activ-
ity, and low toxicity (Anand, Kunnumakkara, Newman, &
Aggarwal, 2007). Numerous studies have shown that encapsulating
curcumin within delivery systems, such as liposome and nanopar-
ticles, could significantly improve the bioavailability of curcumin
in vitro and vivo (Kadota et al., 2016; Shaikh, Ankola, Beniwal,
Singh, & Kumar, 2009). Recently, studies were conducted to encap-
sulate curcumin within electrospun nanofibres by synthetic poly-
mers (polycaprolactone-polyethylene, polylactide) (Yakub et al.,
2016), biopolymers (zein) (Bui, Chung, & Park, 2014), and mixtures
of them (poly caprolactone/gelatin) (Fallah, Bahrami, & Ranjbar-

http://dx.doi.org/10.1016/j.foodchem.2017.03.027
0308-8146/ 2017 Elsevier Ltd. All rights reserved.
 L. Deng et al. / Food Chemistry 231 (2017) 7077
Mohammadi, 2016). On the other hand, surfactants dispersed in
aqueous solutions, above the critical micelle concentration, can
spontaneously form micelles, which have been extensively utilized
for solubilization of hydrophobic bioactives like curcumin. Dvores,
Marom, and Magdassi (2012) presented a method to incorporate
hydrophobic propylparaben within poly(ethylene oxide) (PEO)
nanofibres by encapsulating propylparaben in anionic sodium
dodecyl sulfonate (SDS) micelles. Polyoxyethylene sorbitan mono-
oleate (Tween 80) and sorbitan monooleate (Span 80) were applied
to encapsulate lipophilic compounds to form nanoemulsions
through electrospinning, which might be used as a fast dissolving
delivery system (Gordon, Marom, & Magdassi, 2015).
In this study, typical non-ionic (Tween 80), cationic (CTAB), and
anionic (SDS) surfactants were added to gelatin solutions (40%
acetic acid aqueous solution), which were subjected to electrospin-
ning. The influence of surfactant type and concentration on solu-
tion properties, and the morphologies of the fabricated
nanofibres were evaluated. Differential scanning calorimetry
(DSC) and Fourier transform infrared (FTIR) were used to study
the interaction between gelatin and surfactants. The release beha-
viours of curcumin from nanofibres were studied in PBS or PBS/
ethanol mixture. The antioxidant activities of the curcumin loaded
nanofibres were analyzed by DPPH radical scavenging activity and
FRAP (ferric reducing antioxidant power) assays. The antimicrobial
effects against Escherichia coli and Staphylococcus aureus were eval-
uated by disc diffusion method.
2. Materials and methods
2.1. Chemicals
Polyoxyethylene sorbitan monooleate (Tween 80), cetyltri-
methyl ammonium bromide (CTAB), sodium dodecyl sulfate
(SDS), 2,4,6-tri(2-pyridinyl)-1,3,5-triazine (TPTZ), and 2,2-diphe
nyl-1-picryl-hydrazyl-hydrate (DPPH) were obtained commer-
cially from Sigma Aldrich (St. Louis, MO, USA). Gelatin (type B,
alkali processed, isoelectric point pH = 4.7) was purchased from
Aladdin Inc. (Shanghai, China). The other reagents were purchased
from Sinopharm Chemical Reagent Co., Ltd., China. All reagents
were used without further purification and the water used was
double-distilled.
2.2. Microorganisms
Microbial cultures of Staphylococcus aureus (CMCC 26003) and
Escherichia coli (ATCC 25922) were obtained from Qingdao Hope
Bio-Technology Co., Ltd, and preserved in the Department of Food
Science and Nutrition, Zhejiang University. The strains were cul-
tured in NA (Nutrient Agar, Hangzhou Microbiological Agents Co.,
Ltd, China) broth at pH 7.0 and transferred every 2024 h with
incubation at 37 C.
2.3. Nanofibre fabrication
Solutions for electrospinning were prepared by dissolving 25%
(w/v) gelatin in 40% acetic aqueous solution (G25A40). All the sur-
factants were added at concentrations of 1%, 2%, 3%, 4%, 5% (wt%)
with respect to the weight of gelatin. Curcumin was added at a
concentration of 1% with respect to the weight of gelatin. All the
solutions were stirred overnight to ensure complete dissolution.
