Food Hydrocolloids Vol. 11 no. 2 pp.

159-170, 1997
-
Multicomponent biopolymer gels

D.Y.Zasypkin 1,3, E.E.Braudo 1 and Y.B.'Iolstoguzev-

llnstitute of Biochemical Physics, Russian Academy of Sciences, Vavilov Str. 28, 117813 Moscow GSP-1, Russia and 2Nestle
Research Centre, Vers-chez-Ies-Blanc, PO Box 44, CH 1000 Lausanne, Switzerland
3To whom correspondence should be addressed

Abstract
The structure-property relationship and application of multicomponent protein-polysaccharide and
polysaccharide l-spolysaccharide 2 gels are considered. Attention is focused on gels based on mixed
solutions of gelatin with dextran, human serum albumin, sodium caseinate, ovalbumin, agarose,
methylcellulose, calcium alginate or sodium alginate. The thermodynamic incompatibility of and the
formation of complexes by food hydrocolloids greatly affect mechanical and other physicochemical
properties of gels. Food hydrocolloid mixtures can display both synergistic and antagonistic effects.
Factors such as chemical structure, conformational state and molecular weight of hydrocolloids, as well
as the composition of their mixed solution, p H, ionic strength and temperature, are of key importance
for the structure of multicomponent gels.

Introduction
Increasing attention has been focused on the gelation of
mixed aqueous solutions of proteins and polysaccharides
during the last three decades (I-IS). This trend reflects (i) a
key role of hydrocolloids in structure formation in foods,
(ii) an increasing number of hydrocolloids used as food
additives, e.g. xanthan, gellan, concentrated or isolated
proteins from soybean, milk whey proteins, beta-lacto-
globulin, gluten, as well as (iii) the development of new food
processing methods, such as microwave heating, thermo-
plastic extrusion (11,16) and high-pressure treatment (17,18).
It should be noted that several important findings in the field
of non-specific hydrocolloid interactions in aqueous media,
such as interbiopolymer complexing and thermodynamic
incompatibility of hydrocolloid solutes of ternary systems
(1-12), have been extensively elucidated.
Hydrocolloids are mainly responsible for the functional
properties of processed food systems and the quality of
many foods. Formulated foods usually contain mixtures of
hydrocolloids performing structural functions. Investiga-
tions of hydrocolloid interactions in aqueous solutions and
gels are necessary for modelling and improvement of
conventional foods, the development of novel formulated
foods and for controlling functional properties of food
systems by added hydrocolloids (1-3) . However, in spite of
the wide practical use of hydrocolloids as food additives
and ingredients, many fundamental aspects of the
structure-property relationship of mixtures of gel-forming
hydrocolloids in liquid solutions and gels were until recently
unknown .
The main aims here are to consider: (i) general features of
multicomponent gels (1-14,19-23) and (ii) some findings
made by the Laboratory of Novel Food Forms, USSR
Academy of Sciences, Moscow. This was the first group to
start investigations in the field of mu1ticomponent
biopolymer solutions and gels (1-7). The fundamental
research programme on formulated foods was initiated by
Professor A.N.Nesmeyanov (IS). We will only consider
gelation of mixed solutions of two different biopolymers,
which do not contain colloidal particles.
Classification of multicomponent gels containing
two biopolymers
Gels formed by several gelling agents can be divided into
three groups, namely filled, mixed and complex gels
(1-3,6,9-13,27). This classification is rather similar to that
proposed by VMorris (8).
1. Filled gels containing one or several independent
biopolymer networks forming a continuous phase filled
by dispersed particles. The latter can be of a different
chemical nature and state of matter, i.e. solid, liquid and

