International Journal of Biological Macromolecules 38 (2006) 25–30

Microencapsulation of ␣-tocopherol using sodium

alginate and its controlled release properties
Sang-Ho Yoo a , Young-Bin Song b , Pahn-Shick Chang c , Hyeon Gyu Lee b,∗
a Department of Food Science and Technology, Sejong University, 98 Gunja-dong, Gwangjin-gu, Seoul 143-747, Republic of Korea
bDepartment of Food and Nutrition, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul 133-791, Republic of Korea
c Department of Food Science and Technology, Seoul National University of Technology, 172 Gongneung-dong, Nowon-gu, Seoul 139-743, Republic of Korea

Received 6 October 2005; received in revised form 8 December 2005; accepted 14 December 2005
Available online 18 January 2006

Abstract
Response surface methodology (RSM) was used to optimize microencapsulation yield (MY) using three independent variables; the ratio of coating
material to core material (w/w, X1 ), the emulsifier concentration (%, v/v, X2 ), and the CaCl2 concentration (%, w/v, X3 ). In the preparation of sodium
alginate (SA) microcapsule, the regression model equation for the MY was predicted as follows; MY(%) = 56.02 + 3.64X2 + 3.18X1 X2 − 3.74X22 .
The optimal conditions for the SA microcapsule were obtained at the [SA]/[␣-TP] ratio of 6.6:3.4 (w/w), [emulsifier] of 1.35% (v/v), and [CaCl2 ]
of 4.3% (w/v), and the predicted MY in this condition was of 57.2%. In vitro ␣-TP releasing test of the SA-based microcapsules was performed.
The SA microcapsule released 28.8% of ␣-TP when exposed in the simulated gastric fluid (SGF, pH 1.2) for 24 h. In the simulated intestinal fluid
(SIF, pH 7.4), the amount of released ␣-TP (81.5%) was significantly greater than that in the SGF. The duration time required for releasing 50 (T50% )
and 70% (T70% ) of ␣-TP from the SA-microcapsule were calculated to be 3.8 and 12.3 h, respectively. From these results, it was suggested that SA
microcapsule would be structurally resistant against acidic environment, and it would rapidly release core material under mild alkali condition.
© 2005 Elsevier B.V. All rights reserved.

Keywords: ␣-Tocopherol (␣-TP); Microencapsulation; Sodium alginate; Response surface methodology (RSM); In vitro releasing test

1. Introduction occur during storage [6]. These impediments to ␣-TP applica-

Among the various Vitamin E categories, ␣-tocopherol (␣-
TP) is a representative antioxidant that dissolves in oil [1].
Alpha-TP has been used as a food additive to provide its antiox-
idant role and also as a functional material to prevent cardio-
vascular diseases and cancer [2,3]. However, the utilization of
beneficial effects of ␣-TP is limited because this compound is
labile to heat and oxygen, then irreversibly converted to quinone
via epoxide formation when is exposed to heat and oxygen [4,5].
In addition, the lipophilic ␣-TP does not dissolve in water and
thus should be assisted by surfactant or emulsifier to increase
bioavailability. Typical dissolving method of lipophilic com-
pounds is to generate an emulsion by homogenizing the mixture
of targeted material and emulsifier in water, but the resulting
vehicle size is generally too large and cracking and creaming
∗ Corresponding author. Tel.: +82 2 2220 1202; fax: +82 2 2292 1226.
E-mail address: hyeonlee@hanyang.ac.kr (H.G. Lee).
tion can be partially overcome by applying microencapsulation
technology to protect ␣-TP from unfavorable environment and
to solubilize it in aqueous environment.
Alginate, one of linear anionic polysaccaharides, consists
of ␤-1,4-d-mannuronic acid (M-block) and ␣-1,4-l-glucuronic
acid (G-block), which is derived from brown seaweeds such as
Laminiaria digitata and L. hyperboria [7]. Sodium alginate is
able to form a gel structure, so called “egg-box model”, when
sodium ion is replaced by a divalent cation like calcium [8].
Alginate gel structure is relatively stable at acidic pH, but it is
easily swollen and disintegrated under mild alkali conditions
[9]. Thus, alginate gel has been applied to produce an effective
controlled release carrier [10–13] as forms of matrix, bead, and
microcapsule [8,14–17].
The microencapsulation techniques, such as spraying drying,
spray chilling, extrusion, coacervation, cocrystallization, and
freeze-drying, have been widely applied in the food industry for
encapsulating vitamins, minerals and other sensitive ingredients
[18,19]. Solvent extraction is another way to produce nutri-

