ISSN 1061-933X, Colloid Journal, 2006, Vol. 68, No. 1, pp. 115119. Pleiades Publishing, Inc., 2006.

Original Russian Text Yu.L. Khromova, 2006, published in Kolloidnyi Zhurnal, 2006, Vol. 68, No. 1, pp. 123128.

115

INTRODUCTION
In recent years, homogeneous calcium alginate gels
have found wide use in biotechnology [1, 2], pharma-
cology [3, 4], and implantology [5, 6]. This is caused by
their unique combination of physicochemical proper-
ties, biocompatibility, as well as availability and rela-
tively low prices of their components. The main com-
ponent of the gels is alginate, a natural polysaccharide
mainly isolated from brown algae, where its content
can reach 40% of the dry weight of the plant. The algi-
nate molecule consists of

-L-guluronic and

-D-man-
nuronic acid residues connected by

1 4

glycosidic
bonds. The monomer distribution in the alginate chain
corresponds to block copolymers, because there are
polyguluronic (G-blocks) and polymannuronic (M-
blocks) fragments and alternating residues of both
acids in the molecule. The supramolecular structure of
G-blocks of the alginate molecule has a folded confor-
mation, whereas M-blocks form striplike structures.
The formation of calcium alginate gels has been
studied in detail [712]. It is based on strong linking
between Ca

2+

and carboxyl groups of G-blocks of adja-
cent alginate molecules when calcium salt is introduced
into the gelling solution. The cation size and the
supramolecular structure of G-blocks both favor subse-
quent coordination of Ca

2+

with six oxygen atoms of
alginate OH groups located in G-blocks in close vicin-
ity of the cation. This factor causes additional strength-
ening of the gel structure. The process itself has a pro-
nounced cooperative character because the bond for-
mation between Ca

2+

and adjacent alginate molecules
launches the linking of the next chain portion by other
cations. Because of this, the gelling reaction occurs at a
high rate, leading to nonuniform crosslink density and,
as a consequence, to inhomogeneous structure of the
material. Therefore, the principal difculty in the prep-

aration of homogeneous gels is to provide homoge-
neous distribution of cations in the gelling solution.
Presently the most promising method for prepara-
tion of homogeneous gels of calcium alginate is based
on the use of poorly soluble calcium salts, such as car-
bonate, sulfate, phosphate, or hydrophosphate, whose
dissolution provides slow supply of Ca

2+

ions to algi-
nate molecules [2, 4, 9, 10, 1216]. Despite the poor
solubility of Ca

3

(

PO

4

)

2

and CaHPO

4

, the release rate of
calcium cations remains high; hence, in order to create
optimal reaction conditions, passivators are added, such
as pyrophosphate or polyphosphate. In the case of
CaCO

3

, the addition of D-glucono-

-lactone leads to
slow acidication of the medium and thus promotes
gradual dissolution of the salt and release of calcium
cations into the gelling solution. However, the presence
of one or another salt or additive can restrict the eld of
gel application; therefore, it is necessary to improve the
system composition.
In this work, a modied method for preparation of
homogeneous calcium alginate gels using calcium sul-
fate is suggested. This approach is characterized by
introduction of KCl, NaCl, or NH

4

Cl into the gelling
system in order to gradually increase the CaSO

4

solu-
bility. The mechanical and absorption properties of the
gels obtained using KCl are studied, and the possible
mechanism of chloride action on the formation of gel
structures is suggested.
MATERIALS AND METHODS
In order to prepare the gels, average-viscosity
sodium alginate isolated from

Macrocystis pyrifera

(ICN Biomedical, United States) was used. Calcium
sulfate, potassium chloride, sodium chloride, and

The Effect of Chlorides on Alginate Gelation
in the Presence of Calcium Sulfate

Yu. L. Khromova

Institute of Chemistry, Far East Division, Russian Academy of Sciences,
pr. Stoletiya Vladivostoka 159, Vladivostok, 690022 Russia

Received June 21, 2005

Abstract

The preparation of the alginate

CaSO

4

KCl/NH

4

Cl

gel phases is described. Dispersion of calcium
sulfate in solutions with a low chloride content is found to lead to the formation of homogeneous alginate gels.
The dependences of mechanical and absorptive properties of the gels on the chloride concentration are studied,
and a possibility of practical application of the compositions thus prepared is shown. The effect of the KCl and
NH

4

Cl additives on gelation is explained by attainment of the optimal rate of CaSO

4

dissolution, which favors
uniform distribution of Ca

2+

cations in the bulk of the arising gel structure.

