International Journal of Biological Macromolecules 51 (2012) 10701078

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International Journal of Biological Macromolecules

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Calcium alginate/gum Arabic beads containing glibenclamide: Development and

in vitro characterization
Amit Kumar Nayak, Biswarup Das , Ruma Maji
Department of Pharmaceutics, Seemanta Institute of Pharmaceutical Sciences, Mayurbhanj 757086, Odisha, India

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

Article history: This work investigates the development, optimization and in vitro characterization of calcium
Received 29 June 2012 alginate/gum Arabic beads by an ionotropic gelation method for prolonged sustained release of gliben-
Received in revised form 29 July 2012 clamide. The effects of amount of sodium alginate and gum Arabic as independent process variables on the
Accepted 19 August 2012
drug encapsulation efciency and drug release were optimized and analyzed based on central composite
Available online 27 August 2012
design and response surface methodology. Increment in drug encapsulation efciency and decrease in
drug release were found with the increase of both the amounts of sodium alginate and gum Arabic, used
Keywords:
as polymer-blend. These optimized beads showed high drug encapsulation efciency (86.02 2.97%),
Calcium alginate
Gum Arabic
and suitable sustained drug release pattern over prolonged period (cumulative drug release after 7 h
Sustained release of 35.68 1.38%). The average size of these formulated dried beads containing glibenclamide ranged
Optimization from 1.15 0.11 to 1.55 0.19 mm. The in vitro dissolution of these beads showed prolonged sustained
Central composite design release of glibenclamide over 7 h, which followed rst-order model (R2 = 0.98860.9985) with anoma-
Response surface methodology lous (non-Fickian) diffusion mechanism (release exponent, n = 0.720.81). The swelling and degradation
of the optimized beads were inuenced by pH of test mediums. These beads were also characterized
by SEM and FTIR spectroscopy for surface morphology and excipients-drug interaction analysis, respec-
tively. These developed calcium alginate/gum Arabic beads containing glibenclamide could possibly be
advantageous in terms of advanced patient compliance with reduced dosing interval.
2012 Elsevier B.V. All rights reserved.

1. Introduction through ionotropic gelation [3,4]. Alginate undergo ionotropic gela-

Currently, the exploration of natural polymers in the devel-
opment of various pharmaceutical dosage forms has advanced
consistently in terms of diversity and property. The natural poly-
mers offer certain specic advantages over synthetic polymers,
such as easy availability, biocompatibility, biodegradability, non-
toxicity and pollution free processing [1,2]. Over the past few
years, a great deal of attention has been paid in the development
of biopolymeric beads for use as drug delivery carriers through
ionotropic gelation technique [3,4]. The ionic polymers (alginate,
pectin, gellan gum, etc.) undergo ionotropic gelation and pre-
cipitate to form beads due to electrostatic interaction between
oppositely charged species. This technique is very simple and the
conditions used were very mild. In addition, physical cross-linking
due to ionotropic-gelation instead of chemical cross-linking avoids
the possible toxicity of reagents and other undesirable effects [3].
Among various ionic biopolymers, alginate, a polyanionic
copolymer of mannuronic and guluronic acid residues has been
investigated for its unique nature of forming hydrogel beads
Corresponding author. Tel.: +91 9583131603.
E-mail address: biswarup777@gmail.com (B. Das).
tion in the presence of divalent cations like Ca2+ , Ba2+ , Cu2+ , Pb2+ ,
Zn2+ , Cd2+ , etc and trivalent cations like Al3+ , etc due to ionic
interaction between carboxylic acid groups of alginate and these
cations [5,6]. Various drugs have been successfully encapsulated
in ionotropically gelled alginate beads and exhibited different drug
release proles [7,8]. Although, ionotropically gelled alginate beads
can be prepared by simple and mild procedures, this method has a
major limitation of drug loss during bead preparation due to leach-
ing of drugs through the pores [9]. Therefore, many modications
of ionotropically gelled alginate beads based on the use of another
polymer as blend with alginate were investigated for drug delivery
applications [2,1018].
Gum Arabic, a biocompatible and biodegradable natural gum,
is mainly used in oral and topical pharmaceutical formulations
[19]. The popularity of gum Arabic is due to physical properties
including high solubility, pH stability, non-toxicity and gelling
characteristics [20]. According to United States Food and Drug
Administration (USFDA), gum Arabica enjoys the Generally Rec-
ognized as Safe (GRAS) status [20]. It is highly branched and
slightly acidic polysaccharide found as a mixed calcium, magne-
sium, and potassium salt of polysaccharidic acids with main chain
of (13)--d-galactopyranosyl units and side chains containing
l-arabinofuranosyl, l-rhamnopyranosyl, d-galactopyranosyl, and