0141-8130/$ – see front matter © 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.ijbiomac.2005.12.013
26 S.-H. Yoo et al. / International Journal of Biological Macromolecules 38 (2006) 25–30

ent delivery system (NDS) but this method uses toxic organic
solvents, causes environmental pollution, and generates high
processing cost. Spray drying seems to be an ideal method for
food application but heat-labile core materials can be severely
damaged during heat-involving manufacturing process [20]. To
accomplish the applicable physical property of coating material,
the ionic gelation and size extrusion method was adopted in our
study. Using this method microcapsules were manufactured by
extruding dispersion mixture of sodium alginate (coating mate-
rial) and ␣-TP (core material) into a hardening agent solution.
Because this whole process proceeded in an aqueous system,
residual solvent does not exist when used in foods. The size
of microcapsules can be controlled by using various sizes of
extruder and changing atomizing pressure [21].
Ultimate goal of our research was to develop an effective
nutrient delivery system (NDS) using a microencapsulation
technique for achieving the controlled release and improving
bioavailability of a model nutrient, ␣-tocopherol (␣-TP). In this
present study, sodium alginate was used as a coating material for
producing microcapsule, and optimal microencapsulation yield
of ␣-TP was obtained. Furthermore, in vitro releasing property of
core material, ␣-TP, was investigated to show applicable poten- Fig. 1. Preparation procedure of sodium alginate (SA) microcapsules.
tial of alginate as an effective nutrient delivery carrier.
capsule was completely destructed by the following procedure:
2. Materials and methods Fifty milligrams of the prepared microcapsule was accurately
weighed and dispersed in 50 mL of ethanol. This dispersion was
2.1. Materials agitated at 700 rpm for 12 h, and then ultrasonic-treated twice for
20 min with 10-min interval. The supernatant after centrifuga-
Sodium alginate (low viscosity) and ␣-tocopherol (from veg- tion at 1800 × g for 10 min was obtained from the disintegrated
etable oil) were purchased from Sigma–Aldrich Chemical Co. microcapsule dispersion, and the amount of ␣-TP was quanti-
(St. Louis, MO, USA). Tween 20 and calcium chloride were sup- fied by measuring UV absorbance at 285 nm. The actual yield of
plied from Wako Chemical Co. (Japan). Other chemicals were microencapsulation was calculated using the equation below:
reagent grade and used without further purification. Actual amount of ␣-TP in the microcapsule
Yield (%) =
Theoretical amount of ␣-TP in the microcapsule
2.2. Microencapsulation of α-tocopherol
×100
The oil in water (O/W) emulsion consisting of sodium algi-
nate (SA) as a coating material, ␣-tocopherol (␣-TP) as a core 2.4. Experimental designs for response surface
material, and Tween 80 as an emulsifier were prepared following methodology (RSM)
the procedure of Lee et al. [22] with some modifications. Basi-
cally, the microcapsules were produced by ionic gelation and The process optimization of ␣-TP encapsulation was
size extrusion technique as shown in Fig. 1. The preparation of designed by central composite design. The response surface
the emulsion was completed by heating the mixture of coating methodology (RSM) was applied to optimize the encapsula-
material and emulsifier in a 45 ◦ C water bath for 1 h, then fol- tion yield of ␣-TP by means of three independent variables;
lowed by constant agitation with an overhead stirrer (Wheaton the ratio of core material to coating material (X1 ), emulsifier
Science Products Co., Millville, USA) at 2000 rpm. The result- concentration (v/v, X2 ), and CaCl2 concentration (w/v, X3 ).
ing O/W emulsion was sprayed into CaCl2 solution by using The actual variable was coded to facilitate multiple regression
an air atomizing system (Spraying system Co., Korea) that was analysis (Table 1). The least-square regression model was ade-
connected to an air compressor (Fig. 2). The prepared microcap- quately fitted into the responses taken from the experimental
sule was hardened at room temperature for 30 min, washed with data and to define an optimization process of the encapsulation
distilled water, sieved with 200 meshes, and then lyophilized. yield.
The optimization conditions were studied at the centre of
2.3. Encapsulation yield of α-TP the design to find the accurate results of statistical experimenta-
tion. At the beginning of the studies, the second-order composite
The encapsulation yield of ␣-TP was determined, following design, with fractional 23 factorial points (eight points), star
the methods of Gohel and Amin [23] and Jee et al. [24] with points (six points) and central point (two points), was effective
some modifications. The coating material structure of micro- in searching for the direction of optimal domain. The regression
 S.-H. Yoo et al. / International Journal of Biological Macromolecules 38 (2006) 25–30 27