DOI:

10.1134/S1061933X06010157

116

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KHROMOVA

ammonium chloride were of pure grade. Distilled water
was prepared by the standard procedure.
Calcium alginate gels were prepared by mixing a
sodium alginate solution and a weighed amount of
CaSO

4

dispersed in a KCl, NH

4

Cl, or NaCl solution for
5 min. The sodium alginate and calcium sulfate concen-
trations in the resultant compositions were maintained
constant: 1 and 0.1 wt %, respectively. The composi-
tions were intensely stirred for 30 s and put aside after
the beginning of thickening. The formation of the gels
was completed within 13 min. To reach equilibrium,
the gels were stored for 3 days.
The mechanical properties of the homogeneous alg-
inate gels were determined by the dynamic rheology
method on a Rotovisco RT 20 rotary viscometer
(Haake, Germany) at a temperature of

25.00

0.05

C
maintained with a KC 20 thermostat (Haake, Ger-
many). The measurement module was of the coneplate
geometry (cone diameter 60 mm, angle

1

). The gap
between the plate and the cone was equal to 0.052 mm
and set automatically.
The determination of the static viscosity (

0

) and
plateau modulus (

G

0

) was performed in the creep mode
at a constant shear stress, and the strain of the sample
over time was measured in the process (Fig. 1). This
method allows us to separate the viscous and elastic
reactions of the tested sample, because the deforma-
tional changes in this case consist of the elastic strain
and the ow strain. The rst of them immediately arises
when the stress is applied, because it is caused by elas-
tic stretching of the network structure; therefore, the
plateau modulus is determined as [17, 18]

(1)

where

is the sample strain and

is the applied shear
stress.
G
0
/ , =

Further much smaller strain is caused by the sample
ow and characterizes the system viscosity. In the
graph of the sample strain vs. time at constant

, the
curve within this region approaches a plateau. There-
fore, the static viscosity is found from expression

(2)

where

is the slope angle of the strain curve.
In order to study the strength characteristics, the
samples were subjected to the action of gradually
increasing shear stress applied at a frequency of 1 Hz.
In this case, the dynamic storage and loss moduli were
measured, and their sum determines the elastic
response of the system by the equation

(3)

where

G

'

is the storage modulus, corresponding to the
elastic response of the sample, and

G

''

is the loss mod-
ulus, reecting the dissipation of the applied energy due
to ow of the sample.
For gel systems representing viscoelastic materials,
the

G

'

value is always higher than

G

''

in the absence of
loading. This means that the system is more elastic than
viscous. When the stress gradually increases, the sys-
tem has no time to relax and, at a certain loading value,
the condition of equality of the storage and loss moduli
is fullled (Fig. 2). At this point, irreversible disintegra-
tion of the structure begins, leading to ow of the sam-
ple; i.e., under further loading of the sample,

G

''

becomes higher than

G

'

. This stress value corresponds
to the critical shear stress (

cr

) and characterizes the
ultimate strength of the material.
The degree of swelling was determined for samples
dried in a thermostat at

37

for 3 days. The lms pre-
pared in such a manner were immersed into distilled
water for one day. After the excess water was removed,
the hydrogels were weighed, and the degree of swelling

S

was calculated by the formula

(4)

where

m

0

and

m

1

are the weights of the dried and swol-
len samples, respectively.
RESULTS AND DISCUSSION
Transparent homogeneous alginate gels were pre-
pared using KCl and NH

4

Cl solutions at low chloride
concentrations. Throughout the whole experiment, no
phase separation (syneresis), which is typical of algi-
nate gels, was observed. In the case of NaCl, just as in
the absence of chlorides, the systems demonstrated
microphase separation. A similar description of the
phase state of the two-component CaSO

4

alginate sys-
tem is given in [5]. The formation of similar structures
was also observed in samples where the KCl content
was higher than 0.12 mol/l and the NH

4

Cl content was
higher than 0.17 mol/l. If dry salts were used and

0
/ , tan =
G G' iG'' , + =
S
m
1
m
0

m
0
------------------, =

0

, %

t

, s
4000 1000 2000 3000
0.01
0.02
0.03
0.04
0
0.05

/

G

0

tan

=

/

0

Fig. 1.