Prior to electrospinning, the gelatin solutions were character-
ized for their shear viscosity, conductivity and surface tension.
The shear viscosity at 100 s
1 was measured by a stress-
controlled rheometer (MCR 302, Anton Paar, Austria) using a
cone-plate geometry with 50 mm diameter and 1 angle. Conduc-
71
tivity measurements were performed at 25 C using a DDS-307
conductivity meter (Shanghai Precision & Scientific Instrument
Co., Ltd, China). The surface tension was measured at 25 C using
the pendant drop method through a surface tension meter (OCA
15, Dataphysics, Germany). All measurements were conducted in
triplicate.
The syringe loaded with the gelatin solution was fitted with a
20 G steel needle and the solution was pumped at a flow rate of
0.5 ml/h using a syringe pump (LSP02-1B, Baoding Longer Preci-
sion Pump Co., Ltd., China). A voltage of 15 kV (Gamma High Volt-
age, USA) was applied and the tip-collector distance was kept at
10 cm. The produced nanofibres were collected on grounded alu-
minum foil. The electrospinning conditions were kept at 25 C with
a humidity of approximately 50% throughout the experiments.
2.4. Morphology of nanofibres
Morphologies of the nanofibres were observed using a field
emission scanning electron microscope (SU8010, Hitachi, Japan).
The average fibre diameters and their diameter distributions were
determined by measuring 40 fibres for each image from 5 ran-
domly selected SEM images using Nano Measure software.
2.5. Thermal analysis
Differential scanning calorimetry (Mettler-Toledo, Switzerland)
was performed under a nitrogen atmosphere at a flow rate of
50 ml/min. Samples were sealed in 40 ll aluminum pans and
heated from 30 to 250 C, with a heating rate of 10 C/min.
2.6. FTIR analysis
Fourier transform infrared (FTIR) absorption spectra of the
nanofibres was recorded using a Nicolet 170-SX instrument
(Thermo Nicolet Ltd., USA) at a resolution of 2 cm
1 with 32 scans
over the wavenumber range of 4004000 cm
1. Deconvolution of
infrared spectra was performed using Peakfit, Version 4.12 (Sea-
Solve Software Inc., United States). Band assignments in the amide
III region (12001350 cm
1) were detected according to the previ-
ous methods (Cai & Singh, 1999; Fu, Deoliveira, Trumble, Sarkar, &
Singh, 1994). All FTIR experiments were performed in triplicate.
2.7. Encapsulation efficiency
The encapsulation efficiency of curcumin in gelatin nanofibres
was determined as previously reported by Wang et al. (2017).
5 mg of nanofibres were first washed with 1 ml of water to remove
the surface curcumin, and then dissolved in 1 ml 50% ethanol aque-
ous solution for 24 h. The concentration of curcumin was measured
by determining the absorbance at 435 nm with a Tecan Infinite
M200 Pro instrument (Mannedorf, Switzerland). The encapsulation
efficieny (EE) of curcumin in nanofibres was calculated as follows:
Actual curcumin amount
EE %  100 1
Theoretical curcumin amount
Actual curcumin amounts were determined from regression
equation (y = 31.688x
1.1026, R2 = 0.999).
2.8. In vitro release study
In vitro released experiment was conducted according to the
method reported by Llorens, Ibaez, Del Valle, and Puiggal
(2015). The controlled release of curcumin from nanofibres was
investigated in PBS (0.01 M, pH 7.4) and PBS/ethanol (7:3 v/v) mix-
ture as a more hydrophobic medium. The curcumin loaded fibres
(5 mg) were placed into 5 ml of PBS or PBS/ethanol mixture at
72 L. Deng et al. / Food Chemistry 231 (2017) 7077

37 C under shaking at 100 rpm. Samples were withdrawn from
the release medium at predetermined time intervals. The release
of curcumin was analyzed by determining the absorbance at
435 nm with a Tecan Infinite M200 Pro instrument (Mannedorf,
Switzerland). The volume was kept constant by adding fresh med-
ium. All release tests were carried out in triplicate.