e OXford University Press

160 D. VZasypkin, E.E.Braudo and VB. Tolstoguzov
gas. A biopolymer network can contain one or several
biopolymers, Accordingly, filled mixed and filled complex
gels can be distinguished. There are two other types of
filled biopolymer gels (6). Dispersed particles of a
biopolymer gel may be macromolecules of biopolymers
and/or colloidal particles of biopolymers. This means that
a filled biopolymer gel may be either a single-phase or a
two-phase gel. A single-phase biopolymer gel contains a
biopolymer network and other biopolymer (filler) in the
form of molecules or molecular aggregates of the solute.
Two (or more)-phase biopolymer gels consist of a
continuous network filled with dispersed particles.
According to V Morris (8), filled gels are regarded as
phase-separated gels.
2. Mixed gels consisting of two (or more) independent
networks. Both networks span the entire system, but any
interaction between them is mostly topological.
According to Y.Morris (8), mixed gels have been named as
gels with interpenetrating networks. Because thermo-
dynamic incompatibility of biopolymers is of a general
nature and typical of foods (1-3,7,25,26), it can be
assumed that the formation of interpenetrating networks
is quite seldom a particular case of mixed gels. Such
unique gel-like systems will be considered below using
gelatin-calcium alginate gels as an example. Gelation of
the phase-separated system, i.e. water-in-water emulsion
containing two limitedly co-soluble gel-forming biopoly-
mers, can lead to a filled gel, both phases of which are also
filled gels.
3. Complex gels [coupled networks, according to Y.Morris
(8)] involving direct association between the two polymers
and subsequent network formation.
A restricted availability of information about
multicomponent gel structures makes it difficult to apply
both kinds of gel classification.
Gels filled with active molecular and/or colloidal fillers, i.e.
fillers having an affinity to the gel-forming biopolymer
making up the network, can be considered as complex gels. A
mixed gel containing two continuous phases-the disperse
phase (formed by several biopolymer three-dimensional
networks) and the dispersion medium (which is a biopolymer
solution)-can also be regarded as a particular case of filled
gels. The gels comprising two biopolymer gelling agents can
be referred to as mixed gels when no clear border exists
between the different types of gel. This most general
terminology will be used below. Other terms will be used
where a more strict definition of multicomponent gel
structure is necessary.
Structure-property relationship of multicomponent
gels. Influencing physicochemical factors
Several physicochemical phenomena govern and make
predictable the formation of gel structures in
multicomponent solutions.
Excluded volume effects in single-phase mixed biopolymer
solutions and gels
Molecules of different biopolymers cannot occupy the same
volume in the bulk of their mixed solution when there is no
specific interaction between them. This means that
incompatible biopolymers mutually concentrate each other
in solution. As a result of the excluded volume effect, the rate
of gelation and elasticity modulus of mixed gels can be
higher and the minimum concentration for gelation is lower
(11-13,26-28).
Phase separation of a mixed solution of two biopolymers
Normally, mixed solutions of chemically and/or
conformationally different biopolymers are unstable and
separate into two liquid phases. This occurs at a certain
sufficiently high bulk concentration of the biopolymers.
Biopolymers are concentrated in different phases and form a
water-in-water emulsion.
The chemical analysis of the phases is usually presented in
the form of a phase diagram.
Boundary conditions for the formation of various types of
gels presented in Figure 1 are mainly determined by two
characteristics of mixed solutions of biopolymers
(13,25-27). These are the co-solubility of the biopolymers
and the minimum concentrations for gelation of each of
them in the presence of the other one. This approach is
limited by the absence of information about the effects of
heating and gelation on the phase equilibrium in the
phase-separated biopolymer system. For this reason, in
practice, the structure of multicomponent gels is usually
studied by physicochemical analysis, i.e. by the system
property-composition used relationship.
Hydrocolloids can form mixed gels when the
concentration of each of them in the mixed solution exceeds
the minimum concentration for gelation. Normally, the
minimum concentration for heat-induced gelation of
biopolymers varies from 0.1 to 15% w/w. For instance, the
gel-point is -0.1% w/w for agarose, -1 % w/w for anionic
polysaccharides and gelatin, exceeds 6% w/w for beta-
lactoglobulin and lIS soybean globulin, and 13% w/w for the
II S broad bean globulin. The minimum concentration for
gelation usually decreases when another incompatible
biopolymer is added. This is due to an excluded volume effect
in the single-phase mixed solution. In the phase-separated
biopolymer systems, the same effect may be due to water
redistribution between phases during gelation.
The range of minimum concentrations for hydrocolloid
gelation is usually lower than that of the phase separation
threshold (27). For instance, the phase separation threshold
is -2-4% w/w for mixtures of gelatin with linear anionic
polysaccharides and exceeds 12% w/w for solution mixtures
of globular proteins (11-13,27).
This means that a mixture of biopolymer gelling agents
can form gels in concentration areas lying both above and
below the binodal curve. Accordingly, gelation of the mixed
 Multicomponent biopolymer gels 161