0141-8130/$ see front matter 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.ijbiomac.2012.08.021
 A.K. Nayak et al. / International Journal of Biological Macromolecules 51 (2012) 10701078 1071

d-glucopyranosyl uronic acid units [20,21]. In the current inves- Table 1

Factors and levels of the central composite design used for optimization.
tigation, the combination of sodium alginate and another natural
polymer, gum Arabic was used as polymer-blend to develop Normalized levels Experimental settings
ionotropically gelled calcium alginate/gum Arabic beads for use
Sodium alginate (mg) Gum Arabic (mg)
in sustained drug release application. However, any report on (A) (B)
alginate-gum Arabic blend beads is unavailable in the previous lit-
1.414 179.29 79.29
erature, which adds the work novel of its kind. Glibenclamide was 1 200.00 100.00
used as a model drug in the present study to evaluate the sustained 0 250.00 150.00
drug release potential of the calcium alginate/gum Arabic beads 1 300.00 200.00
prepared using ionotropic gelation. 1.414 320.71 220.71

Glibenclamide, a second generation sulfonylurea, is used in the

treatment of non-insulin dependent diabetes mellitus (Type-II)
[22]. Its plasma half-life is 46 h, which makes multiple dosing to
maintain the therapeutic blood level [23]. To reduce multiple dos-
ing, sustained release dosage of glibenclamide, which will be able
to deliver glibenclamide at a slow release rate over an extended
period is essential.
Optimization by means of statistical experimental design
methodologies has been widely applied by designing a set of exper-
iments that will reliably measure the response variables, tting a
mathematical model to the data, conducting appropriate statistical
test to assure that the best possible model is chosen, and deter-
mining the values of independent formulation variables to produce
optimum response [24]. Among various statistical optimization
designs, central composite design, which is a response surface
design, has been widely used for formulation and process opti-
mization [13,16]. It is very efcient and exible, providing much
information on experiment variable effects and overall experi-
mental error in a minimal number of required runs [25]. In the
current investigation, the central composite design was employed
for the formulation optimization of ionotropically gelled calcium
alginate/gum Arabic beads containing glibenclamide.
2. Experimental
2.1. Materials
Glibenclamide (B.S. Trader Pvt. Ltd., India), sodium alginate
(Central Drug House, India), gum Arabic (Central Drug House,
India), and calcium chloride (Mark Specialties Pvt. Ltd., India) were
used. All chemicals and reagents used were of analytical grade.
2.2. Preparation of calcium alginate/gum Arabic beads containing
glibenclamide
prepared separately using distilled water. These dispersions were
well mixed with stirring for 10 min at 1000 rpm using a magnetic
stirrer (Remi Motors, India). Afterwards, glibenclamide was added
to the dispersion mixture. The ratio of drug to polymer was
maintained 1:4 in all formulations and mixed thoroughly using a
homogenizer (Remi Motors, India). The nal sodium alginate/gum
Arabic gels containing glibenclamide were ultrasonicated for 5 min
for debubbling. The resulting dispersion was then added via a
21-gauge needle drop wise into100 ml of 10% (w/v) CaCl2 solution.
Added droplets were retained in the CaCl2 solution for 15 min
to complete the curing reaction and to produce rigid beads. The
wet beads were collected by decantation, and washed two times
with distilled water and dried at 37 C for overnight. The calcium
alginate/gum Arabic beads containing glibenclamide were stored
in a desiccator until used.
2.3. Experimental design
A central composite design (spherical type, single center point,
and = 1.414) was employed for the formulation optimization of
calcium alginate/gum Arabic beads containing glibenclamide. The
amount of sodium alginate (A) and the gum Arabic (B) as poly-
meric blend were dened as the selected independent formulation
variables (factors); while drug encapsulation efciency (DEE, %),
and drug release at 7 h (R7 h , %) were used as dependent variables
(responses). The process variables (factors) and levels with exper-
imental values are reported in Table 1. The matrix of the design
including investigated factors and responses are also shown in
Table 2. Design-Expert 8.0.6.1 software (Stat-Ease Inc., USA) was
used for generation and evaluation of experimental design. The
polynomial mathematical model generated by circumscribed cen-
tral composite design is following [25]:
Y = b0 + b1 A + b2 B + b3 AB + b4 A2 + b5 B2 ,

The calcium alginate/gum Arabic beads containing gliben- where Y is the response; b0 is the intercept, and b1 , b2 , b3 , b4 , b5
clamide were prepared using ionotropic gelation method. Calcium are regression coefcients. A and B are individual effects; A2 and B2
chloride (CaCl2 ) was used as cross-linker in ionotropic gelation. are quadratic effects; AB is the interaction effect. One-way ANOVA
Briey, sodium alginate and gum Arabic aqueous dispersions were was applied to estimate the signicance of the model (p < 0.05). The

Table 2
Experimental plan and observed response values from randomized run in central composite design.