Fig. 2. Schematic diagram of air atomizing system for microcapsule preparation.

model equation for the microencapsulation yield of ␣-TP could incubator at 100 rpm and 37 ◦ C. Once the microcapsules were
be predicted as follows: dispersed, 1 mL of the dispersion was taken at designated time
intervals and the same volume of simulated fluid was supple-


3

3

2

3
mented. The ␣-TP released into the simulated fluid was extracted
MY = b0 + bi Xi + bii Xi2 + bij Xi Xj
i=1 i=1 i=1 j=i+1
with 1 mL of chloroform, and the amount of extracted ␣-TP was
determined by UV absorbance at 285 nm. This experiment was
where MY (%) was the microencapsulation yield; subscripts i repeated three times and the average value was calculated.
and j took values from 1 to the number of variables (n); the b0
was the intercept term; the bi values were linear coefficient; the
bij values were quadratic coefficient; xi and xj were the level of 3. Results and discussion
the independent variables. The analysis of data was carried out
using statistical analysis system (SAS) program [25]. Response 3.1. Optimization of α-tocopherol microencapsulation by
surface plots were generated by the StatisticalTM software. response surface methodology

2.5. In vitro releasing property of α-TP
The in vitro ␣-TP releasing property was evaluated using the
microcapsule produced under the maximal yield condition of
microencapsulation [26]. To mimic the digestive tract in human
body, 0.05 M HCl (pH 1.2) with 0.2% NaCl was used as a stimu-
lated gastric fluid (SGF) and 0.05 M phosphate buffer (pH 7.4) as
a simulated intestinal fluid (SIF), but this simulation system did
not contained digestive enzymes [27]. To improve the solubility
of released ␣-TP in the simulated fluids, each testing specimen
contained 0.5% of Tween 80. An aliquot of the microcapsule
(150 mg) was dispersed in 25 mL of the simulated fluid, and the
releasing pattern of ␣-TP was monitored for 24 h in a shaking
Based on the central composite design, the encapsulation
yield of each experimental group (total 16 data points, n = 3)
was determined for the coating material, sodium alginate (SA),
and the obtained data were analyzed by SAS. The encapsu-
lation yield of ␣-tocopherol (␣-TP) in SA microcapsules was
in the range from 31.2 to 58.3% as shown in Table 2. The
regression coefficients calculated for the yield of SA-based
microencapsulation by RSREG analysis were shown in Table 3.
Among the linear, quadratic, and crossproduct forms of inde-
pendent variables, X2 , X1 X2 , and X22 were significant at the
level of p < 0.1. Thus, the regression model equation for the
microencapsulation (MY, %) could be predicted as follow:
MY = 56.02 + 3.64X2 + 3.18X1 X2 − 3.74X22 . According to