-c:::n
c (0)
above) sides of the binodal curve when the binodal curve is
located above the curve (shown by points) of minimum
Q) concentration for gelation. The points (below the binodal
c:::n
I curve) correspond to the transition from single-phase filled
C
o gels (one network) to single-phase mixed gels (two networks) .
.s,
In other words, the position of the curve of minimum
concentration for gelation of a biopolymer mixture deter-
mines the fine structure (one or two networks) of gels.
"" C" 2
". 2 Distribution of water between co-existing phases of a gel
Polymer P2 (gelling) E.R.Morris (29) suggested a method of evaluating water
distribution between phases from the data on mechanical
3 properties of two-phase gels.
---. ( b)
en Water can be redistributed between the phases in the
c
course of going over of two co-existing phases into a
-c:::n
Q)
position of equilibrium (11-13,30,31). Concentration of one
---
o,
of the co-existing phases may be accompanied by its gelation
..... (30,31). Phase separation accompanied by water re-
Q)
E distribution can favour either a decrease or an increase in the
>- elasticity modulus of mixed gels (11-13,26-28). This
0
o,
corresponds either to non-gelling of a dispersed phase and
~ C" 2
~ 2 low adhesion between filler and gel matrix or the
Polymer P2 (gelling) concentration of the continuous phase of the gel (27). When
the continuous phase containing the stronger gelling agent is
Figure 1 Phase diagrams of solvent/polymer I1polymer 2 systems concentrated, a synergistic increase in elasticity modul us of
consistingof: (a) non-gellingpolymer (PI) and gelling polymer (P2); the gel can occur.
(b) two gelling polymers. Cl* and C2* are the minimum
concentrations for the gelation of polymer I and polymer 2, Phase inversion in water-in-water emulsions
respectively. Solid line, binodal curve; thin lines, tie lines; pointed
line, minimum concentration for gelation; dotted line, rectilinear Water redistribution between the phases is accompanied by
diameter. Areas: I, single-phase solution; 2, single-phase gel of P2, an alteration in phase volume ratio. Phase inversion can
filled with PI; 3, single-phase gel of PI, filled with P2; 4, 5, occur in the vicinity of the rectilinear diameter where
emulsions; 6, gel rich in P2, filled with liquid particles rich in PI; 7, co-existing phases have similar volumes. Phase inversion can
suspension: dispersion medium rich in Pl, gel particles rich in P2; 8, lead to a great change in the composition of the continuous
9, gelsfilled with gel particles, each phase being a filled gel. phase and in mechanical properties of the gel (27) . For
instance, an increase in the concentration of the continuous
solution leads to filled gels above the binodal and to phase rich in a relatively strong gelling agent can lead to
single-phase mixed gels below the binodal (Fig. la and b) . phase inversion and to weakening of the gel with the
Figure I illustrates the formation of different gels by continuous phase rich in a relatively weaker gelling agent.
hydrocolloid mixtures when the binodal curve and the curves
of minimum concentration for gelation of the biopolymers Distribution of ions and biopolymer fractions between
are not intersected (28). Two kinds of hydrocolloid mixtures co-existing phases
(6,9,28) containing one (Fig. la) and two gelling agents (Fig.
Ib) can be distinguished. Presumably, in the first case the Partitioning of cations and system components of low
binodal separates single-phase mixed gels from two-phase molecular weight between co-existing phases reflects their
filled gels. The rectilinear diameter probably corresponds to a affinity for the biopolymers forming the phases. Phase
phase inversion in the two-phase gels. As a result, two separation in mixed biopolymer solutions can also be
different gels are obtainable within areas 4 and 5. The accompanied by the fractionation of biopolymers, e.g.
rectilinear diameter separates gels (area 6) containing a polysaccharides (27) . Gelation conditions and properties of
continuous phase rich in biopolymer 2 filled with a gels of many proteins and polysaccharides greatly depend on
liquid-dispersed phase rich in biopolymer I from the presence of certain cations, then fraction composition
suspensions (area 7) containing gel particles rich in and molecular weight. For instance, many anionic
biopolymer 2. polysaccharides are capable of forming gels in the presence
Figure Ib illustrates the formation of gels of different of certain metal ions. Accordingly, the distribution of many
structures based on systems containing two gelling agents. system components can greatly affect the properties of
Formation of gels would be possible on both (below and multiphase gels (27).
162 D. V'Z asypkin, E.E.Braudo and VB. Toistoguzov

Some other effects occurring in mixtures of biopolymers 0.6

(0 )
Other effects of polysaccharides can occur on gelation of
single-phase mixed protein/polysaccharide solutions.
ill
Gel formation can be considered as a partial unfolding of
globular proteins, followed by primary aggregation of these $2
x
unfolded protein molecules and network formation as a QJ
o
C
result of interaction of the primary aggregates (12,13). o
Polysaccharides can affect aggregation of protein molecules n.
E
(13,25). Since gel network formation is a diffusion-controlled o
u
process, the viscosity of the medium is an influential factor.
The size of the primary protein aggregates increases and 2.5
0.5 1.5
the rate of their diffusion and secondary aggregation

.---
Dex-rnn , %, w/w
decreases with an increase in polysaccharide concentration in
0 ...<>-0--0 0 0- 1
single-phase globular protein/polysaccharide solutions. This ._ 2
results in an increased heterogeneity of the gel network
formed and in a weakening of the gel. Further increases in
polysaccharide concentration and in the size of network
structural elements (quite large dispersed particles)
c
o
..-o
-e-
oL..
.
/.
--.-----
......
(~

contribute to an increase in plasticity and spread-like texture o

o
of the gel. +o,
Another effect of biopolymer gelation arises from the fact o
that gelation is always a result of aggregat ion of
macromolecules. Accordingly, gelation is accompanied by a 2 3
strong decrease in the excluded volume effect of I Time, days
macromolecules. This makes the dispersion medium of the 10
0

gel a better solvent for other biopolymers than the initial 3:

