Food Research International 77 (2015) 467475

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Food Research International

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

Release of allyl isothiocyanate from mustard seed meal powder

entrapped in electrospun PLAPEO nonwovens
Ruyan Dai 1, Loong-Tak Lim
Department of Food Science, University of Guelph, ON N1G2W1, Canada

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

Article history: Defatted mustard seed meal powder (MSMP) is a natural source of allyl isothiocyanate (AITC) a potent antimi-
Received 29 May 2015 crobial volatile released from Brassicaceae plants when a glucosinolate (sinigrin) in the plant tissues is hydro-
Received in revised form 14 August 2015 lyzed in a myrosinase-catalyzed reaction. This study demonstrated the feasibility of entrapping MSMP particles
Accepted 18 August 2015
in nonwovens produced by electrospinning poly(lactic acid)poly(ethylene oxide) (PLAPEO) blends into ultra-
Available online 22 August 2015
ne bers. Although MSMP particles were larger than the electrospun PLAPEO bers, results from scanning elec-
Keywords:
tron microscopy showed that the mustard seed particles were physically entrapped and uniformly dispersed
Allyl isothiocyanate within the nonwovens. Moreover, the surface of the MSMP was fully covered by the polymer blend. By changing
Mustard seed meal powder the PLAPEO blend ratio, AITC release rate and quantity released could be manipulated readily. Specic interac-
Electrospun bers tions between PLA, PEO and MSMP were not detected with Fourier transform infrared spectroscopy. However,
Controlled release differential scanning calorimetry showed that components from MSMP interfered with the chain packing of
the crystalline phases of both polymers, lowering their melting temperatures. In view of the well-established an-
timicrobial properties of AITC, the RH-sensitive MSMP-containing nonwovens potentially can be useful in active
food packaging applications. Further studies involving specic foods will shed light on the extent by which the
active nonwovens affect product shelf-life.
2015 Elsevier Ltd. All rights reserved.

1. Introduction
Allyl isothiocyanate (AITC) is a naturally occurring compound found
in plants from the Brassicaceae family (e.g., mustard seed, horseradish,
and cabbage). Due to its broad-spectrum antimicrobial effects against
bacteria, molds, and yeast, AITC has been studied extensively for food
preservation (Delaquis & Mazza, 1995; Isshiki, Tokuoka, Mori, & Chiba,
1992; Kinae, Masuda, Shin, Furugori, & Shimoi, 2000; Mari, Leonib,
Iori, & Cembalia, 2002; Nadajarah et al., 2005; Shofran, Purrington,
Breidt, & Fleming, 1998). For this purpose, AITC has been added directly
to food products in the liquid phase, or apply as vapor in modied atmo-
sphere packaging (MAP) applications to extend the shelf-life of food
products, including fresh beef, cured pork, sliced raw tuna, cheese, egg
sandwich, noodles, and pasta (Jung et al., 2009; Parashar,
Ramakrishna, & Ramaprasad, 2011; Serrano et al., 2008; Shin, Harte,
Ryser, & Selke, 2010; Suppakul, Miltz, Sonneveld, & Bigger, 2003). Be-
cause AITC is volatile and possesses strong odor, it is important to opti-
mize its inhibitory concentration to prevent unwanted sensory issues,
while achieving adequate antimicrobial efcacy. To this end, AITC has
been encapsulated in various polymers to control its release, by
Corresponding author.
E-mail address: llim@uoguelph.ca (L.-T. Lim).
1
Author Dai is currently afliated with Weston Bakeries Ltd. & Ready Bake Foods Inc.,
Mississauga, ON, Canada.
manipulating the diffusivity and solubility of AITC in the polymeric ma-
trices (Ko, Jeon, & Park, 2012; Vega-Lugo & Lim, 2009). Alternatively, the
vapor pressure of AITC can be suppressed by blending it with a miscible
liquid, such as vegetable oil (Lim & Tung, 1997).
As an oilseed, mustard seed is pressed and ground for the extraction
of oil, resulting in mustard seed meal powder (MSMP) as a byproduct
(Marcone, Yada, Aroonkamonsri, & Kakadu, 1997). Since MSMP is rich
in the AITC precursor sinigrin, MSMP may be used as a natural source
of AITC in antimicrobial MAP applications to extend the shelf-life of
food. AITC is released from Brassicaceae plants when sinigrin in the
plant tissues is hydrolyzed in a myrosinase-catalyzed reaction
(Bjorkman & Lonnerdal, 1973). Because water is a required substrate
in this reaction, the release rate of AITC can be manipulated by control-
ling the accessibility of MSMP to water. Recently, we investigated the
feasibility of using MSMP as a natural source of AITC (Dai & Lim,
2014). In that study, we demonstrated that the natural matrix could
release 2 to 17 mg AITC/g of MSMP within 24 h, depending on the tem-
perature and relative humidity conditions used (85 to 100% RH, 5 to
35 C). Conceivably, when MSMP is exposed to the package headspace
of a product with elevated water activity, or when the product is
exposed to temperature abuse during transportation, the release of
AITC can be activated, thereby inhibiting the proliferation of spoilage
microorganisms.
In order to effectively deploy MSMP to deliver AITC in MAP applica-
tions, the particulate MSMP must be contained within a permeable