Table 1
Coded levels for independent variables used in experimental design for microencapsulation of ␣-tocopherol
Variables Coded, Xi Coded level Xa

−2 −1 0 1 2

Ratio of [AG] to [␣-TP]b X1 4:6 5:5 6:4 7:3 8:2 1:−1

Emulsifier concentration (%, w/v) X2 0 0.5 1.0 1.5 2.0 0.5
CaCl2 concentration (%, w/v) X3 3.0 3.5 4.0 4.5 5.0 0.5
a X is the increment of the experiment factor values corresponding to one unit of the coded variable.
b [SA] and [␣-TP] were sodium alginate and ␣-tocopherol, respectively. Sodium alginate concentration used in this study was in the range of 2.0–4.0% (w/v).
28 S.-H. Yoo et al. / International Journal of Biological Macromolecules 38 (2006) 25–30

Table 2
Central composite design for the optimization of ␣-tocopherol microencapsulation prepared with sodium alginate

Run number Coded variablesa Process variablesa Experimental pointb (%)

X1 X2 X3 X1 X2 X3

1 −1 −1 −1 5:5 0.5 3.5 53.86

2 −1 −1 1 5:5 0.5 4.5 54.25
3 −1 1 −1 5:5 1.5 3.5 51.94
4 −1 1 1 5:5 1.5 4.5 53.43
5 1 −1 −1 7:3 0.5 3.5 39.57
6 1 −1 1 7:3 0.5 4.5 43.81
7 1 1 −1 7:3 1.5 3.5 51.24
8 1 1 1 7:3 1.5 4.5 54.81
9 0 0 0 6:4 1.0 4 53.07
10 0 0 0 6:4 1.0 4 58.32
11 −2 0 0 4:6 1.0 4 53.99
12 2 0 0 8:2 1.0 4 50.99
13 0 −2 0 6:4 0 4 31.15
14 0 2 0 6:4 2.0 4 50.33
15 0 0 −2 6:4 1.0 3 46.98
16 0 0 2 6:4 1.0 5 52.84
a The X1 , X2 , and X3 were the ratio of core material to coating material, the emulsifier concentration (%, v/v) and CaCl2 concentration in dispersion fluid (%, w/v),
respectively.
b The experimental point (%) is equivalent to the encapsulation yield of ␣-tocopherol.

Table 3
Values of regression coefficients calculated for the microencapsulation of ␣-tocopherol using sodium alginate as a coating material
Independent variablea Coefficient Standard error t-value Significance level (p)

Constant 56.022500 2.051848 27.30 <0.0001

Linear (r2 = 0.4321** )
X1 −1.879125 0.775526 −2.42 0.0517
X2 3.643125 0.775526 4.70 0.0033**
X3 1.338125 0.775526 1.73 0.1352
Quadratic (r2 = 0.3604* )
X12 −0.81250 0.775526 −1.03 0.3414
X22 −3.738750 0.775526 −4.82 0.0029**
X23 −1.446250 0.775526 −1.86 0.1115
Interaction (r2 = 0.1237)
X1 X2 3.176250 1.096759 2.90 0.0275*
X1 X3 0.741250 1.096759 0.68 0.5243
X2 X3 0.053750 1.096759 0.05 0.9625
r2 0.9161*
F 7.28
Probability of F 0.0126
a The X , X , and X were the ratio of core material to coating material (w/w), the concentration of emulsifier (%) and the concentration of CaCl in the dispersion
1 2 3 2
fluid, respectively.
* p < 0.1.
** p < 0.01.