2.9. Antioxidant activity
The radical scavenging activity of curcumin loaded fibre mats
was measured according to the method of Aytac and Uyar
(2016), with slight modifications. The curcumin loaded fibres
(5 mg) were weighed into cuvettes, in which 3 ml of freshly pre-
pared DPPH (10 -4 M) methanol solution was quickly added. The
absorbance change at 517 nm on a UVVis spectrometer (Shanghai
Spectrum Instruments Co. Ltd., China) within 30 min were
recorded by a digital camera, and then analyzed by Corel Video
Studio to generate the dynamic absorbance curve. The blank was
prepared as above by replacing the test sample with equivalent
methanol. The radical scavenging activity (RSA%) was calculated
as follows:
AbB
Abs
RSA %  100 2 mation of gelatin/micelle aggregates. The electrostatic and
AbB
where AbB and AbS are the absorbance values of the blank and sam-
ple, respectively. The measurements were carried out in triplicate.
FRAP (Ferric reducing antioxidant power) of the curcumin
encapsulated nanofibres was measured as described by Benzie
and Strain (1996). The FRAP reagents was prepared freshly by mix-
ing 10 mM TPTZ, 20 mM FeCl3 and 300 mM acetate buffer (pH 3.6)
in a ratio of 1:1:10 (v/v/v). The curcumin loaded fibres (5 mg) were
weighed into cuvettes, into which 1 ml of FRAP reagents was
quickly added. The mixtures were shaken, incubated at 37 C in
the dark for 30 min and then the absorbance at 593 nm was
recorded using a Tecan Infinite M200 Pro instrument (Mannedorf,
Switzerland). The measurement was compared to a standard curve
for FeSO4 solutions and expressed as the lmol of Fe2+ produced per
gram of the fibre. The nanofibres without curcumin were used as
blank. The measurements were carried out in triplicate.
3. Results and discussion
3.1. Solution properties
Addition of a surfactant to the polymer solution may modulate
the polymer/solvent and polymer/surfactant interactions via elec-
trostatic, hydrophobic interactions, and hydrogen bonding, so as
to alter the surface tension, viscosity, and conductivity of polymer
solutions (Kriegel, Kit, McClements, & Weiss, 2009b). The conduc-
tivity, viscosity, and surface tension of gelatin solutions with SDS,
CTAB, and Tween 80 added at various concentrations are presented
in Table 1. Results showed that the solution conductivity was sig-
nificantly increased by the addition of ionic surfactants rather than
non-ionic surfactant Tween 80. The increase of viscosity is depen-
dent on the surfactant concentration and type. The addition of 5%
CTAB, Tween 80, and SDS increased the viscosity of gelatin solu-
tions from 161.7 mPas to 185.7 mPas, 189.0 mPas, and
204.0 mPas, respectively. In terms of surface tension, the addition
of 1% SDS, CTAB, and Tween 80 caused a sudden decrease from
37.86 mN/m to 31.08 mN/m, 19.24 mN/m, 19.76 mN/m, respec-
tively, but the surfactant concentration had no effect. This kind
of gelatin-surfactant interaction at air/water interface has been
studied by Wu, Xu, Feng, and Li (2007). It can be assumed that
gelatin molecules absorb at air/solution interface, addition of sur-
factants above the critical micelle concentration leads to the for-
hydrophobic interactions between gelatin and surfactants cause
all gelatin molecules at the interface to move into the bulk. Ulti-
mately, the interface is occupied by surfactant molecules, resulting
in a sudden decrease in surface tension of the gelatin solution.
Gelatin in acetic acid carries a strong positive charge, which is
expected to interact electrostatically with ionic surfactants. Since
surfactants were added above the critical micelle concentrations,
micelles rather than monomers may interact with the polymer
chains (Griffiths, Stilbs, Howe, & Whitesides, 1996). The binding
between the anionic SDS micelles and the positively charged gela-
tin molecules could decrease the repulsive interactions between
individual gelatin molecules, thus facilitating increased entangle-
ments between gelatin chains, which might be an explanation for
the increased viscosity upon addition of SDS (Kriegel et al.,
2009b). On the other hand, CTAB is a positively charged surfactant
whose head groups should be electrostatically repelled from the
cationic groups of gelatin. Nevertheless, CTAB may still bind to

Table 1
Conductivity, viscosity, and surface tension of 25% gelatin in 40% acetic acid with SDS,
2.10. Antimicrobial activity CTAB, and Tween 80 added at concentrations of 1%, 2%, 3%, 4%, and 5% (relative to the
weight of gelatin).