Experimental formulations Factors Responsesa

Sodium alginate (mg)(A) Gum Arabic (mg)(B) DEE (%)b R7 h (%)c

F-1 200.00 100.00 54.17 1.54 53.23 2.22

F-2 200.00 200.00 67.36 2.15 44.24 2.03
F-3 300.00 100.00 60.25 2.12 49.76 2.16
F-4 300.00 200.00 70.28 3.04 43.75 1.77
F-5 179.29 150.00 59.13 2.22 50.81 1.97
F-6 320.71 150.00 65.80 1.98 46.36 2.03
F-7 250.00 220.71 73.93 3.37 40.54 1.54
F-8 250.00 79.29 55.38 1.76 53.06 1.98
F-9 250.00 150.00 58.04 2.05 51.27 2.07
a
Observed response values: Mean S.D. (n = 3).
b
DEE (%) = Drug encapsulation efciency (%).
c
R7h (%) = drug release at 7 h.
1072 A.K. Nayak et al. / International Journal of Biological Macromolecules 51 (2012) 10701078
surface response plots and contour plots were analyzed to reveal
the effect of independent factors (amount of sodium alginate and
gum Arabic ratio) on the measured responses (DEE, and R7 h ) were
analyzed.
2.4. Determination of DEE (%)
Accurately weighed 100 mg of prepared beads from each batch
were taken separately and were crushed using pestle and mortar.
The crushed powders were placed in 500 ml of phosphate buffer, pH
7.4, and kept for 24 h with occasionally shaking at 37 0.5 C. After
the stipulated time, the mixture was stirred at 500 rpm for 20 min
using a magnetic stirrer. The polymer debris formed after the dis-
integration of beads was removed by ltering through Whatman
lter paper (No. 40). The drug content in the ltrate was determined
using a UVvis spectrophotometer (Shimadzu, Japan) by measur-
ing absorbance at Max of 228 nm. The DEE of beads was calculated
using the following formula:
actual drug content in beads
DEE(%) = 100.
theoretical drug content in beads
2.5. Determination of bead size
Particle size of 100 dried beads from each batch was measured
by optical microscopic method for average particle size using an
optical microscope (Olympus). The ocular micrometer was previ-
ously calibrated by stage micrometer.
2.6. Surface morphology analysis
The surface morphology of the formulated beads was analyzed
by scanning electron microscope (SEM) (JEOL, JSM-5800, Japan).
Beads were gold coated by mounted on a brass stub using double-
sided adhesive tape and under vacuum in an ion sputter with a thin
layer of gold (35 nm) for 75 s and at 15 kV to make them electrically
conductive and their morphology was examined.
2.7. Fourier transform-infrared (FTIR) spectroscopy
Samples were reduced to powder and analyzed as KBr pellets
by using a Fourier transform-infrared (FTIR) spectroscope (Perkin
Elmer Spectrum RX I, USA). The pellet was placed in the sam-
ple holder. Spectral scanning was taken in the wavelength region
between 3800 and 400 cm1 at a resolution of 4 cm1 with scan
speed of 1 cm/s.
2.8. In vitro drug release studies
The release of the glibenclamide from various ionotropically
gelled calcium alginate/gum Arabic beads containing gliben-
clamide was tested using a dissolution apparatus USP/BP/IP
(Campbell Electronics, India). The baskets were covered with 100-
mesh nylon cloth to prevent the escape of the beads. The dissolution
rates were measured at 37 1 C under 50 rpm speed. Accurately
weighed quantities of calcium alginate/gum Arabic beads contain-
ing glibenclamide equivalent to 5 mg glibenclamide were added
to 900 ml of simulated gastric uid (pH 1.2). The test was carried
out in simulated gastric uid (pH 1.2) for 2 h, and then, continued