.........
\
0
gelating solution (12,13). This also means that the properties 3:,
of a mixed gel depend on the sequence of gelation of (e)
e;e
individual gelling agents in their mixtures (27). c - \
0
5
\
0
L..
+)(
QJ
0 0
Filled gels '0
'------0-0_0 0-
The effect of an added polysaccharide (dextran) on the
formation of gelatin gels is shown in Figure 2 (32,33). The 5 10 15
compliance of gelatin gel depends on the amount of added Gelatin, %, wjw
dextran. Minimum gel compliance corresponds to the
transition from single-phase gelatin gels containing dextran Figure 2 Characteristics of the water/gelatin/dextran system. (a) 30
solution as the dispersion medium to a gelatin gel filled with min compliance of 10% gelatin/de xtran gels versus concentration of
dispersed droplets of dextran solution (Fig. 2a). The minimal dextran of various molecular masses: 1,500 kDa; 2, 65 kDa; 3, 20
value of gel compliance is shifted along the axis of dextran kDa (32). (b) Optical rotation of 4% w/w gelatin gel (curve I) or 4%
w/w gelatin/O.2% w/w dextran (65 kDa ) gels (curve 2) at 6°C versus
concentration with an increase in molecular weight of the
ageing time of gels (32). (e) Phase diagram of water/gelatin/dextran
dextran. (65 kD a) system at 42.5°C (33).
Small quantities of dextran «0.5% w/w provide a
single-phase gelatin gel) greatly increase the rate of gelatin
gelation (Fig . 2b) and the compliance of the gels. The and dextran particles. An increase in concentration of free
compliance of gelatin gel filled with dispersed droplets of the ends of fibrils corresponds to a decrease in the density of the
dextran phase is higher than that of a gelatin gel of the same network (active fibrils) and to an increase in compliance of
concentration. It increases with an increase in the volume the gel.
fraction of the dextran phase and decreases with a decrease Compliance measurement and electron microscopic study
in the average size of droplets of the dextran phase at its of the gelatin gels filled with dextran shows that the phase
constant volume fraction (12,13,32). Electron microscopy separation threshold for the gelatin-dextran-water system
shows that dextran droplets break down the gelatin gel decreases with temperature. Its value for the 10% w/w gelatin
network. Free ends (tails) of gelatin fibrils are normally gel containing 0.3-0.4% w/w dextran at 6°C is lower than
oriented to the surface of the dextran droplets (32). that at 42.5°C obtained from the phase diagram for the
Evidently, this reflects the trend of minimizing the contacts system (Fig. 2a, curve 2 and Fig. 2c, respectively).
between the two phases : the phase containing gelatin fibrils The effect of dextran on the properties of gelatin gels (32)
 Multicomponent biopolymer gels 163

I 150
X

Ol
C
:::
2.0

x
) 35.0
u
0

QI
~
.....
"orn
'-

.....

~~.7I
OJ
~
til
a 1.5 III
x E
0 -t-
Q>
QI
'-
Ol
QJ
a • 30.0
£<lJ
z
L----"'--------'------:-'2S.G
o 5.0 10.0 15.0
Human serum al burnin, % ,w/w
Figure 3 The effect of human serum albumin (HSA) on the melting
temperature of 5% w/w gelatin gel and its equilibrium weight degree
of swelling in 0.5 molldm 3 sodium chloride solution (25). x, degree
of swelling, native HSA, pH 4.8-5.1; 0 , degree of swelling,
denatured HSA, pH 4.8-5.1; e, melting temperature, native HSA,
pH 4.8-5.1 ; _, melting temperature, denatured HSA, pH 4.8-5 .1; D.,
melting temperature, native HSA, pH 8.2. Standard error did not
exceed 0.05 for degree of swelling and O.1°C for the melting
temperature.
is more pronounced than that of globular proteins and
sodium caseinate (24). Figure 3 shows the equilibrium degree
of swelling and melting temperature of 5% (w/w) gelatin
gel containing native human serum albumin. These
measurement were carried out at pH 3.5-5.1, where gelatin
and globular proteins do not form complexes in their mixed
solutions. The addition of up to 7% w/w human serum
albumin did not affect the degree of swelling and melting
temperature of the gelatin gels.
Addition of globular proteins does not change the
relaxation rate of the gelatin gels. Curves of the reversible
creep of 5% w/w gelatin containing 4.5% w/w human serum
albumin can be superimposed on that of the gelatin gel curve
by a shift along the compliance axis in double logarithmic
scale. A decrease in gelatin gel compliance on adding
globular protein may be due to an increase in density of the
gel network rather than to a decrease in the rate of relaxation
of the gels. This increase in density of gel network (of
junction zones) does not affect the equilibrium degree of
swelling of the gel containing a relatively low concentration
of human serum albumin.
Presumably, additional junction zones formed in the
gelatin gel network in the presence of human serum albumin
cannot withstand the osmotic pressure of aqueous medium.
Denaturation of human serum albumin does not lead to the
formation of new effective junction zones in the gel network.
0 u
'-
QI
a.
.....o
c
QI
o
u
3
01 f----- ""'--i
C
/ !
10 30 50
Protein added to gelatin, %, w/w
(toto I protein dry weigh~ c 100%)
Figure 4 Dependence of water contact angle on the content of: I,
human serum albumin (HSA); 2, ovalbumin; 3, casein, in a gelatin
gel (the total protein concentration is 13% w/w) (9).
Figure 3 shows that denaturation of human serum albumin
increases the melting temperature of the gelatin gel but does
not influence the degree of swelling (24). Addition of up to
5% w/w of native or denatured ovalbumin has no significant
effect on the properties of 5% w/w gelatin gels (24).
Figure 3 shows that when the concentration of an added
protein (native human serum albumin) exceeds 7%, the
equilibrium degree of swelling of the gels increases and the
temperature of gel melting decreases. This is probably due to
an increase in size and density of gelatin macromolecular
aggregates that form the gel network. This results in a
decrease in the density of the gel network, i.e. in the effective
number of junction zones of the gelatin gel. The relaxation
rate also rises with the amount of proteins added to the
gelatin gel (24).
Thus, at <7% (w/w) concentration of the globular proteins
in the filled gelatin gels, the above fillers have no significant
effect on degree of swelling, melting temperature or
compliance of filled gels. Respectively, the gelling properties
of the gelatin are not deteriorated by fillers of various
natures. At higher filler concentrations, a marked increase in
the degree of swelling and compliance along with lowering
of the melting temperature are observed. These effects
resulted from a decrease in the number of junction zones in
the filled gels.
Gelatin and human serum albumin can form complexes at
pH 8.2 (34). The formation of gelatin-human serum albumin
complexes leads to a strong increase in the compliance of
gelatin gel, but does not markedly change the temperature of
melting of the gel. Presumably, the competition between
gelatin-gelatin and gelatin-human serum albumin
interactions decreases the density of the gelatin gel network.
Unlike volume properties, the wettability of gelatin gels
strongly depends on the type of added protein. For instance,
164 D. V.Z asypkin, E.E.Braudoand V.B.Toistoguzov