The gram-positive S. aureus and the gram-negative E. coli bacte- Sample Conductivity Viscosity Surface tension
ria were used as model microbes to analyze the antimicrobial (mS/cm) (mPas) (mN/m)
activity of the curcumin loaded nanofibres by disc diffusion G25A40* 2.39 0.02a 161.7 1.2a 37.86 0.04a
method (Wen et al., 2016). The bacterial suspension (100 ll) of 1% SDS 2.46 0.01b 167.7 1.5b 31.08 0.75b
the inoculums (106 cfu/ml) was spread uniformly on the solidi- 2% SDS 2.54 0.01c 180.0 1.0c 30.93 0.07b
fied NA plate with a sterilized spreader. After that, nanofibre mats 3% SDS 2.63 0.01d 190.3 1.5d 31.17 0.22b
4% SDS 2.71 0.01e 205.0 2.0e 30.82 0.16b
cut into 10 mm disks (0.05 mm in thickness) were placed above 5% SDS 2.83 0.01f 204.0 2.0e 31.77 0.15b
the inoculated agar. The plates were then incubated at 37 C for 1% CTAB 2.51 0.01b 162.3 0.6a 19.24 0.11b
24 h to record the inhibition zones. 2% CTAB 2.57 0.01c 167.0 2.0b 19.30 0.05b
3% CTAB 2.66 0.01d 169.3 1.5c 19.15 0.05b
4% CTAB 2.75 0.01e 176.3 0.6d 19.21 0.06b
5% CTAB 2.82 0.01f 185.7 1.2e 19.29 0.04b
1% Tween 80 2.41 0.01a 175.0 1.0b 19.76 0.03b
2.11. Statistical analysis
2% Tween 80 2.39 0.01a 175.0 1.2b 20.06 0.08b
3% Tween 80 2.37 0.01a 176.0 1.0b 19.66 0.09b
All quantitative data were statistically analyzed to express as 4% Tween 80 2.39 0.01a 182.7 1.2c 19.89 0.04b
the mean standard deviation (SD). The statistical significance 5% Tween 80 2.39 0.01a 189.0 1.5c 20.07 0.05b
of mean values between multiple treatment groups was accessed Results are presented as mean values standard deviation of 3 replicates. Different
by one-way analysis of variance (ANOVA) with Tukeys test. P letters indicate significant difference (p < 0.05) between each parameter tested.
*
value < 0.05 was considered statistically significant. G25A40 is the abbreviation of 25% (w/v) gelatin in 40% acetic acid solution.
 L. Deng et al. / Food Chemistry 231 (2017) 7077
gelatin through hydrophobic interactions between its non-polar
tail and non-polar groups on gelatin molecules (Mitra,
Bhattacharya, & Moulik, 2009). Tween 80 as a nonionic surfactant
would be expected to show no electrostatic interaction and instead
may interact with gelatin solely via hydrophobic interaction. The
fact that the addition of Tween 80 or CTAB to the gelatin solution
caused little change in viscosity suggests that it did not strongly
affect the interactions between the gelatin molecules (Kriegel,
Kit, McClements, & Weiss, 2009a).
3.2. Morphology of the nanofibres
When surfactants were added to the gelatin solution, smooth
nanofibres were formed at all concentrations. Addition of CTAB
and Tween 80 did not significantly affect the diameter of gelatin
nanofibres (Figs. S1 and S2). However, addition of SDS yielded
nanofibres with increased diameters (Fig. 1). The increase in fibre
diameter was concentration dependent. When 5% SDS was added,
the average diameter was 368 nm, while the nanofibre without
surfactants had an average diameter of 205 nm. Surface tension,
viscosity, and conductivity are of key importance to the process
of electrospinning influencing the diameter distribution of nanofi-
bres. At lower surface tensions, there is decreased energy of the
system forcing a breakup of the polymer jets into spheres. The for-
mation of thinner nanofibres is also opposed by viscoelastic force.