in simulated intestinal uid (pH 7.4) for next 5 h. 5 ml of aliquots
was collected at regular time intervals, and the same amount of
fresh dissolution medium was replaced into dissolution vessel to
maintain the sink condition throughout the experiment. The col-
lected aliquots were ltered, and suitably diluted to determine the
absorbance using a UVvis spectrophotometer (Shimadzu, Japan)
by measuring absorbance at Max of 228 nm.
2.9. Analysis of in vitro drug release kinetics and mechanism
In order to predict and correlate the in vitro drug release behav-
ior from formulated calcium alginate/gum Arabic beads containing
glibenclamide, it is necessary to t into a suitable mathematical
model. The in vitro drug release data were evaluated kinetically
using various important mathematical models like zero order,
rst order, HixsonCrowell, Weibull, BakerLonsdale, Higuchi, and
KorsmeyerPeppas models [26].
Zero-order model: Q = kt + Q0 ; where Q represents the drug
released amount in time t, Q0 is the start value of Q, and k is the
rate constant.
First-order model: Q = Q0 ekt ; where Q represents the drug released
amount in time t, Q0 is the start value of Q, and k is the rate constant.
1/3
HixsonCrowell model: Q 1/3 = kt + Q0 ; where Q represents the
drug-released amount in time t, Q0 is the start value of Q, and k is
the rate constant.
Weibull model: m = 1 exp [(t)b/a ]; where m represents the drug
released amount in time t, a is the time constant, and b is the shape
parameter.
BakerLonsdale model: 3/2 [1 (1 Q)2/3 ] Q = kt; where Q repre-
sents the drug released amount in time t, and k is the rate constant.
Higuchi model: Q = kt0.5 ; where Q represents the drug released
amount in time t, and k is the rate constant.
KorsmeyerPeppas model: Q = ktn ; where Q represents the drug
released amount in time t, k is the rate constant, and n is the
diffusional exponent, indicative of drug release mechanism.
The accuracy and prediction ability of these models were com-
pared by calculation of squared correlation coefcient (R2 ) using
KinetDS 3.0 Rev. 2010 software.
Again, The KorsmeyerPeppas model was employed in the in
vitro drug release behavior analysis of these formulations to dis-
tinguish between competing release mechanisms: Fickian release
(diffusion-controlled release), non-Fickian release (anomalous
transport) and case-II transport (relaxation-controlled release).
When n is 0.43, it is Fickian release. The n value between 0.43 and
0.85 is dened as non-Fickian release. When, n 0.85, it is case-II
transport [13,15,16].
2.10. Swelling behavior measurement
Swelling measurement of optimized calcium alginate/gum Ara-
bic beads containing glibenclamide was carried out in two different
aqueous media: simulated gastric uid (pH 1.2), and simulated
intestinal uid (pH 7.4). 100 mg beads were placed in vessels of dis-
solution apparatus (Campbell Electronics, India) containing 500 ml
respective media. The experiment was carried out at 37 1 C
under 50 rpm paddle speed. The swelled beads were removed at
predetermined time interval and weighed after drying the surface
by using tissue paper. Swelling index was determined using the
following formula:
Swelling index
weight of beads after swelling dry weight of beads
= 100.
dry weight of beads
 A.K. Nayak et al. / International Journal of Biological Macromolecules 51 (2012) 10701078 1073