0.8
0-"';;:0- _ _ _

t.-------=-:l---- ,
~ 0.6 ~o I".........~ I °
u
<,
~

~
Y t. I Q)

::J
~

..a......
.
..
c
a
OJ,
Ii
I
\1:1 <ll
a.
E
•
~ 0\
2~:
<ll
<.!l ~ 40
0.2 .~~
_ _ _ _ _ _ _e _
I
t. 0.6 0.2

2 4 6 8
0.05 0.1 0.15 0.2 Agarose, 'Yo, w/w
Agarose . % ,w/w Gelatin , %,Wjw

Figure 5 Minimum concentration for gelation at 20°C versus Figure 7 I, Composition dependence of the melting po ints of
composition of the solventlgelatin/agarose system: 0 , water, pH 4.7; gelatin/agarose hydrogels; 2, composition dependence of the
e, 0.1 moUdm3 sodium chloride, pH 4.7; s ;0.05 moUdm3 disodium low-temperature network melting point of gelatin/agaros e hydrogels
phosphate, pH 6.86 (28). (from thermomechanical dat a). Dotted lines, gel melting po ints of
the corresponding binary polymer/ solvent gels. Gel storage time at
ro-c was I day (28).
& 2.0
",, -
(0) (b) 1.8 r - - - - - - - - - -- - - - - - - ,
I~

" 1.6
III
::l c/°
::l
'0 / 0\
0 ,
E 1.2
01
c .-/
/j I'
I I
C

C1I
~ 1.4

e
::l
. o / // /

0.4 <ll

2 4 6 8
...
C1I
0\
Q)
o

Figure 6 Dependence of the Young modulus of gelatin/agarose

--
0.6 0.2
hydrogels on gel composition at pH 4.7, 20°C. Ageing (at IDoC) after
gelation: 0 , I day; e, 7 days. Dotted lines, Young modulus of the 2 4 6 8
Agarose , % ,w/w
cor responding agarose or gelatin gels. Solvent: (a) water; (b) 0.1
moUdm3 sodium chloride (28). Gelat in, %. w/w