http://dx.doi.org/10.1016/j.foodres.2015.08.029
0963-9969/ 2015 Elsevier Ltd. All rights reserved.
468 R. Dai, L.-T. Lim / Food Research International 77 (2015) 467475
carrier (e.g., membrane, sachet, and capsule). By virtue of their sub-
micron diameter, electrospun bers possess a surface-to-volume ratio
considerably greater than other carriers such as those produced from
blown spun bers and cast lms. The large surface area of the
electrospun carrier is useful in applications where surface activation is
important. Moreover, by altering the composition of the bers, such as
blending with different polymers, their physicochemical properties (hy-
drophilicity, ber diameter, density, glass transition temperature) can
be manipulated readily to ne-tune the release rate of the encapsulated
species (Huang, Zhang, Kotakic, & Ramakrishna, 2003; Pham, Sharma, &
Mikos, 2006; Potinenia, Lynn, Langer, & Amiji, 2003; Schiffman &
Schauer, 2008; Sill & von Recum, 2008).
Poly(lactic acid) (PLA) is an aliphatic polyester derived from starch
and sugars, which exhibits material properties and processing charac-
teristics similar to many thermoplastics. Since PLA is biodegradable
and derived from renewable feedstock, it is being considered as an en-
vironmental friendly alternative to petroleum-based polymers (Lim,
Auras, & Rubino, 2008). By dissolving in a solvent, PLA can be
electrospun into ultrane bers for various applications, including en-
zyme immobilization, controlled release of drugs, and tissue scaffolds
(Vaz, van Tuijl, Bouten, & Baaijens, 2005; Yang, Murugan, Wang, &
Ramakrishna, 2005; Zhou & Lim, 2009; Zong et al., 2005). To optimize
the electrospun PLA bers for specic end-use applications, PLA is
often blended with other compatible polymers. For example, by blend-
ing PLA with poly(ethylene oxide) (PEO), composite bers that are
more hydrophilic than those made of pure PLA can be prepared
(Honarbakhsh & Pourdeyhimi, 2011).
The objectives of the present study are to: (1) develop PLAPEO non-
wovens, using an electrospinning methodology, to entrap MSMP parti-
cles; and (2) study the moisture-activated release of AITC from MSMP
particles entrapped in electrospun PLAPEO nonwovens.
2. Materials and methods
2.1. Materials
PLA (6201 D) was donated by NatureWorks LLC (Minnetonka, MN,
USA). PEO (300 kDa molecular weight), hexane, chloroform (HPLC
grade), and dimethyl formamide (DMF) were purchased from Sigma Al-
drich (Oakville, ON, Canada). MSMP was donated by Biofumes Technol-
ogies Inc. (Saskatoon, SK, Canada). AITC standard (N98% GC purity) was
purchased from Fluka Chemical Co. (Steinheim, Germany).
2.2. Methods
2.2.1. MSMP preparation
MSMP was defatted using the Soxhlet oil extraction method with
hexane, following by grinding into ne powder using a ball mill
(Model PM 100, Retsch GmbH, Haan, Germany). Ball milling was con-
ducted at 450 rpm for 1 h using eight stainless steel balls (diameter:
20 mm) in a 500 mL stainless steel grinding bowl. Sample preparation
was conducted at 24 2 C and 60 5% RH. Samples were freshly pre-
pared and stored in airtight glass jars for no more than two weeks be-
fore the encapsulation experiments. Particle size distribution and the
chemical composition of the resulting MSMP were analyzed as per Dai
and Lim (2014).
2.2.2. Preparation of polymer solution for electrospinning
Stock solutions of 9% (w/w) PLA and 3% (w/w) PEO were prepared
by dissolving PLA and PEO resins, respectively, in 9:1 (w/w)
chloroform:DMF mixture, and stirred for 12 h at 24 2 C using a mag-
netic stirrer. To study the effect of the PLA and PEO blend ratio on AITC
release behavior, the stock solutions were blended at 100:0, 75:25,
50:50, 25:75, and 0:100 ratios (w/w), coded as 100PLA0PEO,
75PLA25PEO, 50PLA50PEO, 25PLA75PEO, and 0PLA100PEO, respective-
ly. The defatted ground MSMP was then dispersed into the ve polymer
solutions at 9% (w/w) xed and mixed with the magnetic stirrer for 3 h
to form homogeneous suspensions for electrospinning.
2.2.3. Electrospinning
The suspensions prepared in Section 2.2.2 were electrospun using a
horizontal setup (Fig. 1). Suspensions were drawn into a 3 mL plastic sy-
ringe (KD Scientic, Holliston, USA) and pumped at 3 mL/h via a piston
assembled using a syringe pump (Orion M361, Thermo Scientic, Oak-
ville, ON, Canada). A 20-gauge blunt tip stainless steel needle, serving
as a spinneret, was attached to the syringe. The spinneret was connect-
ed to the positive electrode of a direct current power supply (DC model
ES50P-50W/DAM, Gamma High Voltage, Ormond Beach, FL, USA), oper-
ating at a constant voltage of 11 1 kV. A circular stainless steel plate
covered with aluminum foil was grounded to collect the electrospun -
bers. The distance between the tip of the spinneret and the collector was
xed at 15 cm. Electrospinning experiments were conducted in an envi-
ronmental chamber (Model MLR-350H, Sanyo Corp., Japan) maintained