the model equation, the [emulsifier], X2 , were believed to be
the most important factor affecting the MY among the variables
studied, and the yield was somewhat affected by the ratio of core
material [C1 m] to coating material [C2 m], X1 . The [CaCl2 ] was
shown to have little effect on the microencapsulation yield.
3.2. Response surface plotting and canonical analysis of
microencapsulation yield
The model equation was applied to obtain optimal condi-
tions of ␣-TP encapsulation by the three-dimensional graphical
methodology. The resulting three-dimensional surface plots of
encapsulation yield versus three pairs of two process parameters
(Xi s) were obtained from the SA-based encapsulation (Fig. 3)
and the optimal conditions were determined by canonical analy-
sis. In this analytical process, the coded level of third parameter
not used for the plotting was placed at zero level to interpret the
effect of two independent variables on the MY. As the eigen-
values were all negative in the canonical analysis, the stationary
point was the maximal one. Consequently, the critical values
reflected optimal conditions for the microencapsulation, and
each independent variable for the maximal MY (%) of ␣-TP
 S.-H. Yoo et al. / International Journal of Biological Macromolecules 38 (2006) 25–30 29

Fig. 4. The release profiles of ␣-TP from SA microcapsule in simulated gastric

fluid (SGF) and simulated intestinal fluid (SIF).

was selected as shown in Table 4. Applying the selected inde-

pendent variables, 57.2% of MY was expected as maximal yield.
This predicted value was close to 56.7% of MY, as counted from
the actual experimental observations, which was somewhat less
than predicted.

3.3. In vitro α-TP releasing property of microcapsule

The desirable microcapsule for the nutrient delivery sys-

tem (NDS) should protect core material without acid-damaged
destruction under strong acidic environment, which means that
the coating material should be resistant against acid. Once the
microcapsule reach small intestine, it should initially release the
core material rapidly, and keep the releasing rate to maintain the
effective concentration of core material as the absorption rate
decreases [28].
The SA-based microcapsules produced under the optimal
yield conditions were applied for in vitro ␣-TP releasing experi-
ment in the SGF (simulated gastric fluid, pH 1.2) and SIF (simu-
lated intestinal fluid, pH 7.4) for 24-h period. The 28.8% of ␣-TP
was released from SA-based macrocapsules in the SGF during
24 h (Fig. 4). In the SIF, 81.5% of ␣-TP was finally released and
more than 50% of ␣-TP was released within 4 h. More specif-
ically, the duration time required for releasing 50 (T50% ) and
70% (T70% ) of ␣-TP from the SA-microcapsule were predicted
to be 3.8 and 12.3 h, respectively, from the releasing profile of
␣-TP. Thus, it was clear that SA-based microcapsules were quite
stable under acidic condition, but it was easily swollen and disin-

