Microencapsulation of ax oil with zein using spray and freeze drying

Scrates Quispe-Condori
a, b
, Marleny D.A. Saldaa
a,
*
, Feral Temelli
a
a
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, T6G 2P5, Canada
b
Escuela de Ingeniera de Alimentos, Universidad Peruana Unin, Carretera Central Km. 19.5, aa, Lima, Peru
a r t i c l e i n f o
Article history:
Received 16 June 2010
Received in revised form
29 December 2010
Accepted 8 January 2011
Keywords:
Flax oil
Freeze drying
Microencapsulation
Response surface design
Spray drying
Zein
a b s t r a c t
Microencapsulation of ax oil was investigated using zein as the coating material. Central Composite
Design e Face Centered was used to optimize the microencapsulation with respect to zein concentration
(x
1
) and ax oil concentration (x
2
) using spray drying. Also, freeze drying was carried out at two zein:oil
ratios. The quality of microcapsules was evaluated by determining encapsulation efciency, owing
properties (Hausner ratio), and evaluating the morphology with scanning electron microscopy. The
response surface model for microencapsulation efciency showed a high coefcient of determination
(R
2
0.992) and a non-signicant lack of t (p 0.256). The maximum microencapsulation efciencies
were 93.26 0.95 and 59.63 0.36% for spray drying and freeze drying, respectively. However,
microcapsules prepared by spray and freeze drying had very poor handling properties based on the
Hausner ratio. The bulk density decreased with an increase in zein concentration at the same ax oil
concentration. The morphology of the ax oil microcapsules depended on the zein:ax oil ratio and the
process used for microencapsulation. Flax oil microcapsules prepared by spray drying appeared to be
composed of heterogeneous spheres of various sizes at high zein:ax oil ratios. Microcapsules prepared
by freeze drying resulted in agglomerated small spheres. These microcapsules might nd a niche as
functional food ingredients.
2011 Elsevier Ltd. All rights reserved.
1. Introduction
There has been a growing interest in research, development and
commercialization of functional food ingredients over the past
decade (Day, Seymour, Pitts, Konczak, & Lundin, 2009; Shahidi,
2009). The addition of u-3 and u-6 polyunsaturated fatty acids
(PUFA) to functional food ingredients and their consumption in
dietary supplements have experienced signicant increases
(OBrien, 2009; Sanguansri & Augustin, 2007). These fatty acids
have been associated with a variety of health benets, such as
reducing the risk of coronary heart diseases, hypertension, arthritis,
and immune response disorders (Rubio-Rodrguez et al., 2010). Fish
oil, ax oil, and more recently, algae oil, are the most commonly
used sources of u-3 fatty acids most commonly used (Rubio-
Rodrguez et al., 2010). However, one of the major drawbacks of
oils containing a high amount of PUFA is their rapid oxidation,
which involves the formation of toxic products such as peroxides or
undesirable off-avor compounds (OBrien, 2009).
The use of encapsulation technologies to retard or avoid the
oxidation of these oils has been reported and it has drawn
considerable attention in the food industry (Ahn, Kim, Lee et al.,
2008; Ahn, Kim Seo et al., 2008; Klaypradit & Huang, 2008;
Klinkesorn & McClements, 2009; Kolanowski, Laufenberg, & Kunz,
2009; Omar, Shan, Zou, Song, & Wang, 2009; Partanen, Yoshii,
Kallio, Yang, & Forssell, 2002; R, 1998; Sanguansri & Augustin,
2007; Velasco, Marmesat, Dobarganes, & Mrquez-Ruiz, 2006).
Some of the commonly used coating materials for the microen-
capsulation of u-3 fatty acids are proteins, lipids, polysaccharide
gums and cellulose (Sanguansri & Augustin, 2007). However, the
selection of an adequate coating material for PUFA depends on its
capacity to stabilize and protect the oil from degradation during
processing and storage conditions, and it also has to be approved
for food use. Zein, the prolamin fraction of corn protein (Shukla and
Cheryan, 2001), has long been recognized for its coating ability for
the encapsulation of bioactive compounds (Hurtado-Lpez &
Murdan, 2006; Jnsdttir, Bragadttir, & Arnarson, 2005; Liu, Sun,
Wang, Zhang, & Wang, 2005; Parris, Cooke, & Hicks, 2005; Zhong
& Jin, 2009; Zhong, Jin, Xiao, Tian, & Zhang, 2008) and sh oil
(Zhong, Tian, & Zivanovic, 2009). For lipid compounds, zein has
been shown to adsorb fatty acids and produce zein sheets (Padua &
Wang, 2009). Zein can also bind and enrobe lipids, keeping them
from deteriorative changes. In addition, the antioxidant activity of
zein has been studied in powder systems (Matsumura, Andonova,
Hayashi, Murakami, & Mori, 1994; Wang, Fujimoto, Miyazawa, &
Endo, 1991; Wang, Miyazawa, & Fujimoto, 1991).