On the other hand, higher conductivities may result in increased
charge densities being present at the fibre surface, which may
favour an increase in surface area and thus oppose the formation
of beads and promote thinner jets (Kriegel, Arrechi, Kit,
McClements, & Weiss, 2008). Although the increase in conductivity
and decrease in surface tension favoured the stretching of nanofi-
bres, the significant increase in viscosity retarded the stretching,
leading to the ultimate increase in fibre diameter. This is in agree-
ment with the reports of Hu, Prabhakaran, Ding, and Ramakrishna
(2015), who found that addition of SDS to polycaprolactone
increased the nanofibre diameter. Kriegel et al. (2009b) have also
found that higher SDS concentrations could cause an increase in
chitosan/PEO fibre diameter.
3.3. Characterization of nanofibres
The thermal behaviours of electrospun gelatin nanofibres and
those with 5% SDS, CTAB, and Tween 80 added were measured by
DSC (Fig. S3). It is evident that all fibres had a similar thermal beha-
viour such that they showed an endothermic melting transition (Tm)
within a relatively broad temperature range. The Tm of the gelatin
indicated the temperature caused a destruction of ordered or aggre-
gated structure (Tongnuanchan, Benjakul, & Prodpran, 2014). A
melting temperature of 91.20 C was observed for pure gelatin
nanofibres, while the addition of SDS caused an increase to
100.52 C. However, the addition of CTAB and Tween 80 did not
change the thermal properties of gelatin fibres. These results suggest
that SDS not only interacts intimately with gelatin in the network,
but also enhances the strong protein-protein interaction
(Tongnuanchan et al., 2014), which coincides with the solution
properties in the current study. Similarly, studies conducted by
Vasita, Mani, Agrawal, and Katti (2010) showed that the Tm of poly
(lactic-co-glycolic acid) (PLGA) increased after addition of the non-
ionic surfactant Pluronic F-108. Hu et al. (2015) found that addition
of SDS and benzyltriethylammonium chloride (TEBAC) led to a slight
increase in the Tm of poly (e-caprolactone) nanofibre mat.
The interaction between SDS and gelatin was studied by the
FTIR spectra (Fig. S4). In detail, the characteristic peaks were inter-
preted as follows: the stretching of NAH and hydrogen bonding at
32933305 cm
1 (amide A), the stretching of CAH at 3071
3076 cm
1, CAH stretching vibrations of aliphatic groups at
73
2874, 2931, 2959 cm
1, the stretching of C@O at 1641
1652 cm
1 (amide I), the bending and stretching of CAN at
15411544 cm
1 (amide II), NAH bend and C-N stretching combi-
nation band, NH3+ symmetric deformation at 1454 cm
1, stretch-
ing of ACAN at 14061412 cm
1, CH deformation of the methyl
group at 1336 cm
1, and the stretching of CAN and bending of
NAH at 12421247 cm
1 (amide III). It is obvious that the intensity
of amide A band increased after adding SDS, indicating the forma-
tion of hydrogen bonds between gelatin and SDS. Table 2 shows
the secondary structures of gelatin with various concentrations
of SDS added. With the addition of SDS, the ratio of b-sheet
increased while the ratio of turns decreased accordingly. The bind-
ing of SDS to the gelatin molecules leads to a complex that may
retain its overall charge, thereby affecting intermolecular interac-
tions and the gelatin conformation. In turn, the polymers may
assume a more open and expanded structure which could result
in nanofibres with an increased diameter (Kriegel et al., 2009b).
3.4. Release behaviour of curcumin
The encapsulation efficiency of curcumin in pure gelatin nanofi-
bre (95.0 4.3%) showed no significant difference with those fibres
with 5% SDS (94.6 4.7%), CTAB (95.8 3.4%), and Tween 80
(95.6 5.0%) added, respectively. This indicated that almost 100%
of curcumin was well protected after being loaded into gelatin
nanofibres. Fig. 2 presents the release behaviours of the curcumin
encapsulated nanofibres when exposed to a hydrophilic PBS and a
more hydrophobic PBS/ethanol mixture media. Release proceeded
at a similarly fast rate until a limit value is reached. The release rate
and ratio are highly dependent on the affinity between curcumin
and the polymer as well as affinity between curcumin and the
release medium (Llorens et al., 2015). The ratio of curcumin
released from the pure gelatin nanofibres to PBS within 4 h was
7.1% due to the solubility limit (Fig. 2a). CTAB and Tween 80 rather
than SDS increased significantly the release ratio of curcumin in
PBS. However, the total release ratio after 4 h was still limited in
PBS due to the hydrophobicity of curcumin. When a more
hydrophobic solvent (PBS/ethanol) was applied, curcumin was
released rapidly from nanofibres. The quick release of curcumin
near the fibre surface resulted in an initial burst release within
60 min for each nanofibre (Fig. 2b). The released ratio of curcumin
from the pure gelatin nanofibre was 49.8% at first 60 min, slightly
lower than those with 5% SDS and CTAB added. After that, CTAB
still promoted the curcumin release while the accumulative release
of other fibres reached a plateau.