Figure 8 Equilibrium weight degree of swelling of gelatin/agarose

the contact angle of water increases slightly on adding
ovalbumin, probably due to its denaturation at the interface.
The contact angle of a water droplet on the surface of 13%
gelatin gel (on gel/air interface) decreases with an increase in
con centration of human serum albumin and, to a greater
extent, sodium caseinate (9,35).
Mixedgelatin/agarose gels
Figures 5-8 illustrate physicochemical analysis of the phase
state of gelatin/agarose mixed gels (28). Gels were prepared
by swelling of dry blends of biopolymers, followed by
heating in hermetically sealed tubes at 98°C for 30 min and
by cooling and keeping at lOoe for 1-7 days. The
gels at 20°C and pH 4.7 in water (curve I) and in 0.1 moUdm3
sod ium chloride (curve 2). Gel storage time at lQoC was 7 days.
Datted lines, degree of swelling of the corre sponding binary
polymer/solvent gels (28).
temperature of gel melting was determined using a dens er
inert liqu id (37,38). The elasticity modulus was determined
under one-axial compression of gels at constant stress. The
strain corresponded to linear viscoelastic behaviour of the
gels (28).
Figure 5 shows th at the minimum concentration for
gelation of mixed gelatinlagarose solutions was lower than
those of solutions of both individual gelling agents. This is
 Multicomponent biopolymer gels
due to the excluded volume effect of biopolymers in their
mixed solution. Addition of sodium chloride or disodium
phosphate leads to an increase in the minimum concentra-
tion for agarose gelation.
Figure 6 shows the effects of the concentration of the
individual gel-forming agents and the composition of their
mixtures on the elasticity modulus of gels. This figure
illustrates the formation of different structures of multi-
component gels.
Changing the composition of the mixed solution leads to
the transition from single-phase to two-phase gels. A
microstructure study (28) showed that this occurs when 10%
w/w gelatin gel contains from 0.012 to 0.025% w/w agarose.
Phase separation in mixed gelatin-agarose gels containing
1%w/w gelatin and 0.9% w/w agarose, i.e. in the composition
area corresponding to lower level of gelatin, was observed
earlier (36). The spherical shape of the dispersed particles
testifies to a liquid-liquid phase separation taking place in
the mixed solution before its gelation.
The composition-property relationship presented in
Figure 6 is typical of mixed gels (11,27). Small additions of
agarose result in an increase in the elasticity modulus of
single-phase gels due to the excluded volume effect. The same
effect (Fig. 2a) was observed earlier for the gelatin/dextran
gels (32). The position of the maximum of the elasticity
modulus of mixed gels at low agarose concentration does not
depend on gel ageing from 1 to 7 days (Fig. 6a and b). This
reflects an equilibrium nature of this peak. The second peak
is of a non-equilibrium nature. This peak disappears after
more than 1 day storage.
Phase separation creates many disruptions in the gel
network and results in a decrease in the gel modulus
(27,28,32). Because of the relatively higher hydrophilicity of
agarose, phase separation is accompanied by water
redistribution between the two co-existing phases. This leads
to dilution of the dispersed phase rich in agarose and
concentration of gelatin continuous phase, and results in an
additional maximum of the elasticity modulus. The
agarose-rich dispersed particles are an inert filler of gelatin
gels. A further increase in the volume fraction of the
dispersed particles leads to a decrease in the elasticity
modulus of the gel.
The most pronounced minimum of the elasticity modulus
(especially pronounced after 1 day storage of the gel)
corresponds to phase inversion. This latter is confirmed by
the strong change in the temperature of gel melting within
this area of composition of the mixed gel (Fig. 7, curve 1).
There is a difference between the compositions cor-
responding to phase inversion of the mixed gel determined
by different methods, namely from the melting point and the
minimum elasticity modulus of the gel (36,38) and by photon
microscopy (36). This reflects both the effect of loading,
which differs in the three methods used, and the effect of the
composition of the gel continuous phase before phase
inversion on the thermomechanical properties of the gels.
Figure 7, curve 2, shows that in mixed gels with a
165
continuous phase rich in agarose and dispersed particles rich
in gelatin the latter contribute to the elasticity modulus of
the gel and upon heating (at a heating rate of 0.3°C/min)
melt at temperatures close to the melting temperature of the
gelatin gel.
Figure 8 shows the effect of the mixed gel composition on
its equilibrium degree of swelling in both water and 0.1
mol/dm' sodium chloride. The mixed gel always has a higher
degree of equilibrium swelling than that of gels of its
individual biopolymer components at the same con-
centration as in the mixed gel continuous phase. This
probably results from osmotic activity of the second polymer
component. Addition of a biopolymer that does not gel
contributes to an increase in the degree of swelling of gelatin
gels (24,39).
Gelatinlmethylcellulose gels
Gelatin-methylcellulose mixtures were also studied as an
example of a system containing two incompatible
gel-forming biopolymers (9,40,41). Methylcellulose gels on
heating, while gelatin gels on cooling. Figure 9 shows
isothermal phase diagrams of the system at pH 3 and 4.75
(isoelectric point of gelatin) in water and in 0.5 mol/dm!
sodium chloride (40). Phase diagrams were obtained at 35°C,
where neither gelatin nor methylcellulose gel.
Lowering of the pH below the isoelectric point of gelatin
leads to an increase in both the phase separation threshold
and the co-solubility of the biopolymers, i.e. in their
compatibility. Compatibility of these biopolymers increases
only slightly by the addition of 0.5 mol/dm! sodium chloride
at the gelatin isoelectric point. Phase diagrams for mixed
solutions allow the composition-property relationship of
mixed gels to be evaluated. For instance, this relates to water
partition and concentration of the gelatin-rich phase of an
initial mixed solution, as well as the effects of pH and ionic
strength on the phase state of gels.
Gelation of mixed gelatin-methylcellulose solutions was
studied at pH 3 because of a larger compatibility of the
biopolymers. Owing to the excluded volume effect, addition
of methylcellulose up to 0.2% results in an -2-fold increase in
the gelatin gelation rate (41). This is in good agreement with
the effect of addition of dextran (32), partially hydrolysed
starch and polyethylene glycols (42). Further increases in
methylcellulose concentration (above 0.2%, w/w) do not
change the gelation time of the gelatin. Enthalpy of the
junction zone rupture, calculated from the Eldridge-Ferry
equation (43), increases by adding 0.1% methylcellulose to
the gelatin gel and does not change when a larger amount of
methylcellulose is added.
Figure 10 shows the effect on rheological properties
(apparent instant Young modulus by one-axial compression
and the largest Newtonian viscosity calculated from 2 h
creep) of gelatin-methylcellulose gels of an amount of
methylcellulose added to 3 and 5% w/w gelatin solutions
(12,41). The effect of methylcellulose addition on the
166 D. V.Zasypkin, EEBraudo and V.B. Tolstoguzov