* Corresponding author. Tel.: 1 780 492 8018; fax: 1 780 492 8914.
E-mail address: marleny@ualberta.ca (M.D.A. Saldaa).
Contents lists available at ScienceDirect
LWT - Food Science and Technology
j ournal homepage: www. el sevi er. com/ l ocat e/ l wt
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doi:10.1016/j.lwt.2011.01.005
LWT - Food Science and Technology 44 (2011) 1880e1887
Flax oil is a good source of PUFA, especially linolenic acid (C18:3
u-3) (Bozan &Temelli, 2002). However, this oil has lowstability and
high susceptibility to oxidation. Encapsulation of ax oil using zein
as a coating material has not been reported in the literature.
Therefore, the objective of this study was to evaluate the ability of
zein to act as a microencapsulating agent for ax oil using spray and
freeze drying, considering that Canada is the worlds largest ax
producer. The effects of the concentrations of zein and ax oil were
studied using Response Surface Methodology, Central Composite
Design e Face Centered, in order to optimize the formulation to
produce ax oil microcapsules. The microcapsules were analyzed
for their microencapsulation efciency, owing properties and
morphological characteristics.
2. Material and methods
2.1. Materials
Cold pressed and unrened ax oil was obtained from a local
market, Gold Top Organics (Edmonton, AB, Canada). Corn zein (NF
F4400C Non-GMO/IP grade) was supplied by Freeman Industries
LLC (Tuckahoe, NY, USA). Food grade anhydrous ethyl alcohol was
purchased from Commercial Alcohols Inc. (Toronto, ON, Canada).
Petroleum ether and hexane were acquired from Fisher Scientic
(Ottawa, ON, Canada).
2.1.1. Fatty acid composition
The fatty acid (FA) composition of ax oil was determined using
a modied fatty acid methyl esters (FAME) method. The FAME was
analyzed using a gas chromatograph with a ame ionization
detector (GC-FID) (Varian Model 3400, Agilent Technologies, Mis-
sissauga, ON, Canada) equipped with a fused silica capillary column
(60 m0.32 mm, DB-5, J&WScientic, Folsom, CA, USA). The carrier
gas was helium. The temperatures of the injector and detector were
235 and 250

C, respectively. The column was heated to 70

C and
held for 0.2 min, programmed to increase at a rate of 35

C/min to
210

C, then at a rate of 2

C/min to 235

C and held for 8.3 min.
The ax oil dissolved in hexane (100 mL) was transmethylated to
FAME by heating at 50

C for 15 min with a mixture of 0.5 mol/L
sodium methoxide (80 mL) and methyl acetate (40 mL). Methyl
heptadecanoate was used as an internal standard at the concen-
tration of 1 mg/mL. One milliliter of the standard solution was
added to the mixture. To separate the FAME, water (2 mL) and 2 mL
of hexane were added to the mixture and the solution was centri-
fuged (GLC-2B Sorvall Dupont Instruments, Wilmington, NC, USA)
at 1237 g for 1 min. The organic phase was separated and 1 mL of
the sample was injected to the gas chromatograph.
2.2. Experimental design
Central Composite Design e Face Centered (CCDeFC) was used
to investigate the microencapsulation efciency with respect to
zein and ax oil concentrations.
Ten experimental settings, consisting of 2
2
factorial points, 4 star
points and 2 central points were generated with 2 factors and 2
levels using the software STATISTICA 7 (Stat Soft Inc., Tulsa, OK,
USA). The units, the actual and coded levels of the independent
variables are presented in Table 1. For this design, the following
quadratic model was used:
y b
0
b
1
x
1
b
2
x
2
b
11
x
2
1
b
22
x
2
2
b
12
x
1
x
2
e
(1)
where y is the dependent variable predicted by the model
(response); b
0
is the constant coefcient of intercept; b
1
and b
2
are
the linear effects, b
11
and b
22
are the quadratic effects, b
12
is the
linear-by-linear interaction, and e is the error term. Zein and ax oil
concentrations are represented by x
1
and x
2,
respectively, as the
coded independent variables. The responses measured were
moisture content, particle yield, microencapsulation efciency and
owing properties.