at 25 0.5 C and 20 1% RH. At the end of the electrospinning process,
the nonwovens were transferred into a vacuum oven for 48 h at 40 C to
evaporate residual chloroform and DMF. Samples were stored in a des-
iccator (Fisher Scientic, Oakville, ON, Canada) before further
characterization.
2.3. Fiber morphology and size analysis
The microstructures of the nonwoven were evaluated using scan-
ning electron microscopy (SEM) (Model S-570, Hitachi High Technolo-
gies Corp., Tokyo, Japan). Electrospun samples were mounted on metal
stubs using double-adhesive carbon tape, and then coated with an ultra-
thin (20 nm) layer of gold and platinum using a sputter coater (Model
K550, Emitech, Ashford, Kent, UK). An accelerating voltage of 10 kV
was applied during the analysis. Samples were randomly selected
from three locations for analysis. Fiber diameters were analyzed with
image processing software (Image Pro-Plus 6.0, Media Cybernetics
Inc., Bethesda, MD, USA). For pristine ber samples, the diameters of
the bers were recorded, while for MSMP-containing samples, only
the brous portions were measured.
2.4. Attenuated total reection (ATR)Fourier transform infrared (FTIR)
analysis
FTIR spectra of pristine and MSMP-containing electrospun ber
samples were analyzed using a FTIR spectrometer (IRPrestige21,
Shimadzu Corp., Kyoto, Japan), equipped with an ATR cell (Pike Tech-
nologies, Madison, Wis., USA). Spectra were taken by averaging 40
scans for each sample at 4 cm1 resolution. Samples were mounted di-
rectly on the ATR sapphire crystal, compressed with a press, and
scanned between 700 and 3900 cm 1. Spectra were analyzed using
the IRsolution software (Shimadzu Corp., Kyoto, Japan). Three spots
were randomly selected from samples for analysis and each spot was
analyzed three times.
2.5. Thermal analysis
Thermal properties of the nonwovens were analyzed using a differ-
ential scanning calorimeter (DSC, Q2000, TA Inc. New Castle, DE, USA).
The DSC was calibrated using an indium standard (melting point:
156.6 C), and errors were found to be less than 2% in the heat ow cal-
ibration. To study the thermal characteristics of the nonwovens, sam-
ples were cut into small pieces, weighed accurately (3 to 5 mg) into
alodined-aluminum pans, and sealed. Samples were heated at a ramp
rate of 10 C/min in the range of 0 to 250 C. Thermal data were analyzed
using TA Universal Analysis Software (TA Inc. New Castle, DE, USA).
From the thermographs, the glass transition temperature (Tg) and melt-
ing temperature (Tm) were determined. Tg and Tm values were averages
of three determinations.
 R. Dai, L.-T. Lim / Food Research International 77 (2015) 467475
Fig. 1. Schematic diagram of the experimental setup for electrospinning of PLAPEO nonwovens.
2.6. Release of AITC through headspace analysis
The AITC release kinetic was studied using a gas chromatograph (GC
6890, Agilent Technologies Inc., Santa Clara, Calif., USA) equipped with a
ame ionization detector (FID), as described by Dai and Lim (2014).
MSMP-containing nonwovens (5 cm 8 cm) were cut and placed in
hermetically sealed 1 L glass jars containing a glass lter paper saturated
with 1 g of distilled water. A septum was attached to the lid of the jars,
allowing a sampling needle to pierce through the lid to extract the head-
space gas at predetermined time intervals. The glass jars were main-
tained at 20 0.5 C in an environmental chamber during the test.
Briey, the analysis involved lling the sampling loop of a gas sampling
valve with about 15 mL of headspace gas. When the valve was actuated,
the sampling loop was purged with nitrogen carrier gas, injecting the
headspace gas into a capillary column. Chromatograms were analyzed
using the Peak Simple software (39332 bit, SRI Instruments, CA,
USA). The calibration curve was prepared by injecting known amounts
of AITC standard into the glass jar and analyzed as above. To determine
the amount of AITC released at any given sampling point (Mc), the
amount of AITC removed during each previous sampling step was
taken account:
Mr Cr Vb
Ml Cr1  Ve Ml1 2
Mc Mr Ml
where Cr is the recorded AITC concentration calculated based on calibra-
tion constants at any given sampling point; Cr1 is the recorded concen-
tration of the previous sampling point; Vb is the volume of the glass jar;
Ve is the volume of headspace gas extracted; and Ml1 is the sum of
mass loss of all the previous sampling points before Ml.
2.7. Data analysis
Statistical analysis was conducted using the SPSS 20 software (IBM
SPSS Inc., Chicago, IL, USA). One-way ANOVA was carried out to detect
the differences of various factors and treatments at a 95% condence in-
terval with Tukey's post hoc test. All treatments were triplicated. AITC
469
release kinetics were modeled empirically using the modied Gompertz
equation (Lay, Lee, & Noike, 1999; Mu, Wang, & Yu, 2006):
  