Table 4
Critical values from canonical analysis of response surface based on the coded
data for the SA microcapsule
Fig. 3. (A) Response surfaces for the effect of the [SA]/[␣-TP] ratio (X1 ) and the
concentration of emulsifier (X2 ) on the yield of microencapsulation. The [SA] Variablea Coded factor value Uncoded factor value
and [␣-TP] represent the concentrations of sodium alginate and ␣-tocopherol,
X1 0.5798 6.6:3.4
respectively. (B) Response surfaces for the effect of the [SA]/[␣-TP] ratio
X2 0.7380 1.35
(X1 ) and the concentration of CaCl2 in dispersion fluid (X3 ) on the yield of
X3 0.6249 4.3
microencapsulation. (C) Response surfaces for the effect of the concentration
of emulsifier (X2 ) and the concentration of CaCl2 in dispersion fluid (X3 ) on the a X , X and X were the ratio of coating material to core material (w/w),
1 2 3
yield of microencapsulation. the emulsifier concentration (%, v/v) and the CaCl2 concentration in dispersion
fluid (%, w/v), respectively.
30 S.-H. Yoo et al. / International Journal of Biological Macromolecules 38 (2006) 25–30
tegrated under the neutral or near alkaline condition [9]. It was
previously reported that from chitosan-coated alginate beads,
less than 10% of Sudan orange G in the SGF and around 85% of
it was released in the SIF [26]. Therefore, it was concluded that
the sodium alginate as a coating material was structurally resis-
tant against acidic environment, and rapidly released targeted
core material, ␣-TP, under mild alkali condition.
Acknowledgement
This work was supported by the Korea Research Foundation
Grant (KRF-2003-041-C00415).
References
[1] K.U. Ingold, A.C. Webb, D. Witter, G.W. Burton, T.A. Metcalfe, D.P.R.
Muller, Arch. Biochem. Biophys. 259 (1987) 224.
[2] W.J. Mergens, J. Chau, H.L. Newmark, IARC Sci. Publ. 31 (1980) 259.
[3] K.F. Gey, Int. J. Vit. Nutr. Res. Suppl. 30 (1989) 224.
[4] T. Verleyen, R. Verhe, A. Huyghebaert, K. Dewettinck, W.D. Greyt, J.
Agric. Food. Chem. 49 (2001) 1508.
[5] D. Barrera-Arellano, V. Ruiz-Mendez, G. Ruiz, C. Dobarganes, J. Sci.
Food. Agric. 79 (1999) 1923.
[6] T. Julianto, K.H. Yuen, A.M. Noor, Int. J. Pharm. 200 (2000) 53.
[7] A. Kikuchi, M. Kawabuchi, M. Sugihara, Y. Sakurai, J. Control. Release
47 (1997) 21.
[8] N. Segi, T. Yotsuyanagi, K. Ikeda, Chem. Pharm. Bull. 37 (1989) 3092.
[9] F. Acarturk, S. Takka, J. Microencapsul. 16 (1999) 291.
[10] C.K. Kim, E.J. Lee, Int. J. Pharm. 79 (1992) 11.
[11] H. Tomida, C. Nakamura, H. Yoshitomi, S. Kiryu, Chem. Pharm. Bull.
41 (1993) 1261.
[12] S. Sugawara, T. Imai, M. Otagiri, Pharm. Res. 11 (1994) 272.
[13] B.J. Lee, G.H. Min, Int. J. Pharm. 144 (1996) 37.
[14] M. Johnson, J. Medlin, Eur. Polym. J. 21 (1985) 147.
[15] M. Nakano, A. Ogata, Chem. Pharm. Bull. 32 (1984) 782.
[16] A.F. Stockwell, S.S. Davis, S.E. Walker, J. Control. Release 3 (1986)
167.
[17] T. Yotsuyanagi, T. Ohkubo, T. Ohhashi, K. Ikeda, Chem. Pharm. Bull.
35 (1987) 1555.
[18] L.L. Balassa, G.O. Fanger, Crit. Rev. Food Technol. 2 (1971) 245.
[19] J.D. Dziezak, Food Technol. 42 (1988) 136.
[20] E.M. Hong, M.G. Yu, B.S. Noh, P.S. Chang, Kor. J. Food Sci. Technol.
34 (2002) 437.
[21] J.K. Vasir, K. Tambwekar, S. Garg, Int. J. Pharm. 255 (2003) 13.
[22] D.W. Lee, S.J. Hwang, J.B. Park, H.J. Park, J. Microencapsul. 20 (2003)
179.
[23] M.C. Gohel, A.F. Amin, J. Control. Release 51 (1998) 115.
[24] U.K. Jee, U.J. Joen, G.W. Lee, K. Han, Y.B. Chung, J. Kor. Pharm. Sci.
24 (1994) 167.
[25] SAS Institute, Statistical Analysis System, Version 6.12, SAS Institute
Inc., Cary, NC, 1990.
[26] A.J. Ribeiro, R.J. Neufeld, P. Arnaud, J.C. Chaumeil, Int. J. Pharm. 187
(1999) 115.
[27] D. Wong, S. Larrabee, K. Clifford, J. Tremblay, D.R. Friend, J. Control.
Release 47 (1997) 173.
[28] J.Y. Kang, K.T. Lee, S.H. Seo, J. Kor. Pharm. Sci. 28 (1998) 109.