2.3. Encapsulation of ax oil
Microcapsules of ax oil were prepared using two different
processes: (i) Spray drying, and (ii) Freeze drying.
2.3.1. Spray drying
Corn zein was dissolved in 150 mL of aqueous ethanol solution
(85 mL ethanol/100 mL) according to the concentrations reported
in Table 1. The mixture was stirred at 2500 rpm for 1 min using an
homogenizer (Model L4RT-W, Silverson Machines, Chesham Bucks,
England). Flax oil was added to form an emulsion and the
homogenization was maintained at this rate for 4 min. This level of
mixing was found to be sufcient, conrming the ndings of Chen
and Tao (2005) as the emulsion was stable even after a month (data
not shown). The emulsion was then spray-dried using a B-190 mini
spray drier (Bchi Labortechnik AG, Flawil, Switzerland) at the ow
rate of 9 mL/min. The inlet (controlled) and outlet (measured) air
temperatures were 135 and 55e60

C, respectively. The spray-dried

microcapsules collected in the cyclone were recovered and stored
at 17

C until further analysis.
2.3.2. Freeze drying
The freeze-dried microcapsules were produced according to the
protocol described by Parris et al. (2005) with modications.
Emulsions were prepared as described above and were rapidly
mixed using the homogenizer with 350 mL of distilled water. The
solution was then freeze-dried overnight using a lyophilizer (Virtis
Co., Gardiner, NY, USA). The freeze-dried microcapsules were
recovered and stored at 17

C until further analysis. Only two
formulations corresponding to Runs #1 and #3 under spray drying
were tested using freeze drying since the other formulations
resulted in sticky mixtures/powders, which were difcult to
process with the freeze-drier. Due to the limitations of the freeze-
drier in terms of the amount of ethanol that can be handled, extra
water was added to these mixtures during the preparation of
Table 1
Central composite design e Face Centered: coded and actual factor levels and
experimental matrix for the optimization of the ax oil microencapsulation.
Factors Coded level (x
i
)
1 0 1
x
1
: Zein concentration (g/100 mL) 6 10 14
x
2
: Oil concentration (g/100 mL) 0.5 1 1.5
Run Coded value
(Actual value)
Zein:Oil
Ratio
x
1
(x
1
) x
2
(x
2
)
1 1 (6) 1 (0.5) 12:1
2 1 (14) 1 (0.5) 28:1
3 1 (6) 1 (1.5) 4:1
4 1 (14) 1 (1.5) 9.3:1
5 1 (6) 0 (1) 6:1
6 1 (14) 0 (1) 14:1
7 0 (10) 1 (0.5) 20:1
8 0 (10) 1 (1.5) 6.7:1
9 (c) 0 (10) 0 (1) 10:1
10 (c) 0 (10) 0 (1) 10:1
c central point.
S. Quispe-Condori et al. / LWT - Food Science and Technology 44 (2011) 1880e1887 1881
emulsions (as also suggested by Parris et al. (2005)), which
contributed to the formation of sticky mixtures/powders.
2.4. Moisture content
The moisture content of each sample (500 mg) was determined
by loss of weight in an infrared moisture analyzer (Sartorius, Model
MA45, Elk Grove, IL, USA) at 130

C. Each analysis was repeated
three times.
2.5. Particle yield
The particle yield for each experimental assay was calculated
using Eq. (2) as reported by Zhong and Jin (2009):
Particle yield
Mass of collected product
Non solvent mass in the feed
100 (2)
2.6. Microencapsulation efciency (MEE)
The microencapsulation efciency is dened as the ratio of core
material in the nal dried microcapsules to that in the original
emulsion (Zilberboim, Kopelman, & Talmon, 1986) and was calcu-
lated according to Eq. (3), as provided elsewhere (Ahn, Kim, Lee
et al., 2008; Ahn, Kim, Seo et al., 2008; Omar et al., 2009; Pauletti
& Amestoy, 1999; Velasco et al., 2006).