It is clear that the addition of surfactants could improve the sol-
ubility of curcumin in a polar solvent by encapsulating curcumin
within the hydrophobic core of micelles, thus facilitating the
release of curcumin into PBS. However, there is no obvious
improvement observed for the gelatin nanofibre with SDS, which
could be attributed to the intimate interaction between SDS and
gelatin. Although curcumin has been encapsulated within SDS
micelles, the micelles were restricted in the network of gelatin
molecules, resulting in the slow release of curcumin. Stephansen,
Garca-Daz, Jessen, Chronakis, and Nielsen (2016) studied the
effect of surfactants on release behaviour of insulin from sarcoplas-
mic protein, and found that the anionic surfactant promoted the
release while the cationic surfactant showed contrary effects.
3.5. Antioxidant activity
The antioxidant activity of curcumin encapsulated gelatin
nanofibres was investigated by measuring DPPH radical scaveng-
ing activity and ferric reducing antioxidant power. Dynamic
changes of DPPH RSA after addition of nanofibres are shown in
Fig. 3a. Curcumin from the pure gelatin nanofibre showed the high-
74 L. Deng et al. / Food Chemistry 231 (2017) 7077

Fig. 1. Morphologies of (a) gelatin nanofibre, and those with SDS added at concentrations of (b) 1%, (c) 2%, (d) 3%, (e) 4% and (f) 5% (with respect to the weight of gelatin, w/w).

Table 2 the highest ferric reducing antioxidant power (96.6 lmol Fe2+/g),
Ratios of different secondary structures of nanofibres. compared to the pure gelatin fibre (38.0 lmol Fe2+/g) and those
b-sheet/% Turns/% a-helix/% with Tween 80 (71.4 lmol Fe2+/g) and SDS (30.3 lmol Fe2+/g).
G25A40 *
37.47 0.38a
24.31 0.23 a
38.21 0.60a It was known that the close proximity of radical and antioxidant
1% SDS 38.18 0.27ab 23.88 0.09ab 37.94 0.20ab is a crucial prerequisite for the radical reducing action of antioxi-
2% SDS 39.54 0.53c 23.07 0.60ab 37.39 0.09ac dants (Heins, McPhail, Sokolowski, Stckmann, & Schwarz, 2007).
3% SDS 40.01 0.47 cd 22.91 0.59b 37.08 0.15bcd Only after releasing from nanofibres, curcumin could react with
4% SDS 40.61 0.51 cd 22.66 0.40b 36.72 0.14 cd
the free radicals. CTAB favoured the release of curcumin from
5% SDS 40.92 0.23d 22.63 0.50b 36.44 0.31d
nanofibre to the hydrophobic solvent, leading to a higher radical
*
G25A40 is the abbreviation of 25% (w/v) gelatin in 40% acetic acid solution. scavenging activity and a ferric reducing antioxidant power than
other surfactants. Aytac et al. (2016) found that inclusion of a-
est reaction rate with RSA, compared to those with surfactants
tocopherol in b-cyclodextrin improved the DPPH RSA of PCL/a-
added. This could be attributed to the physical barrier effect of sur-
tocopherol nanofibre by accelerating the release of a-tocopherol
factant micelles that protect the curcumin from the attack of free
from the nanofibre.