k  exp1
Q t Q max exp exp t 1 4
Q max
where Qt is the amount of the AITC released at time t; Qmax is the max-
imum amount of AITC released; k is the rate constant; and is the lag
time. Non-linear regression analyses were conducted to determine
values for the three parameters using the Solver function in the
Microsoft Excel spreadsheet package (Microsoft Ofce 2011, WA,
USA). The GRG Nonlinear algorithm option was selected to minimize
the residual sum of squares by changing the three parameters, without
imposing any constraints.
3. Results and discussion
3.1. Microstructures of electrospun nonwovens
Figs. 2 and 3 are SEM micrographs taken at different levels of magni-
cation for pristine and MSMP-containing nonwovens. As shown,
electrospun bers for the pristine nonwovens had smooth surfaces. This
1 result is different from the electrospun PLAPEO bers observed by
Honarbakhsh and Pourdeyhimi (2011), which displayed porous
surface morphology, probably due to the higher vapor pressure of the
dichloromethane solvent used in their work, than that of the chloro-
formDMF solvent used in the present study. By contrast, MSMP-
3 containing bers displayed irregular entities along their length, which
can be attributed to the embedded MSMP particles that are larger than
the diameter of the electrospun bers. Approximately 75% of the MSMP
particles had radius in the range of 5 to 20 m, with a total of greater
than 95% of the particles having radius smaller than 30 m. Despite the
oversized MSMP particles, they were encapsulated by the polymer ma-
trix, as revealed by the absence of jagged surface morphologies seen in
unencapsulated MSMP powders (micrograph not shown). Also, as
shown in low magnication micrographs, there were no apparent particle
aggregations throughout the nonwoven. These results show that the PLA
PEO nonwovens are compatible matrices for entrapping MSMP particles.
In the absence of MSMP, the average diameters for the electrospun
bers ranged from 1.09 0.31 to 2.02 0.44 m, while for those dis-
persed with MSMP ranged from 0.77 0.37 to 1.50 0.46 m
470 R. Dai, L.-T. Lim / Food Research International 77 (2015) 467475