MEE%
Total oil Surface oil
Total oil
100 (3)
2.6.1. Total oil
An aliquot of 500 mg of each ax oil microcapsule treatment
was dissolved in 5 mL of aqueous ethanol solution (85 mL ethanol/
100 mL). The samples were stirred gently and placed in a sonicator
bath (Branson Ultrasonic Corporation, Danbury, CT, USA) at room
temperature. After 15 min, the samples were extracted ve times
using a total of 25 mL of petroleum ether, as follows: Each sample
was mixed gently with 5 mL of petroleum ether. After 1 h of
contact, phase separation was observed and the upper phase was
poured into a tared watch glass. The ethanol solution was re-
extracted using an additional 5 mL of petroleum ether. The same
procedure was repeated three more times. Then, the solvent was
evaporated from the tared watch glass at room temperature, and
the extracted oil was dried until constant weight was achieved.
The cooled watch glass was weighed and the total oil content was
calculated. Triplicate determinations were performed for each
sample.
2.6.2. Surface oil
The surface oil, also known as the non-encapsulated oil fraction,
was determined according to Velasco et al. (2006) with modica-
tions. A sample (500 mg) was weighed in an extraction thimble of
lter paper (Whatman No 40) prepared manually. The extraction
thimble was placed in a tared 25 mL glass bottle. The microcapsules
were washed 5 times using 1 mL of petroleum ether at room
temperature for a total of 5 mL. After ltration through the lter
paper, the solvent was evaporated from the glass bottle and the
extracted oil was determined gravimetrically, which was consid-
ered as the surface oil. Triplicate determinations were performed
for each sample.
2.7. Flowing properties
The bulk (r
B
) and tapped (r
T
) densities were determined in
a 25 mL glass graduated cylinder as described by Chinta et al.
(2009). Three grams (3 g) of each sample (m
0
) was poured
through a funnel into the cylinder. Then, the cylinder was slightly
tapped to collect the powder sticking to the wall of the cylinder. The
volume (V
0
) was read directly from the cylinder and used to
calculate the bulk density (r
B
m
0
/V
0
). For tapped density (r
T
m
0
/
V
n
), the cylinder was tapped until a constant volume (V
n
) was
reached.
The powder owability was evaluated using the Carrs Index or
percent compressibility (C) and the Hausner Ratio (HR) (Turchiuli
et al., 2005). The Carrs Index and the Hausner Ratio were calculated
using the following equations, respectively:
C
r
T
r
B
r
T
100 (4)
HR
r
T
r
B
(5)
2.8. Morphological analysis
Morphology and surface appearance of zein and microcapsules
were examined using a Scanning Electron Microscope (SEM). Each
sample was glued onto a black adhesive tape, mounted on a stain-
less steel stub and subjected to critical-point drying (Bal-Tec CPD
030; Bal-Tec AG, Balzers, Liechtenstein). After drying, samples were
examined with a scanning electron microscope (EVO MA15, Carl
Zeiss, Oberkochen, Germany) operated at 10 kV.
2.9. Statistical analysis
The adequacy of the response surface model was determined
using the regression coefcient analysis (R
2
) and the Analysis of
Variance (ANOVA) with the lack of t test. ANOVA was performed
to assess the signicance of the effects of the independent variables
on response variables and the statistical model. For the statistical
analysis of the regression, the total error term was separated into
pure error and the error of the lack of t of the model, which
represents a goodness of t test for response surface models
(Montgomery, 2006).
The determination of the moisture content, microencapsulation
efciency and owing properties were performed in triplicate and
the means were compared using the Tukey test, at a 95% condence
interval.
3. Results and discussion
3.1. Fatty acid composition of feed ax oil
Linolenic acid (C18:3 u-3) was the major fatty acid (59.85
0.39 g/100 g), followed by oleic (C18:1) (16.04 0.21 g/100 g),
linoleic (C18:2) (15.86 0.06 g/100 g), palmitic (C16:0)
(5.17 0.03 g/100 g) and stearic (C18:0) (3.07 0.08 g/100 g)
acids. The fatty acid composition of the ax oil was in agreement
with the proles previously reported by Bozan and Temelli (2002).
The stability of the encapsulated ax oil and the changes in fatty
acid composition over storage are the subject of another report.
There was no change in the u-3 fatty acid content due to the
encapsulation process for spray-dried particles. But, the content of
u-3 fatty acid decreased for freeze-dried particles as the surface oil
content was high.
S. Quispe-Condori et al. / LWT - Food Science and Technology 44 (2011) 1880e1887 1882
3.2. Moisture content, particle yield and microencapsulation
efciency
The characteristics of the dry microcapsules depend on the
physical and chemical properties of the feed, the dryer design, and
the operating conditions (R, 1998). In this study, the operating
conditions of temperature and ow rate in the spray dryer were
kept constant.