2.0 2.0

\
a
Q
-.j

Is;?
o
QJ VI
VI :J
o :J 1.2
:J u
o
QJ E
o
>-
...s:
QJ
L
0.4

5 10
._.-.-..-.---•....1'- -.-
0.5
Gelatin, %,w/w

(b)
(b)

QJ
VI
o ....>-
VI
1.0

_.--.-._-- " ---.....

:J o
<)
QJ VI 1
U
:>
a>-
QJ
~

0.1 0.3 0.5

5 10
Gelatin, %) w/w
Figure 9 Phase diagrams of the water/gelatinlmethylcellulose
system at 35°C and at pH 4.75 (a) and pH 3.0 (b). 0, phase
separation threshold; ., critical po int (40).
Methylcellulose, %,w;w
Figure 10 Effect of methylcellulose on the mechanical properties of
gelatin gels, pH 3.0, 20°C. (a) Apparent instant Young modulus of
gelatin gels. (b) Largest Newtonian viscosity. 1,3% w/w of gelatin ; 2,
5% w/w of gelatin. Ageing time of gels at 4°C was 7 days (41).

elasticity of the mixed gel is in agreement with that of
agarose for gelatin-agarose gels (Fig. 6). Comparison of the
data on the rheological properties of mixed gelatin-
methylcellulose gel (Fig. 10) with the phase diagram of the
gelatin/methylcellulose water system (Fig. 9) shows an
expansion of the incompatibility area upon gelatin gelation.
Mixed protein-polysaccharide solutions are usually more
efficient as emulsion stabilizers than individual biopolymers.
This is due to the formation of a stronger gel-like stabilizing
layer around dispersed particles (13,25,26,44-46) . Figures 11
and 12 demonstrate, as an example, the synergistic effects of
gelatin-methylcellulose mixtures as emulsion stabilizer (9).
Figure 11 shows that, unlike methylcellulose, the
emulsifying capacity of gelatin is monotonously changed .
An increase in methylcellulose solution concentration
corresponds to the transition from a dilute solution to the
semi-diluted solutions, while gelatin macromolecules are
associated (at ambient temperature) in the range of
concentrations studied. Accordingly, in the case of
methylcellulose, the transition from adsorption of isolated
macromolecules to adsorption of associated macromolecules
occurs. The similarity of the concentration dependencies of
the emulsifying capacity of gelatin and its mixture with
methylcellulose makes it possible to assume that the mixed
adsorption layers are enriched with gelatin. Figures 11 and
12 show that methylcellulose in the mixed solutions with
gelatin enhances both the emulsifying and emulsion-
stabilizing action of gelatin .
A similar, synergistic effect of mixing gelatin with
methylcellulose is also observed in the formation and
stability of foams (41). Figure 13 shows that after 12 h of
storage, the difference in volume between foams stabilized by
0.5% w/w methylcellulose and 2% w/w gelatin disappears.
The mixture (1: I) of these methylcellulose and gelatin
solutions provides both higher foaming activity and foam
stability.
 Multicomponent biopolymer gels 167

--o---u-O-o-o-o-o-o-O- 3
50 /0 _ 0 - 0 -0 - 0 - 0 - 0 - 0 - 0 - 1 120 3
? .... O ("'") o - -o-O-o_o..L.. o_
~ tl _..------------ 2
E
u

~ t // OJ
E
80 --0 ............ 0

i 3J \ I ~E
>

o
40 -0-0_ 0"-....0__ / "'" o---o_g~o_
2

rl ~ \..)
& ~o-
1

'0
~
2 6 10
Time, h
l1J
10
Figure 13 Volume of foam versus storage time, pH 3.0,25°C. 1,2%
gelatin solution; 2, 0.5% methylcellulose solution; 3, mixture of the
solutions, 1:1 (by weight) (41).
M U W
Concen l rati on . % , w/w
3
Figure II Dependence of the emulsifying capacity (E) on the
concentration of emulsifier (pH 4.75, 25°C): I , gelatin; 2, 1.6
methylcellulose; 3, gelatinlmethylcellulose I :I (by weight) blend (9).