Table 3
Summary of ANOVA for the response parameters.

Source Sum of squares d.f.a Mean square F-Value p-Value

Prob > F

(a) For DEE (%)b

Model 379.95 5 75.99 130.06 0.0011 (S)
A 42.47 1 42.47 72.69 0.0034 (S)
B 305.71 1 305.71 523.25 0.0002 (S)
AB 2.50 1 2.50 4.27 0.1306 (NS)
A2 12.54 1 12.54 21.46 0.0189 (S)
B2 29.26 1 29.26 50.08 0.0058 (S)

(b) For R7 h (%)c

Model 163.47 5 32.69 61.25 0.0032 (S)
A 13.14 1 13.14 24.62 0.0157 (S)
B 133.71 1 133.71 250.50 0.0005 (S)
AB 2.22 1 2.22 4.16 0.1341 (S)
A2 5.14 1 5.14 9.63 0.0532 (NS)
B2 14.36 1 14.36 26.91 0.0139 (S)

A and B represent the main effects (factors); A2 and B2 are the quadratic effect; AB is the interaction effect.
a
d.f. = Degree of freedom.
b
DEE (%) = Drug encapsulation efciency (%).
c
R7h (%) = drug release at 7 h.

2.11. Statistical analysis 3.2. Formulation optimization by central composite design

Statistical optimization was performed using Design-Expert Designing pharmaceutical formulations with the minimum
8.0.6.1 software (Stat-Ease Inc., USA). The squared correlation number of trials is very crucial for pharmaceutical scientists [30].
coefcients (R2 ) of all kinetic models were determined using In conventional optimization process by changing one factor at a
KinetDS 3.0 Rev. 2010 software. All measured data are expressed time, the optimization is usually carried out by varying a single fac-
as mean standard deviation (S.D.). Each measurement was done tor and keeping all other factors xed at a specic set of conditions.
in triplicate (n = 3). This method is time consuming and incapable of effective optimiza-
tion as it does not consider the interactive effects of all the primary
factors [31]. It is therefore important to understand the inuence
of formulation variables on the formulation quality with a minimal
3. Results and discussion
number of experimental trials and subsequent selection of formu-
lation variables to develop optimized formulation using established
3.1. Preparation of calcium alginate/gum Arabic beads containing
statistical tools such as central composite design. Central compos-
glibenclamide
ite design is a response surface design, which provides information
on individual effects, pair-wise interactions of various individual
The divalent calcium ions t into electronegative cavities of
effects and curvilinear variables effects [25]. In the current inves-
the sodium alginate like eggs in an Egg Box model to form
tigation, a central composite design (spherical type, single center
calcium alginate due to electrostatic ionic interaction between neg-
point, and = 1.414) with total 9 experimental formulations of cal-
atively charged carboxylate group of sodium alginate and positively
cium alginate/gum Arabic beads containing glibenclamide were
charged calcium ions present in the cross-linking solution [27]. At
proposed by Design-Expert 8.0.6.1 software (Stat-Ease Inc., USA)
the cross-linking sites, polyvalent cations cause interpolysaccha-
for two independent process variables (factors): amount of sodium
ride binding, which are called as junction zones [28]. The calcium
alginate (A) and amount of gum Arabic (B) in the polymer-blend
ions compete with the sodium ions of sodium alginate and thus,
used (Table 1). The effects of these independent variables on DEE
bring the two polymer chains together. Calcium ions are accommo-
(%), and R7 h (%) were investigated as optimization response param-
dated in the interstices of two polyuronate chains having a close
eters. According to the trial proposal of central composite design, 9
ion-pair interaction with carboxylate anions of the sodium algi-
experimental formulations of calcium alginate/gum Arabic beads
nate and sufcient coordination by other electronegative oxygen
containing glibenclamide were prepared using ionotropic gela-
atoms [29]. Besides anionic nature of sodium alginate, gum Arabic
tion method. Overview of the experimental plan and the observed
possesses slightly anionic nature with a mixed calcium, magne-
response values are presented in Table 2. The Design-Expert 8.0.6.1
sium, and potassium salt of polysaccharidic acids with main chain
software provided suitable polynomial model equations involving
of (13)--d-galactopyranosyl units and side chains containing
individual main factors and interaction factors for each investigated
l-arabinofuranosyl, l-rhamnopyranosyl, d-galactopyranosyl, and
responses after tting these data. These models were evaluated sta-
d-glucopyranosyl uronic acid units [20,21]. Therefore, calcium
tistically by applying one-way ANOVA (p < 0.05), which is shown in
cations present in the cross-linking solution might compete with
Table 3. The model p values of less than 0.05 for both the measured
magnesium and potassium cations present in the polysaccha-
responses implied the models were signicant (p < 0.05).
ridic structure of gum Arabic. When these polysaccharides were
The model equation relating DEE (%) as response became:
exposed to divalent calcium cations in the current investigation,
there could occur an electrostatic ionic interaction between posi- DEE(%) = 96.57 0.32 A 0.15 B 3.16 104 AB + 8.31
tively charged calcium cations and negatively charged carboxylate
104 A2 + 1.27 104 B2 [R2 = 0.9954; p < 0.05]
groups of sodium alginate and gum Arabic. Thus, various calcium
alginate/gum Arabic beads containing glibenclamide was prepared The model equation relating R7 h (%) as response became:
through ionotropic gelation method when various dispersions of
R7 h (%) = 27.89 + 0.20 A + 0.11 B + 2.98 104 AB 5.32
sodium alginate, gum Arabic, and glibenclamide were dropped into
the solutions containing calcium ions. 104 A2 8.89 104 B2 [R2 = 0.9903; p < 0.05]
1074 A.K. Nayak et al. / International Journal of Biological Macromolecules 51 (2012) 10701078

Fig. 1. Three-dimensional response surface plots showing the effects of amounts of sodium alginate (mg) and gum Arabic (mg) on (a) DEE (%) and (b) R7 h (%). Two-dimensional
corresponding contour plots showing the effects of amounts of sodium alginate (mg) and gum Arabic (mg) on (c) DEE (%) and (d) R6 h (%).