Moisture content, along with temperature, affects the shelf life
of dried microcapsules. In general, dry foods with moisture
contents between 3 and 10 g/100 g have a good stability during
storage (Klaypradit & Huang, 2008).
Table 2 shows the moisture content of spray- and freeze-dried
microcapsules. The moisture content of zein powder was
4.48 0.04 g/100 g, while that of spray-dried microcapsules was in
the range of 3.49 0.26e5.06 0.14 g/100 g. The moisture content
decreased (p < 0.05) with an increase in ax oil at the same zein
concentration. Increased oil content reduces water activity. The
moisture content of the microcapsules produced by freeze drying
was similar to that of spray-dried powders at a zein:oil ratio of 12:1
(Run #1) but signicantly higher (p <0.05) at the ratio of 4:1. In this
case, the freeze-dried particles have higher moisture content due to
the addition of extra water during the emulsion preparation step.
These samples were dried for two days and obviously a longer
drying time could have reduced the moisture content further.
In Table 3, the ANOVA results are presented for the microen-
capsulation after spray drying. The linear effect of the ax oil
concentration on the moisture content was signicant (p 0.011).
The regression coefcient for Eq (1) showed that the quadratic
model ts the data reasonably well (R
2
0.861).
Particle yield of the microcapsules from different formulations is
presented in Table 2. The particle yield increased (p 0.025) with
zein concentration. Possibly, the presence of a larger amount of zein
may have allowed for the formation of thicker microparticle walls.
The particle yield also varied signicantly (p < 0.05) depending
on the microencapsulation process. Lower particle yields were
obtained by spray drying than by freeze drying using the same
zein:ax oil ratio (Table 2). A similar behavior was observed
previously by Tan, Chan, and Heng (2005) and Parris et al. (2005).
Tan et al. (2005) obtained particle yields in the range of
43.77e68.60 g/100 g for the microencapsulation of modied
starch marine oil (0.7:1e2:1 ratio), results that are comparable to
those obtained in this study. Parris et al. (2005) also obtained high
particle yields in the microencapsulation of essential oil zein by
freeze drying (65e75 g/100 g). In addition, Zhong and Jin (2009)
obtained low particle yields in the microencapsulation of
lysozyme zein by spray drying (32.7e48.7 g/100 g).
The ANOVA results in Table 3 conrmed the signicant effect of
zein concentration x
1
(p 0.025) compared to ax oil concentration
x
2
(p 0.056) on the particle yield. The constants of the regression
equation for the particle yield are presented in Table 4. The lack of
t of the model (p 0.02) for particle yield was reected in the
relatively low regression coefcient (R
2
0.67). This was also
conrmed by the scatter in the data when predicted values versus
observed values were plotted.
The response surface contour plot for particle yield is presented
in Fig. 1(a), which shows that the particle yield could be maximized
at high levels of zein concentration. Furthermore, intermediate
zein:ax oil ratios produce the highest particle yields.
To evaluate the parameters affecting the microencapsulation
efciency, the amounts of surface and total oil of the microcapsules
were rst determined (Fig. 2). The surface oil is critical to micro-
capsule stability, since oil on the surface of a particle may oxidize
more readily in the presence of oxygen. The amount of surface oil
was low (less than 2 g oil/100 g) at high zein:ax oil ratios. The
microcapsules had more surface oil at lowzein:ax oil ratios, and it
increased drastically with a decrease in the ratio below 10:1. These
results demonstrate the importance of having sufcient quantities
of the coating material to encapsulate the oil rather than having
a portion of the oil left on the surface. The same behavior was
Table 2
Moisture content, particle yield, and microencapsulation efciency of ax oil-zein
microcapsules.
Run Moisture
content (g/100 g)
a
Particle yield
(g/100 g)
Microencapsulation
efciency (%)
a
Spray drying
1 5.06 0.14 65.80 89.61 0.51
2 4.92 0.07 78.97 90.57 0.76
3 3.49 0.26 64.22 78.72 0.63
4 4.18 0.23 66.09 75.42 0.62
5 3.88 0.13 68.94 84.45 0.89
6 4.47 0.25 75.07 84.14 0.81
7 4.43 0.07 78.76 93.26 0.95
8 3.55 0.02 77.19 82.40 0.77
9 4.51 0.35 76.55 89.71 0.97
10 4.39 0.21 76.40 89.26 0.82
Freeze drying
1 5.33 0.33 70.50 59.63 0.36
3 4.94 0.05 77.80 32.68 0.46
a
Mean standard deviation based on triplicate determinations.