radicals (Chat, Najar, Mir, Rather, & Dar, 2011). Within 30 min
the RSA of gelatin fibre and those with Tween 80, CTAB, and SDS
were calculated as 44.4%, 36.3%, 44.8%, and 34.8%, respectively 3.6. Antimicrobial activity
(Fig. 3b). Nanofibres with Tween 80 and SDS showed significant
lower RSA than the pure gelatin fibre, while the fibre with CTAB The antimicrobial activities of the curcumin encapsulated
did not differ. As shown in Fig. 3b, the nanofibre with CTAB had nanofibres against E. coli and S. aureus are shown in Table 3. No
 L. Deng et al. / Food Chemistry 231 (2017) 7077 75

Fig. 3. Dynamic change of DPPH radical scavenging activity after addition of (a)
curcumin encapsulated nanofibres and (b) DPPH radical scavenging activity and
ferric reducing antioxidant power of 1% curcumin encapsulated nanofibres with
various surfactants added at a concentration of 5% (with respect to the weight of
gelatin, w/w) within 30 min.

Fig. 2. Curcumin release from the gelatin nanofibre and those with various
surfactants added at a concentration of 5% (with respect to the weight of gelatin, w/
w) in vitro in (a) PBS (0.01 M, pH 7.4) and (b) PBS/ethanol (7:3).
Table 3
Inhibition zones of 1% curcumin encapsulated gelatin nanofibre and those with SDS,
CTAB, and Tween 80 added, compared to the blank control of nanofibres without
curcumin against E. coli and S. aureus.

Samples Inhibition zone diameter (mm)

inhibition zone was observed for all nanofibres with or without E. coli S. aureus.
curcumin against E. coli, except the fibre with CTAB. The inhibition Curcumin encapsulated G25A40 *
NA **
11.05 0.32a
zone against E. coli of the blank nanofibre with CTAB was SDS*** NA 14.96 0.96b
10.62 mm, while that of the curcumin encapsulated fibre was CTAB 11.99 0.93a 20.73 1.85d
11.99 mm. However, all nanofibres with curcumin showed effec- Tween 80 NA 11.30 0.21a
Blank control G25A40 NA NA
tive inhibition on S. aureus.
SDS NA 12.41 0.25c
The curcumin encapsulated nanofibres showed better antimi- CTAB 10.62 0.30a 16.66 0.28e
crobial activity against S. aureus than E. coli. The possible reason Tween 80 NA NA
is that the cell wall lipopolysaccharides of Gram-negative bacteria *
G25A40 is the abbreviation of 25% (w/v) gelatin in 40% acetic acid solution.
prevent active compounds from reaching the cytoplasmic mem- **
NA, not available.
brane (Basniwal, Buttar, Jain, & Jain, 2011). When the nanofibres ***
Surfactants were added at a concentration of 5% respect to the weight of gelatin.
are exposed to a humid environment, the surfactants and curcumin
diffuse out of the nanofibres, inhibiting bacterial growth in neigh-
bouring regions (Dvoracek, Sukhonosova, Benedik, & Grunlan, might be a synergy between curcumin and CTAB when inhibiting
2009). The fact that CTAB facilitates the release of curcumin is S. aureus, since CTAB has long been used as a safe antimicrobial
one possible reason for the highest antimicrobial activity. There cationic surfactant (Nakata, Tsuchido, & Matsumura, 2011).
76 L. Deng et al. / Food Chemistry 231 (2017) 7077
4. Conclusion
These results demonstrated that surfactants are able to modu-
late the morphology of gelatin nanofibres and affect the release,
antioxidant and antimicrobial activities of the encapsulated cur-
cumin. The addition of anionic SDS surfactant increased the fibre
diameter, as a more open and expanded structure was formed
for gelatin by interacting with SDS via electrostatic interaction
and hydrophobic interactions. However, the interactions between
SDS and gelatin did not favour the release of the encapsulated cur-
cumin, while CTAB and Tween 80 greatly improved the release of
curcumin into polar solvents. The fact that CTAB facilitates the
release of curcumin resulted in a higher radical scavenging activity
and a ferric reducing antioxidant power, as well as a stronger
antimicrobial activity. These results offer a new way to produce
gelatin nanofibres with food grade surfactants for the control
release of curcumin, which may find promising applications as a
nutraceutical carrier in food industry.
Acknowledgements
This work was financially supported by the National Natural
Science Foundation of China (Grant No. 31471622), and Fund of
Fuli Institute of Food Science Zhejiang University (Grant No.
KY201401).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.foodchem.2017.
03.027.
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