of gum Arabic (B) in the formulated calcium alginate/gum Arabic

Model simplication was carried out by eliminating non-
signicant terms (p > 0.05) in model equations resulting from the
multiple regression analysis [32], giving:
DEE(%) = 96.57 0.32 A 0.18 B + 8.31 104 A2 + 1.27 104 B2
R7 h (%) = 27.89 + 0.20 A + 0.11 B 8.89 104 B2
Design-Expert 8.0.6.1 software generated three-dimensional
response surface plots and corresponding contour plots relating
investigated responses, DEE (%) and R7 h (%). The three-dimensional
response surface plot is very useful in learning about the main and
interaction effects of the independent variables (factors), whereas
two-dimensional contour graph gives a visual representation of
values of the response [15,33]. The three-dimensional response sur-
face plots relating DEE (%) and R7 h (%) are presented in Fig. 1(a and
b, respectively). The two-dimensional corresponding contour plots
relating DEE (%) and R6 h (%) are presented in Fig. 1(c and d, respec-
tively). The three-dimensional response surface plot relating DEE
(%) (Fig. 1c) depicted the increase in DEE with the increase of both
the amount of sodium alginate (A), and amount of gum Arabic (B).
On the other hand, the three-dimensional response surface plots
relating R7 h (%) (Fig. 1d) also indicated the decrease in R7 h with the
increase of both the amount of sodium alginate (A), and amount
beads containing glibenclamide.
A numerical optimization technique using the desirability
approach was employed to develop new formulations with desired
response (desired quality). A constraint to maximizing the DEE and
minimizing the R7 h was to set the goal to locate the optimum sett-
ings of independent variables for the optimized formula by the
central composite design using the the Design-Expert 8.0.3 soft-
ware based on the criterion of desirability. To get the desired opti-
mum responses, independable variables (factors) were restricted
to 200.00 mg A 400.00 mg, and 100.00 mg B 200.00 mg;
whereas the desirable ranges of responses were restricted to
85% DEE 100%, and 30% R7 h 40%. The desirability plot indi-
cating desirable regression ranges for optimal process variable
settings and overlay plot indicating the region of optimal pro-
cess variable settings are presented in Fig. 2(a and b, respectively).
In order to evaluate the optimization capability of these models
generated according to the results of central composite design,
optimized calcium alginate/gum Arabic beads containing gliben-
clamide were prepared using one of the optimal process variable
settings proposed by the design (R2 = 1). The selected optimal
process variable setting used for the formulation of optimized
formulation was A = 395.13 mg and B = 178.98 mg. The optimized
beads containing glibenclamide (F-O) were evaluated for DEE
(%) and R7 h (%). Table 4 lists the results of experiments with
 A.K. Nayak et al. / International Journal of Biological Macromolecules 51 (2012) 10701078 1075

Fig. 2. The desirability plot: (a) indicating desirable regression ranges and the overlay plot and (b) indicating the region of optimal process variable settings.

Table 4
Results of experiments for conrming optimization capability.

Code Factors Responsesa

Sodium alginate (mg) Gum Arabic (mg) DEE (%)b R7 h (%)c

d
Predicted value Actual value Error (%) Predicted value Actual value Error (%)d

F-O 395.13 178.98 85.24 86.02 2.97 0.92 34.66 35.68 1.38 2.86
a
Observed response values: mean S.D. (n = 3).
b
DEE (%) = Drug encapsulation efciency (%).
c
R7h (%) = drug release at 7 h.
d
Error (%) = [difference between predicted value and actual value/Predicted value] 100.

predicted responses by the mathematical models and those actu- Arabic) solution with incorporation of both the polymers in increas-
ally observed. The optimized beads containing glibenclamide (F-O) ing ratio that in turn increased the droplet size during addition of
showed DEE of 86.02 2.97% and R7 h of 35.68 1.38% with small the polymer blend solution to the cross-linking solution.
error-values (0.92 and 2.86, respectively). This reveals that
mathematical models obtained from the central composite design
3.5. Bead morphology
were well tted.
The morphological analysis of calcium alginate/gum Arabic
3.3. DEE beads containing glibenclamide was visualized by SEM at different
magnications and is presented in Fig. 3(a and b). The SEM photo-
The DEE (%) of all these calcium alginate/gum Arabic beads graph of these beads showed spherical shape with a rough surface.
containing glibenclamide was within the range, 54.17 1.54 to Detailed examination of the bead surface topography revealed
73.93 3.37% (Tables 1 and 4). It was observed that DEE (%) of these cracks and wrinkles, which might be caused by partly collapsing
beads containing glibenclamide was increased with the increment the polymeric gel network during drying [34]. However, polymeric
of sodium alginate and gum Arabic amount in the polymer-blend. debris which was seen on the bead surface could be due to the
The increased DEE (%) with the increasing amount of sodium algi- method of preparation (i.e., simultaneous gel bead preparation and
nate and gum Arabic in these beads may be due to the increase formation of the polymer blend matrix).
in viscosity of the polymer solution with the increasing amount of
polymer addition, so that, it might have prevented drug leaching
to the cross-linking solution. In addition, the increasing amount of Table 5
sodium alginate and gum Arabic in polymer-blend might have ele- Mean diameter of calcium alginate/gum Arabic beads containing glibenclamide.
vated the cross-linking by CaCl2 through availing more numbers of Formulation codes Mean diameter (mm)a
sites for ionic cross-linking.
F-1 1.18 0.11
F-2 1.32 0.13
3.4. Bead size F-3 1.39 0.16
F-4 1.50 0.10
F-5 1.15 0.11
The average size of these formulated dried beads containing
F-6 1.51 0.18
glibenclamide ranged from 1.15 0.11 to 1.55 0.19 mm (Table 5). F-7 1.32 0.16
Increasing the bead size was found with the increasing amount of F-8 1.20 0.12
the polymers, sodium alginate and gum Arabic into bead formula- F-9 1.24 0.13
tions containing glibenclamide. This could be attributed due to the F-O 1.55 0.19