Table 3
Analysis of variance (ANOVA)
a
of the independent variables for the moisture
content, particle yield and microencapsulation efciency of ax oil with zein after
spray drying.
Factor df Moisture content Particle yield Microencapsulation
efciency
SS p SS p SS p
Linear
x
1
1 0.2492 0.1919
ns
74.6354 0.0246* 1.1618 0.2377
ns
x
2
1 2.0188 0.0111* 42.7263 0.0564
ns
226.9678 0.00004**
Quadratic
x
1
2
1 0.0438 0.5469
ns
120.0761 0.0112* 50.5421 0.0008**
x
2
2
1 0.0098 0.7717
ns
3.3852 0.4958
ns
2.9154 0.0929
ns
Interaction
x
1
x
2
1 0.1998 0.2329
ns
31.8752 0.0832
ns
4.5520 0.0516
ns
Error 1 0.4053 e 24.1703 e 2.4157 e
Total SS 9 2.9210 e 307.3083 e 294.2441 e
R
2a
0.861 0.921 0.992
**p 0.01; *p 0.05; ns: Non-signicant; df: Degree of freedom; SS: Sum of
squares.
a
SS Residual term used for ANOVA.
Table 4
Regression coefcients
a
calculated for the particle yield and microencapsulation
efciency of ax oil with zein by spray drying.
Factor Regression coefcient (b)
Particle yield Microencapsulation efciency
Constant (b
0
) 77.2234 0.0536 88.6572 0.1570
Linear
b
1
3.5269 0.0437 ns
b
2
ns 6.1504 0.1282
Quadratic
b
11
7.3744 0.0692 4.8404 0.2027
b
22
ns ns
Interaction
b
12
ns ns
R
2
0.668 0.962
Lack of t 0.0199* 0.1761
ns
ns: Non-signicant.
a
SS Residual term used in the determination of regression coefcients.
S. Quispe-Condori et al. / LWT - Food Science and Technology 44 (2011) 1880e1887 1883
observed for the freeze-dried microcapsules, where the surface oil
content was reduced in the same proportion. The microencapsu-
lation efciency was in the range of 75.42 0.62% to 93.26 0.95%
for the spray-dried microcapsules (Table 2).
ANOVA was performed to assess the signicance of the inde-
pendent variables for the MEE (Table 3). It was conrmed that the
ax oil concentration had more inuence than the zein concen-
tration on the microencapsulation efciency. The regression coef-
cient for the MEE equation (R
2
0.992) (Table 3) showed that the
quadratic model tted the experimental data adequately. A plot of
the expected normal values versus residuals showed that the data
are very close to the straight line with little scatter, indicating that
the surface model is good.
The tted mathematical model does not show a signicant lack
of t (p 0.176), conrming the adequacy of the statistical model
calculated for the microencapsulation efciency. The analysis of the
regression coefcients (Table 4) conrmed that the linear co-
efcient for the ax oil concentration x
2
and the quadratic coef-
cient for the zein concentration x
1
2
have a signicant inuence on
the regression equation for the microencapsulation efciency
Fig. 1. Estimated contour plot in the central composite design experiment obtained for: (a) particle yield, (b) microencapsulation efciency, and (c) bulk density as a function of the
ax oil and zein concentrations.
Fig. 2. Surface and total oil contents of ax oil encapsulated in zein by spray drying
(SD) and freeze drying (FD). Means of triplicate with standard deviations presented in
the error bars (D SD Surface oil, : FD Surface oil, , SD Total oil, - FD Total
oil).
S. Quispe-Condori et al. / LWT - Food Science and Technology 44 (2011) 1880e1887 1884
(p <0.05). As shown in Table 2, an increase in ax oil concentration
resulted in low microencapsulation efciency. This behavior is
conrmed by the negative sign of the linear regression coefcient
for ax oil (Table 4). Similar results were presented by Omar et al.
(2009) for the microencapsulation of ax oil using gum Arabic.
For the studied experimental conditions, the inuence of zein
concentration was not signicant, unless its value increased in
quadratic form.
In agreement with the maximum experimental microencapsu-
lation efciency obtained in Run #7 (93.26 0.95%) (Table 2),
a predicted MEE value of 94.25% was obtained by using the
response surface model. This result conrms that the quadratic
model could be used to predict the MEE within the levels of the
parameters studied by the experimental design.