Strain of the sample containing alginate (1 wt %),
CaSO

4

(0.1 wt %), and KCl (0.05 mol/l) versus time at a
constant shear stress of 1.5 Pa.

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THE EFFECT OF CHLORIDES ON ALGINATE GELATION 117

CaSO

4

and chlorides were mixed in the absence of
water and then the alginate solution was added, hetero-
geneous compositions containing CaSO

4

lumps as
inclusions were obtained.
The inuence of the KCl and NH

4

Cl additives on the
mechanical properties of homogeneous calcium algi-
nate gels can be evaluated from the dependences of the
static viscosity, plateau modulus, and critical shear
stress on the chloride content. Variations in the NH

4

Cl
concentration do not signicantly affect the rheological
parameters of gels (graphs are not shown). However,
note that the mechanical properties of the gels prepared
using NH

4

Cl were lower than those of the gels prepared
using KCl.
The dependence of the static viscosity of calcium
alginate gels on the KCl content is shown in Fig. 3a. It
is seen that the addition of the salt leads to linear
increase in

0

. This indicates increasing interaction
between macromolecules of the gel network structure
with increasing KCl concentration in the system. The
increase in the plateau modulus values (Fig. 3b) reects
increasing structural density due to the growing number
of crosslinks between alginate molecules through Ca

2+

ions [19, 20]. For chloride concentrations higher than
0.05 mol/l, the interaction intensity signicantly
decreases, indicating uniform inclusion of a larger
amount of calcium cations in the gel structure.
Note that the values obtained for the rheological
parameters are in a complete agreement with the data of
earlier works where the properties of homogeneous alg-
inate gels based on CaCO

3

and the D-glucono-

-lactone
additive were studied [2123]. Under similar condi-
tions (CaCO

3

concentration 0.1 wt % and alginate con-
centration 1 wt %), the static viscosity and the plateau
modulus values are about

10

5

Pa s and

10

2

Pa, respec-
tively.
The dependence presented in Fig. 4 reects the vari-
ations in the critical shear stress with increasing KCl
content. The

cr

value is an important characteristic that
allows one to evaluate the material strength. As is seen
from the graph, the curve passes through a maximum,
i.e., the gel strength initially increases, and then, with
increasing KCl concentration (>0.05 mol/l), the gels
become more fragile. If it is taken into account that

cr

directly depends on the supramolecular structure, the
increasing number of crosslinks between macromole-
cules (Fig. 3b) must favor strengthening of the gels.
This contradiction can be resolved if one supposes that
the presence of K

+

and Cl

ions can affect the properties
of the network structure and, at high chloride concen-
trations, lead to increasing gel fragility under deforma-
tion.
The swelling capacity allows wide use of alginate
gels as absorbents in hygienic products [4] and in treat-
ment of wounds [3, 24]. Figure 5 represents the degree
of swelling of the dried gel samples as a function of the
KCl concentration in the system. It is seen that an
increasing chloride content leads to a sharp drop in the
absorbing capacity of calcium alginate gels. However,
note that the swelling degree of the gels is 40 even at the
highest KCl concentration; i.e., 1 g of the dried gel is
capable of absorbing up to 40 g of water. According to
[4, 25], the swelling capacity is determined by the sum
of three factors: (1) nonuniform distribution of counte-
rions inside and outside the gel, (2) osmotic pressure
arising during mixing in the polymersolvent system,
and (3) capacity of the network structure for elastic
stretching. In this connection, the decreased absorption
of water by alginate gels containing KCl can be

20 10 30 40
300
200
100
400

G

''

G

'

cr

, Pa

G

',

G

'', Pa
Fig. 2. Determination of critical shear stress
cr
for the sam-
ple containing alginate (1 wt %), CaSO
4
(0.1 wt %), and
KCl (0.05 mol/l) (see text for explanation).
0.08 0.12 0.04 0
3
4
2
1
(a)
KCl concentration, mol/l