increase in viscosity of the polymer blend (sodium alginate and gum a

Mean S.D.
1076 A.K. Nayak et al. / International Journal of Biological Macromolecules 51 (2012) 10701078
Fig. 3. Scanning electron microphotograph of optimized calcium alginate/gum Arabic beads containing glibenclamide (F-O) (a and b).
3.6. FTIR spectroscopy
The FTIR spectra of sodium alginate, gum Arabic and calcium
alginate/gum Arabic beads without drug, calcium alginate/gum
Arabic beads containing glibenclamide, and pure glibenclamide
are shown in Fig. 4. The FTIR spectra of sodium alginate showed
the band around 30003600, 1614, 1417 and 1032 cm1 , which
are due to the stretching of OH, COO (asymmetric), COO
(symmetric), and C O C, respectively. In the FTIR spectrum of
gum Arabic showed characteristic peaks around 30003600 cm1
for OH stretching, 2993 and 2918 cm1 for CH stretching and
1612 cm1 for C O stretching. The FTIR spectrum of calcium algi-
nate/gum Arabic beads without drug showed characteristic peaks
of both sodium alginate and gum Arabic without any signicant
shifting or deviations. In the FTIR spectrum of pure glibenclamide,
the principal absorption peaks appeared at 3314 cm1 due to the
NH stretching, 3116 cm1 for aromatic hydrogen absorption, and
a peak at 1717 cm1 occurred due to C O absorption peak. In
the FTIR spectrum of calcium alginate/gum Arabic beads containing
glibenclamide, various characteristic peaks of sodium alginate, gum
Arabic, and glibenclamide appeared without any signicant shifting
or deviation of these peaks. In short, the calcium alginate/gum Ara-
bic beads containing glibenclamide prepared by ionotropic gelation
method had signicant characters of glibenclamide in the FTIR
spectrum, suggesting, there was no interaction between the drug,
glibenclamide and the polymers used (sodium alginate and gum
Arabic).
Fig. 4. The FTIR spectra of sodium alginate (a), gum Arabic (b), calcium alginate/gum
Arabic beads without drug (c), calcium alginate/gum Arabic beads containing gliben-
clamide (F-O) (d), and pure glibenclamide (e).
3.7. In vitro drug release
The in vitro glibenclamide release studies were carried out
for various calcium alginate/gum Arabic beads containing gliben-
clamide in the 0.1 N HCl (pH, 1.2) for rst 2 h and then, in phosphate
buffer (pH 7.4) for next 5 h. All these calcium alginate/gum Arabic
beads showed prolonged release of glibenclamide over 7 h (Fig. 5).
Glibenclamide release from these beads in the acidic medium was
less than 15% after 2 h for all these beads containing glibenclamide
due to the shrinkage of pH sensitive alginate at acidic pH. The trace
amount of glibenclamide release in the initial period could prob-
ably be due to the surface adhered drug crystals. After that, drug
release was observed faster in phosphate buffer (pH 7.4) compar-
atively. This might be due to the higher swelling rate of alginate in
phosphate buffer. The percentage drug released from calcium algi-
nate/gum Arabic beads containing glibenclamide in 7 h (R7 h , %) was
within the range of 35.68 1.3853.23 2.22%, and was found to
be lower with the increasing of the polymers, sodium alginate and
gum Arabic, present in the polymer-blend. In case of beads con-
taining higher polymer content, the more hydrophilic property of
the polymers could probably bind better with water to form vis-
cous gel-structure, which might blockade the pores on the surface
of these beads and sustain the drug release for prolonged period.
The in vitro drug release data from various calcium algi-
nate/gum Arabic beads containing glibenclamide were evaluated
kinetically using various mathematical models like zero order,
rst order, HixsonCrowell, Weibull, BakerLonsdale, Higuchi, and
KorsmeyerPeppas models. The R2 values of these models were
determined for evaluation of accuracy and prediction ability of
these models using KinetDS 3.0 Rev. 2010 software. The result
of the curve tting into various mathematical models is given in
Fig. 5. In vitro drug release from various calcium alginate/gum Arabic beads con-
taining glibenclamide [mean S.D., n = 3].
 A.K. Nayak et al. / International Journal of Biological Macromolecules 51 (2012) 10701078 1077

Table 6
Results of curve tting of the in vitro glibenclamide release data from different calcium alginate/gum Arabic beads.