Fig. 2. Scanning electron micrographs of pristine electrospun ultrane bers: (a) 100PLA0PEO; (b) 75PLA25PEO; (c) 50PLA50PEO; (d) 25PLA75PEO; and (e) 0PLA100PEO. The histograms
show the distribution of ber (y-axis; fraction) in different diameter intervals (x-axis; m).

(Table 1). The smaller bers observed with the MSMP-containing bers
were probably caused by the increased momentum of the polymer jet as
it ejected from the spinneret tip, due to the extra mass carried by the
embedded MSMP particles. As a result, the polymer jet was subjected
to a larger net stretching force, producing bers that are thinner than
those prepared without MSMP particles. Although the ber diameter
did not display monotonous correlation with the polymer blend ratio,
PLAPEO bers were signicantly larger (p b 0.05) than those
electrospun from the single polymer, implying that dilation of free
volume occurred on blending the two polymers. This observation is
 R. Dai, L.-T. Lim / Food Research International 77 (2015) 467475 471

Fig. 3. Scanning electron micrographs of MSMP-containing electrospun ultrane bers: (a) 100PLA0PEO; (b) 75PLA25PEO; (c) 50PLA50PEO; (d) 25PLA75PEO; and (e) 0PLA100PEO. The
histograms show the distribution of ber (y-axis; fraction) in different diameter intervals (x-axis; m).