0
10
6
, Pa s
0.08 0.04 0.12 0
100
120
140
80
60
40
(b)
G
0
, Pa
Fig. 3. (a) Static viscosity
0
and (b) plateau modulus G
0
of
calcium alginate gels versus KCl content. The alginate and
CaSO
4
concentrations are 1 and 0.1 wt %, respectively.
118
COLLOID JOURNAL Vol. 68 No. 1 2006
KHROMOVA
explained by the chloride action on the elastic proper-
ties of the supramolecular network. As follows from the
behavior of the plateau modulus (Fig. 3b), the addition
of KCl leads to increasing crosslink number in the net-
work structure. This restricts the mobility of fragments
of alginate macromolecules and provides the determin-
ing factor for the decrease in the degree of gel swelling.
In order to explain the action of the KCl and NH
4
Cl
additives on the formation of homogeneous calcium
alginate gels, we will consider the possible mechanism
of gelation in the presence of these chlorides. As was
previously found, the formation of the gel network
structure occurs owing to binding of bivalent calcium
ions with G-blocks of adjacent alginate molecules [11,
12, 14, 16]. Thus, univalent K
+
, , and Cl

ions do
not participate in the crosslinking. The action of KCl
and NH
4
Cl on alginate gelation is primarily explained
by their inuence on the behavior of calcium sulfate in
solution. The presence of chlorides is known to
increase the CaSO
4
solubility owing to the salting-in
effect. In this case, a shell of chloride ions, which
reduces the reactivity and mobility of calcium ions, is
formed around each Ca
2+
ion. During alginate gel for-
mation, intense stirring provides uniform distribution
of cations within the bulk before the G-blocks of algi-
nate molecules begin to interact with Ca
2+
cations. The
difference in the inuence of KCl and NH
4
Cl on the gel
properties is likely to be explained by their action on the
formation of the supramolecular network structure. The
K
+
and ions are known to be largely similar to
each other, with the principal difference in the fact that
NH
4
+
NH
4
+
ammonium ions can form hydrogen bonds. On this
basis, one can suppose that ions, being involved
in the network of hydrogen bonds stabilizing the gel
structure, hinder close contacts between Ca
2+
and
OH-groups of alginate G-blocks. This is indicated by
lower rheological parameters of such gels compared to
compositions with the KCl additive.
Microphase separation at KCl concentrations higher
than 0.12 and NH
4
Cl concentrations higher than 0.17
mol/l can be explained by two factors. The solubility of
CaSO
4
linearly increases with the chloride concentra-
tion, and the formation of crosslinks in the alginate gel
occurs at a high rate. In this connection, despite the
intense stirring, a sharp increase in the concentration of
Ca
2+
ions leads to their nonuniform distribution within
the solution. As is suggested in [26] (this study con-
cerns copper alginate gels containing NaCl as an addi-
tive), an additional factor in the formation of
microphases can be a decrease in the charge density on
polysaccharide molecules resulting from electrostatic
interactions between the K
+
or ions and alginate
COO

groups. Owing to this, the coilglobule confor-

mational transition occurs in the solution, leading to
prevalence of intramolecular interactions over intermo-
lecular ones.
In conclusion, one can state that the inuence of the
chlorides studied on the formation of homogeneous
alginate gels consists in their action on the dissolution
of CaSO
4
. Within the studied concentration range,
potassium chloride and ammonium chloride provide
the optimal content and supply rate of calcium cations
NH
4
+
NH
4
+
0.08 0.12 0.04 0
40
50
30
KCl concentration, mol/l

cr
, Pa
Fig. 4. Critical shear stress of calcium alginate gels versus
KCl content. The alginate and CaSO
4
concentrations are 1
and 0.1 wt %, respectively.
0.04 0.08 0.12
40
60
80
100
120
S
KCl concentration, mol/l
0
Fig. 5. The degree of swelling of calcium alginate lms ver-
sus KCl content. The alginate and CaSO
4
concentrations
are 1 and 0.1 wt %, respectively.
COLLOID JOURNAL Vol. 68 No. 1 2006
THE EFFECT OF CHLORIDES ON ALGINATE GELATION 119
to alginate macromolecules, whereas intense stirring
provides their more uniform distribution within the
forming gel structure.
The results of this work indicate that the use of cal-
cium alginate gels containing KCl and NH
4
Cl as addi-
tives is promising for practical applications. Particular
attention shows be paid to systems obtained with the
use of KCl because of their high mechanical and
absorption properties. Another important advantage of
these compositions is simplicity of their preparation
and biocompatibility of all the gel components.
ACKNOWLEDGMENTS
This work was supported by the Far East Division of
the Russian Academy of Sciences (youth grant).
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