Models Formulation codes

F-1 F-2 F-3 F-4 F-5 F-6 F-7 F-8 F-9 F-O

Zero order R2 0.9347 0.9073 0.9218 0.8985 0.9237 0.9166 0.9011 0.9434 0.9214 0.9051
First order R2 0.9985 0.9904 0.9955 0.9886 0.9953 0.9946 0.9876 0.9957 0.9895 0.9900
HixsonCrowell R2 0.9875 0.9737 0.9804 0.9672 0.9815 0.9797 0.9670 0.9878 0.9771 0.9715
Weibull R2 0.8691 0.8442 0.8508 0.8271 0.8531 0.8547 0.8256 0.8707 0.8431 0.8413
BakerLonsdale R2 0.7576 0.7594 0.7797 0.7570 0.7790 0.7680 0.7642 0.8082 0.7768 0.7662
Higuchi R2 0.6539 0.5750 0.6130 0.5883 0.6196 0.5837 0.6052 0.6622 0.6250 0.5417
KorsmeyerPeppas R2 0.8970 0.8654 0.8759 0.8492 0.8787 0.8772 0.8452 0.8957 0.7679 0.8567
na 0.73 0.77 0.75 0.74 0.75 0.79 0.72 0.74 0.73 0.81
a
Diffusional exponent.

Table 6. When the respective R2 of calcium alginate/gum Arabic
beads containing glibenclamide was compared, it was found to fol-
low the rst-order model (R2 = 0.98860.9985) over a period of
7 h. The value of release exponent (n) determined from in vitro
glibenclamide release data of various calcium alginate/gum Arabic
beads ranged from 0.72 to 0.81, indicating anomalous (non-Fickian)
diffusion mechanism for drug release. The anomalous diffusion
mechanism of drug release demonstrates both diffusion controlled,
and swelling controlled drug release from calcium alginate/gum
Arabic beads containing glibenclamide.
3.8. Swelling behavior
The swelling behavior of optimized calcium alginate/gum Ara-
bic beads containing glibenclamide was evaluated in 0.1 N HCl, pH
1.2, and phosphate buffer, pH 7.4. The swelling behaviors of these
beads in both the pH, 0.1 N HCl (pH 1.2), and phosphate buffer (pH
7.4) are shown in Fig. 6. The swelling index of optimized calcium
alginate/gum Arabic beads containing glibenclamide was initially
lower in acidic pH (0.1 N HCl, pH 1.2) in comparison with that of in
alkaline pH (phosphate buffer, pH 7.4), indicating a pH-sensitive
swelling behavior. This might have occurred due to shrinkage
of alginate at acidic pH. Maximum swelling of these beads was
noticed at 12 h in phosphate buffer, pH 7.4 and after which, ero-
sion and dissolution of swollen cross-linked alginate/gum Arabic
beads took place. The swelling behavior of optimized calcium algi-
nate/gum Arabic beads in alkaline pH could be explained by the
ion exchange phenomenon between calcium ions, which are in
binding with carboxylic groups of cross-linked alginate/gum Arabic
matrix and the sodium ions present in phosphate buffer due to the
inuence of calcium-sequestrant phosphate ions. This argument
may be explained by the observation of some turbidity, which was
appeared in the phosphate buffer might be due to the formation
of calcium phosphate on the same analogy as mentioned by Pas-
parakis and Bouropoulos [34]. Finally, the alginate beads begin to
disintegrate, when calcium ions in the egg-box buckled structure
diffuse out into the swelling medium. Similar ion exchange process
may occur in case of calcium coordinated carboxylic groups present
in the gum Arabic chain. These results clearly suggested that the
optimized calcium alginate/gum Arabic beads containing gliben-
clamide might have the capability to swell slightly in the stomach
at acidic pH as they subsequently move to the upper intestine and
swell more in the intestinal alkaline pH.
4. Conclusion
Calcium alginate/gum Arabic beads containing glibenclamide
were successfully prepared by ionotropic gelation method and
optimized using central composite design. These optimized cal-
cium alginate/gum Arabic beads are excellent combination of high
drug encapsulation efciency (86.02 2.97%), and suitable sus-
tained drug release pattern over prolonged period (cumulative drug
release after 7 h of 35.68 1.38%), which could possibly be advan-
tageous in terms of advanced patient compliance with reduced
dosing interval. The technique for the preparation of calcium algi-
nate/gum Arabic beads containing glibenclamide was found to be
simple, reproducible, easily controllable, economical and consis-
tent. Besides, the excipients such as sodium alginate, gum Arabic
and calcium chloride used for the formulation of these beads were
cheap and easily available.

Acknowledgements

Authors are thankful to Principal and Management of Seemanta

Institute of Pharmaceutical Sciences, Mayurbhanj-757086, India for
providing necessary facilities.

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