attributable to different segmental conformations between the dis-
similar PLA and PLA chains, reducing chainchain packing (Wu,
1987). The frequency charts of the ber diameter are presented
along with the SEM micrographs in Figs. 2 and 3, showing the effect
of the polymer blend ratio on the size distribution of the bers. As
shown, thicknesses for the pristine bers tended to be normally dis-
tributed for all polymer blends, but MSMP-containing bers showed
skewed thickness distribution. Also, the addition of PEO to pure PLA
tended to increase the variation in ber thicknesses, as shown in
Fig. 2(b) and (c).
472 R. Dai, L.-T. Lim / Food Research International 77 (2015) 467475
Table 1
Mean ber diameters and standard deviations of pristine and MSMP-containing
electrospun nonwovens.
Fiber membranes Without MSMP, m With MSMP, m
100PLA0PEO 1.09 0.31a 0.81 0.40a
75PLA25PEO 1.68 0.85b 1.46 0.58b
50PLA50PEO 2.02 0.44c 1.50 0.46b
25PLA75PEO 1.75 0.27b 1.48 0.39b
0PLA100PEO 1.31 0.34d 0.77 0.37a
Values are mean standard deviation (n = 200). Numbers within a column with differ-
ent superscript letters indicate statistical signicant difference (p b 0.05). Statistical anal-
ysis was conducted using one-way ANOVA with Tukey's post-hoc test.
3.2. Release of AITC from encapsulated MSMP
The formation of AITC in mustard seed is triggered by the hydrolysis
of a glucosinolate, sinigrin, catalyzed by the cell wall-bound myrosinase
(Delaquis & Mazza, 1995). In this study, the entrapment of MSMP was
achieved by dispersing MSMP grounds in a polymer solution, followed
by electrospinning the suspension into a nonwoven composite. To acti-
vate the release of AITC from the resulting MSMP-containing nonwo-
ven, the polymeric carrier should be moderately hydrophilic to allow
the penetration of water substrate into the entrapped MSMP particles.
The polymer should be soluble in a non-aqueous solvent to prevent pre-
mature hydrolysis of sinigrin during electrospinning. To this end, PLA
was selected since it is highly permeable to water and it is soluble in
chloroform/DMF blend conducive for electrospinning (Huang et al.,
2003; Vega-Lugo & Lim, 2009; Zhou & Lim, 2009). When the MSMP-
containing nonwovens were exposed to moisture, several diffusion-
controlled phenomena were involved in triggering the release of AITC:
(1) sorption of water onto the surface of electrospun bers; (2) diffusion
of water through the polymer matrix; (3) uptake of water by MSMP
particles; (4) hydrolysis of sinigrin forming AITC; (5) diffusion of AITC
through MSMP and ber matrices towards the ber surfaces; and nally
(6) desorption of AITC into the headspace air. Thus, manipulating fac-
tors that change the hydrophilicity and barrier properties of the ber
carrier will allow one to control the release kinetic of AITC. For this pur-
pose, an amphiphilic PEO was added to the polymer solution at different
ratios to plasticize, as well as to adjust the hydrophilicity of the nonwo-
vens. These two polymers share similar solubility parameter () values
in the range of 9.59.8. Accordingly, the two polymers are expected to
form homogeneous blends (Younes & Cohn, 1988).
Representative release proles of AITC from the electrospun nonwo-
vens are presented in Fig. 4. As shown, all plots show a brief initial lag
phase due to the time needed for the humidity to buildup in the head-
space of the test bottle. Beyond the lag phase, AITC released at rates
Fig. 4. Representative cumulative AITC release proles from MSMP-containing
electrospun nonwovens in 1 L glass jar at saturated RH, 20 C. Symbols and continuous
lines are, respectively, experimental data and tted curves based on the modied
Gompertz equation.
dependent on the polymer blend ratio, so as the maximal amounts of
AITC released. To better understand the release kinetic, Eq. (4) was
tted to the release plots. The estimated Qmax, k, and parameters are
summarized in Table 2. The lag times tended to increase with increasing
PEO content, which may be caused by the greater afnity of PEO with
water than PLA, thereby extending the time needed for the water mol-
ecules to reach the enzymatic reaction sites in MSMP particles. Howev-
er, after the initial hydration phase, the release rates of AITC from the
PEO-containing nonwovens were signicantly higher (p b 0.05) than
that of pure PLA, due to increased hydrophilic properties of the ber car-
riers when PEO was added. Similarly, the addition of PEO also enhanced
the total amount of the AITC release. As shown in Table 2, Qmax values
were signicantly higher (p b 0.05) for the blended nonwovens than
that of pure PLA nonwovens. The effect of PEO on enhancing ber hy-
drophilicity when blended with PLA polymers has been reported by
Honarbakhsh and Pourdeyhimi (2011), who observed a signicantly
lower water contact angle (47.4) on electrospun PEOPLA (10:90,
w%) bers than that of neat PLA bers (58.4). The increased hydrophi-
licity will enhance the plasticization effect of water on the ber matri-
ces, facilitating the diffusion of water and AITC molecules. According
to this hypothesis, it is expected that 0PLA100PEO, being the most hy-
drophilic among the nonwovens tested, should exhibit the highest
AITC release rate and amount released. However, data from Table 2
and Fig. 4 do not show this trend. This anomaly can be explained by
the visible shrinkage of the 0PLA100PEO nonwoven at the end of the ex-
periment. The shrinkage of 0PLA100PEO was likely caused by the solu-
bilization of the PEO ber as it absorbed large quantity of water,
reducing the surface area available for moisture and AITC mass trans-
ports. This effect can be seen in Fig. 4, where the slope for 0PLA100PEO
is comparable to those of PLA and blended membranes during the rst
20 h, but the slope noticeably decreased as time progressed due to the
collapsed nonwoven matrices. In comparison, other non-woven speci-
mens did not display visible shrinkage at the end of the experiment.
The sinigrin content of the defatted MSMP, determined using the meth-
od reported in our previous study, was 5.5% w/w (Dai & Lim, 2014).
Complete hydrolysis of this amount of sinigrin would result in
13.7 mg AITC/g MSMP in the 1 L headspace jar, which is greater than
the estimated Qmax values as shown in Table 2. This observation indi-
cates that the maximal amount of AITC released from MSMP particles
encapsulated within the present PLAPEO ber carrier, under the pre-
scribed experimental conditions, would be about 64% (50PLA50PEO)
of the theoretical value. It is noteworthy that the maximal amount of
AITC released will be dependent on temperature, humidity, as well as
the morphology of the electrospun bers. These factors should be
taken into consideration when developing an MSMP-based active pack-
age system for shelf-life extension of food.
3.3. FTIR analyses on pristine and MSMP-containing electrospun ber
membranes
FTIR analysis was conducted to further elucidate the interaction be-
tween different components within the electrospun bers. The spectra
Table 2
Estimated parameters of modied Gompertz equationQmax, k and lag time constants of
AITC release proles for MSMP-containing electrospun nonwoven at 20 C and 100% RH.
MSMP-containing nonwovens Qmax Rate k Lag time
mg/g MSMP1 L h1 h
100PLA0PEO 4.59 0.33a 0.09 0.00a 5.20 0.33a
75PLA25PEO 7.29 0.16b 0.30 0.02b 4.86 0.63a
50PLA50PEO 8.88 0.05c 0.38 0.01c 6.24 1.33a
25PLA75PEO 8.37 0.30c 0.25 0.03d 9.50 0.90a
0PLA100PEO 7.40 1.48b 0.25 0.01d 8.41 2.41a
Values are mean standard deviation (n = 3). Means within a column with different su-
perscript letters indicate statistical signicant difference (p b 0.05). Statistical analysis was
conducted using one-way ANOVA with Tukey's post-hoc test.
 R. Dai, L.-T. Lim / Food Research International 77 (2015) 467475 473