In order to determine if the stationary point is a maximum,
minimum or saddle, the response surface was characterized using
an indirect and canonical analysis. The indirect analysis examines
the tridimensional plot of the response with its corresponding
bidimensional contour plot, while the canonical analysis interprets
the geometry of the estimated surface in the region of experi-
mentation (Montgomery, 2006). Fig. 1(b) shows the contour plot
for the estimated microencapsulation efciency in terms of the
interactions between the ax oil and zein concentrations. The
response surface contour plot clearly showed that the microen-
capsulation efciency could be maximized at lower values of ax oil
concentration and intermediate level of zein concentration (20:1
ratio).
From the canonical analysis, it was determined that the coef-
cients of the model have negative signs (data not shown), indi-
cating that the stationary point is the maximum. Low oil
concentration generally results in high encapsulation efciency and
therefore lowsurface oil content. As presented in the literature, the
oil concentration and coating material are critical factors for the
microencapsulation process (Ahn, Kim, Lee et al., 2008).
3.3. Flowing properties
In general, the microencapsulation efciency is used as a quality
parameter for the dried microcapsules. However, other quality
control parameters are bulk density, Carr Index and Hausner Ratio
to assess powder owability (Fitzpatrick, 2005).
As known, a dry product with a high bulk density can be stored
in small containers compared to a product with low bulk density.
The Hausner ratio and the Carr index can be used to determine
powder properties during processing and storage conditions. In
addition, the Hausner ratio, Carr index, angle of repose, and
compression test provide information on powder owability
(Barbosa-Cnovas & Juliano, 2005).
The owing properties for ax oil microcapsules and zein
powder are listed in Table 5. According to the classication pre-
sented in Table 6 for powder owability, zein has excellent owing
properties. However, microcapsules prepared by spray- and freeze-
drying had poor or very poor handling properties. The microcap-
sules prepared in this study were very compressible compared to
other oil microcapsules prepared by spray drying as reported in the
literature (Fuchs et al., 2006; Onwulata, Konstance, & Holsinger,
1996; Turchiuli et al., 2005). The higher Hausner ratio means that
the powder is more cohesive and less able to ow freely.
It was observed that a small increase in the Hausner ratio and
Carr index due to an increase in zein concentration resulted in low
powder owability. However, the Tukey test showed no real change
in the powder owability (p > 0.05). The same result was observed
for microcapsules produced by spray or freeze drying at similar
zein:oil ratios, despite the decrease of the Hausner ratio and Carr
index. Turchiuli et al. (2005) and Fuchs et al. (2006) reported
similar results of poor owability for oil microencapsulated by
a mixture of maltodextrin and acacia gum.
The bulk densities found in this study were typical of encap-
sulated powders (Onwulata et al., 1996). A few exceptions, with
high bulk densities, are microcapsules of a mixture of oil acacia
gum (Fuchs et al., 2006) and butteroil modied corn starch
(Onwulata et al., 1996). For spray-dried microcapsules, it was
observed that microcapsules produced at low zein:ax oil ratio are
denser (p < 0.05) than microcapsules at high zein:ax oil ratios.
Therefore, the bulk density is inversely related to an increase in the
zein:ax oil ratio, mainly inuenced by the zein concentration as
demonstrated by the Tukey test result. For the same experimental
condition, there was no signicant difference (p > 0.05) between
the densities of microcapsules prepared by spray or freeze drying.
Table 5
Flowing properties of ax oil-zein microcapsules prepared by spray and freeze
drying.
Run Flowing properties
Bulk density (kg/m
3
)
a
Hausner ratio
a
Carrs index (C)
a
Spray drying
1 272.85 30.66 1.68 0.15 40.14 5.07
2 174.13 7.58 1.70 0.0 41.02 2.24
3 282.14 6.19 1.53 0.09 34.40 3.82
4 240.74 16.04 1.71 0.13 41.13 4.80
5 279.37 11.00 1.51 0.07 33.72 3.31
6 192.81 18.71 1.96 0.17 48.65 4.26
7 219.44 7.37 1.69 0.08 40.82 2.62
8 251.18 16.04 1.77 0.12 43.47 3.79
9 263.03 21.06 1.61 0.11 37.82 4.20
10 227.71 14.55 1.61 0.07 37.68 2.77
Freeze drying
1 309.18 24.11 1.42 0.11 29.04 5.97
3 350.05 58.50 1.57 0.26 35.19 11.56
Zein 255.60 9.61 1.11 0.03 10.19 2.19
a
Mean standard deviation based on triplicate determinations.