of electrospun pristine and MSMP-containing nonwovens, are present-

ed in Figs. 5 and 6, respectively. As an aliphatic polyester, PLA contains
exible ester bonds in the backbone structure with the characteristic
carbonyl (C_O) absorption bands at 1751 cm1, whereas the C\\O\\C
stretching bands of PLA appeared at 1082 cm1. Absorption bands that
relate to the amorphous and crystalline structure of PLA appeared at
862 and 755 cm 1, respectively (Auras, Lim, Selke, & Tsuji, 2010;
Kang et al., 2001; Kister, Cassanas, & Vert, 1998; Xu et al., 2009;
Younes & Cohn, 1988). PEO is a polymer with the molecular formula
(\\CH2CH2O\\)n. The characteristic C\\O\\C absorbance band is locat-
ed at 1097 cm1. The bands detected at 962 and 843 cm1 have been
attributed to the rocking and twisting of C\\H2 and stretching of
C\\H. Both of these peaks have been associated with the crystalline
structure of PEO (Oliveira et al., 2013; Pielichowski & Flejtuch, 2005;
Wen et al., 1996; Younes & Cohn, 1988; Zhou, Chu, Wu, & Wu, 2011).
In Fig. 5, similar spectral features are observed between electrospun
PEO bers and as-received PEO powders, indicating that the
electrospinning process did not alter the molecular conformation of
the polymer (Fig. 5). In the polymer blends, the C\\O\\C stretching
band of PEO at 1097 cm 1 overlapped with the C\\O\\C band from
PLA at 1082 cm 1. No signicant shift was observed for these two Fig. 6. Comparison of FTIR spectra of MSMP-containing electrospun nonwovens.

peaks as the polymer blend proportion changed. Similarly, the C\\H

stretch at 2880 cm1 from the PEO backbone remained unchanged at
all polymer blend ratios. A similar observation was reported by
Younes and Cohn (1988) in their extruded PLAPEG blend lms, who
attributed the lack of interaction to the low concentration of\\OH end
groups and the absence of other moieties capable of developing the in-
termolecular interactions.
For the MSMP-containing bers (Fig. 6), spectra for all polymer sam-
ples displayed an extra amide I band at 1649 cm1, attributable to the
protein fraction present in MSMP that contained 40.5% protein (Dai &
Lim, 2014). Comparing Figs. 5 and 6, vibration frequencies for C\\O\\C
(1097 cm 1) and C_O (1751 cm 1) of PEO and PLA, respectively,
remained unchanged after the incorporation of MSMP. In summary,
the FTIR analysis did not reveal any specic interaction between
MSMP and the polymers.
3.4. Thermal analyses
Thermograms for electrospun pristine PLA, PEO and PLAPEO non-
wovens are presented in Fig. 7. The endothermic melting peaks of
0PLA100PEO and 100PLA0PEO electrospun bers were 72 C and
168 C, respectively, comparable with the values previously reported
in the literature (Coleman, Yang, & Painter, 1992; Nijenhuis, Colstee,
Grijpma, & Pennings, 1996). PLA and PEO are semi-crystalline polymers,
the crystallinity of which can be estimated from their melting enthalpy
(Nijenhuis et al., 1996; Oliveira et al., 2013; Younes & Cohn, 1988). An
attempt was done to study the miscibility of PLA and PEO by evaluating
the Tg values of the polymer blends. Unfortunately, a considerable over-
lap existed between the Tg of PLA and Tm of PEO, making quantitative
measurement difcult. Furthermore, an endothermic relaxation peak
that appeared near the glass transition of PLA (Lim et al., 2008) further
complicated the analysis. Nevertheless, a decreasing trend of Tg of PLA
with increasing PEO content is visible in Fig. 7, indicating that the
added PEO acted as a plasticizer to PLA. The plasticizing effect of PEG
(b50%, w/w) on extruded PLA has been reported by Sheth, Kumar,
Dave, Gross, and McCarthy (1997).
The melting peak temperature of pure PEO shifted from 72 C to
lower temperatures when the weight ratio of PLA polymer solution
increased from 0 to 50%. This shift is possibly due to the disruption of
PEO chain packing by the PLA polymer, producing imperfect crystalline
structures, leading to a lower melting temperature for PEO. On the other
hand, the Tm of PLA remained unchanged for all polymer blends regard-
less of the PEO ratio, indicating that the crystalline phase of PLA was not
affected by the incorporation of PEO (Fig. 7). Similar phenomena were
reported by Younes and Cohn (1988) for their PLAPEG cast lms.

Fig. 5. Comparison of FTIR spectra of the pristine electrospun nonwovens. Fig. 7. DSC thermograms of pristine electrospun nonwovens.
474 R. Dai, L.-T. Lim / Food Research International 77 (2015) 467475
The thermogram for MSMP alone showed a weak endothermic peak
at about 73 C and a relatively strong endotherm peak at about 193 C
(Fig. 8). The MSMP matrix is complex and thus the origins for these
thermal events are unknown, but they are likely due to denaturation
of proteins (Byczynska & Barciszewski, 1999; Marcone et al., 1997)
and thermal degradation reactions as the MSMP samples were heated
to elevated temperatures. The protein content in the MSMP samples
was 40.5 0.6% (Dai & Lim, 2014). When the MSMP particles were dis-
persed in the bers, both endothermic peaks shifted to higher temper-
atures (Fig. 8). This observation could be caused by the MSMP
polymer interaction and/or leaching of soluble MSMP-components
into the polymer solution that enhanced the thermal stability. On the
other hand, the addition of MSMP into the polymer resulted in signi-
cant Tm lowering for 100PLA0PEO and 0PLA100PEO, from 168 to
162 C and from 72 to 62 C, respectively (Fig. 8). These shifts suggest
that MSMP interfered with the chain packing of the crystalline phases
for PLA and PEO during the electrospinning processes. The Tm lowering
effect may be a result of the interaction of MSMP components, such as
starch and cellulose, with the polymers. For example, starch has been
reported to depress Tm of PEO, due to the hydrogen bonding interaction
between hydroxyl groups of starch and ether oxygen from the repeated
units of PEO (Pereira, Gouveia, Carvalho, Rubira, & Muniz, 2009). In ad-
dition, the Tm depression effect of cellulose on electrospun PLA has been
reported in the literature (Shi et al., 2012). The contribution of lipids to
the Tm lowering effect will be negligible since the MSMP particles used
in the present study were defatted (0.00% lipid).
4. Conclusions
This study evaluated the feasibility of encapsulating MSMP particles
in PLAPEO nonwovens using an electrospinning technique. Diameters
of PLAPEO blend bers were PLAPEO blend ratio dependent, with
overall smaller diameters when MSMP particles were incorporated. De-
spite the larger MSMP particles than the electrospun bers, the surface
of MSMP was fully covered by the polymer, as revealed by SEM analysis.
The rate and quantity of AITC released could be manipulated by modify-
ing the polymer composition of the electrospun bers. FTIR analyses did
not reveal any specic interaction between the IR active bonds in PLA,
PEO and MSMP. However, DSC analysis showed that when MSMP was
incorporated into the polymers, signicant decreases in PLA and PEO
melting temperatures were observed, indicating that components
from MSMP had interfered with the chain packing of the crystalline
phases of the polymers. In summary, this study showed that the
electrospun PLAPEO nonwoven is a promising carrier for controlling
the release of AITC from MSMP. In view of the well-established
Fig. 8. DSC thermograms of MSMP-containing electrospun nonwovens.
antimicrobial efcacy of AITC in many food systems, further evaluation
of the MSMP-containing nonwovens will be needed in order to eluci-
date their feasibility in active food packaging applications.
Acknowledgments
This research was funded by the Natural Sciences and Engineering
Research Council (NSERC) of Canada, and supported by Biofume Tech-
nologies Inc.
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