Table 6
Powder owability from the Carrs index and Hausner ratio (Turchiuli et al., 2005).
Carrs index Flowability Hausner ratio
10 Excellent 1.00e1.11
11e15 Good 1.12e1.18
16e20 Fair 1.19e1.25
21e25 Passable 1.26e1.34
26e31 Poor 1.35e1.45
32e37 Very poor 1.46e1.59
>38 Awful >1.60
Table 7
ANOVA and regression coefcients for the bulk density of ax oil encapsulated with
zein after spray drying.
Effect SS p Regression coefcients p
Constant (b
0
) 240.341 7.898 0.021
x
1
8563.80 0.0034** b
1
37.780 10.197 0.168
ns
x
2
1931.06 0.0416* b
2
17.940 10.197 0.329
ns
x
1
2
0.03 0.9915
ns
x
2
2
1.03 0.9488
ns
x
1
x
2
821.37 0.1258
ns
Error 881.69
Total SS 12,198.95
R
2
0.9277 0.8603
Lack of t 0.887
ns
**p 0.01; *p 0.05; ns: Non-signicant; df: Degree of freedom; SS: Sum of
squares.
S. Quispe-Condori et al. / LWT - Food Science and Technology 44 (2011) 1880e1887 1885
ANOVA (Table 7) results showed that the linear effects of zein
and ax oil concentrations on the owing properties were signi-
cant (p < 0.05). The effect of ax oil and zein concentrations is
shown in the contour plot for the bulk density (Fig. 1(c)). The test of
lack of t (p > 0.05) indicates the adequacy of the response model.
It is also observed that the bulk density decreases with an increase
in zein concentration at the same ax oil concentration. The results
obtained for Hausner ratio and Carr index are not presented
because of the low R
2
values and the signicant lack of t of the
response model.
3.4. Microstructures of ax oil microcapsules
SEM images of the zein powder and ax oil microcapsules after
spray and freeze drying are shown in Fig. 3. The surface
morphology of zein powder showed that the particles are irregu-
larly shaped with an internal porous network (Fig. 3(a)). This
observation is similar to that presented by Zhong, Jin, Davidson, and
Zivanovic (2009).
The morphology of ax oil microcapsules depends on the
zein:ax oil ratio and the process used for the microencapsulation.
Flax oil microcapsules prepared by spray drying appeared to be
composed of heterogeneous spheres of various sizes at high
zein:ax oil ratios (Fig. 3(b)). However, at low ratios, a substantial
level of wrinkled surface was observed (Fig. 3(c)). Microcapsules
with wrinkled surfaces were also observed for ax oil xanthan
gum (Omar et al., 2009) and sunower oil milk protein isolate-
dextrin (Ahn, Kim, Lee et al., 2008). Hausner (1967) afrmed that
low Hausner ratio and high compressibility (Carrs Index) were
obtained when the particle deviates from the spherical shape.
On the other hand, microcapsules prepared by freeze drying
resulted in small spheres. Furthermore, the particles appear to be
agglomerated microcapsules (Fig. 3(d)). This behavior was also
observed by Parris et al. (2005) for the microencapsulation of
essential oils with zein by freeze drying. In the microencapsulation
of marine oil with alginate/starch blends, Tan, Chan, and Heng
(2009) observed that microspheres with excessive amounts of
surface oil promoted signicant agglomeration with reduced
microencapsulation efciency. Table 2 and Fig. 3(d) demonstrated
that freeze drying was not efcient due to the high surface oil
content of the particles.
4. Conclusions
Flax oil microcapsules were produced using zein as a coating
material by spray and freeze drying. Using the Central Composite
Design eFace Centered, it was demonstrated that particle yield and
microencapsulationefciency were affected by the zein:axoil ratio
in the spray drying process. Particle yield was signicantly affected
by the zein concentration (linear, p 0.008), while microencapsu-
lation efciency was affected by ax oil concentration (linear,
p 0.013). Furthermore, the statistical model in this study predicted
successfully the microencapsulation efciency. On the other hand,
despite the high particle yield, freeze-dried microcapsules showed
low microencapsulation efciency. Morphology of the microcap-
sules depended onthe zein:axoil ratio and the process used for the
microencapsulation. This study provides optimal formulations to
study other systems using zein as a coating material.
Acknowledgment
The authors are grateful to Alberta Agricultural Research Insti-
tute for nancial support of this project.
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