10 0 1.2
o~

"0
u
OJ
+-
0
L.. x 0.8
0 96
a. QJ
OJ
Vl U
c 0.4 0.8 1.2 1.6 C
:J a
1.6 1.2 0.8 0.4 a.
E O.L,
Gelatin , % ,wjw a
u
Methylcellulose, 'Yo, WjW

Figure 12 Dependence of the stability of emulsions on the o 60

compositionof the emulsifier (9).
Complex gelatin/sodium alginate gels
Gels can be formed by soluble and insoluble complexes of
oppositely charged biopolymers (1--4,47,48). The solubility
and other physicochemical properties of inter-biopolymer
complexes depend on the type of unlike charged biopolymer
reagents (i.e, polyelectrolyte components), their ratio, ionic
strength and pH. Gels of soluble and insoluble complexes of
gelatin with sodium alginate (49,50) and with low esterified
pectin ate (51) were studied systematically.
The formation of inter-biopolymer complexes of gelat in
and sodium alginate occurs at pHs from 2 to 4.5 where
macro-ions bear unlike net charges.
In soluble complexes are usually precipitated in the form of
gels. The following melting and cooling of these particles
results in the formation of voluminous samples of complex
gels. Figure 14 shows the therrnornechanical behaviour of
gelatin gel (curve 1) and of the gelatin-sodium alginate
complex gel (curve 2). The melting temperature of the
20 40
Temperature) "c
Figure 14 Thermomechanical curves: I , 20% w/w of gelatin gel; 2,
complex gel of 20"10 w/wgelatinl5% w/w sodium alginate (insoluble
complexes, without ageing), pH 3.5; 3, complex gel of 5% w/w
gelatin/2.5% w/w sodium alginate (soluble complexes, gel ageing
time I day); 4, gelas in 3, gelageing time 5 days;5, complex gelof 5%
w/w gelatinl2.5% w/w sodium alginate, prepared with 8 mol/dm3
urea (soluble complexes), gelageing time 5 days.
complex gel is higher than that of the gelatin gel. Complex
gels of this kind are stable and their thermomechanical
properties do not alter for quite a long time.
Gels can also be prep ared using soluble inter-biopolymer
complexes. To this end , dry preparations, produced by dry ing
solutions of gelatin-sodium alginate complexes, are usually
used . Gels of soluble gelatin-sodium alginate complexes are
prepared by swelling and dissolving of the preliminary dried
complexes above 40°C , followed by cooling of the solution to
below 30°C. The properties of these complex gels depend
168 D. V'Zasypkin, E.E.Braudo and VB. Tolstoguzov
greatly on ageing time. The thermomechanical curves of
gelatin gel and freshly prepared complex gel are very similar
(Fig. 14, curves 1 and 2). Both kinds of gels are thermo-
reversibly melted from 30 to 40°C. Ageing (keeping for a few
hours at 10°C) results in an S-shaped thermomechanical
curve (Fig. 14, curves 3 and 4), i.e. in thermoirreversibility of
complex gels. From 30 to 40°C, gel compliance increases
sharply. An increase in ageing time of the complex gel up to
5 days leads to a decrease in compliance. Unlike gelatin gels,
complex gels based on soluble complexes can be produced in
8 mol/drn! urea. The compliance of these complex gels
containing urea does not increase in the range 30--40°C (Fig.
14, curve 5). Since 8 mol/dm! urea solution suppresses
H-bonds and hydrophobic interactions, it can be assumed
that the complex gel network is mainly formed by
electrostatic interactions of oppositely charged
polyelectrolytes during ageing of the gel (1-3, 47-51) . This
rearrangement of interrnacromolecular bonds causes the
transition from a thermoreversible to a thermo- irreversible
gel network.
Some applications of multicomponent gels
Mixtures of gel-forming agents allow the amount of gelling
agent used to be reduced and the composition-
structure-property relationship of many processed food
systems and final food products to be controlled, including
confectionery and bakery products. Both synergistic and
antagonistic effects of mixing biopolymers are of
importance for many applications (27).
The synergistic effect of single-phase mixed solutions of
biopolymers is due to the excluded volume effect of macro-
molecules. In two-phase systems, synergy results from
concentration of the continuous phase rich in stronger
gelling agent. Synergy can also result in the formation of
complex gels with novel unusual properties (4,27).
Filled, complex and mixed gels of gelatin with different
polysaccharides were used for the development of a caviar
analogue (3,9,27,52-54). Non-traditional pasta products
based on calcium alginate gels filled with starch and proteins
are described in refs 3 and 54-56. Filled anisotropic gels of
fibrous structure were used for producing meat or fish
analogues and extenders (3,10,11,16,47,48). To this end, the
processes of spinneretIess spinning and thermoplastic
extrusion were mainly used (3,10,16,57-67).
It has recently been shown (18,68) that the properties of
pressure-induced (e.g. at 450 MPa for 30 min at neutral pH)
gels of beta-lactoglobulin can be improved by the addition of
0.5-1.0% w/w xanthan, 0.1% w/w sodium alginate, 20% w/w
maltodextrin and some other polysaccharides. The excluded
volume effect, which is greatly pronounced for heat-induced
multicomponent gels, is not as significant for pressure-
induced gelation of the same systems.
Another recent application of multicomponent gels is for
producing low fat spreads and fat replacers. Fat replacers are
produced in the form of small (0.5-20 urn) dispersed
particles of protein/polysaccharide or protein/protein gels in
a highly viscous liquid or gel-like medium (13,18,69,70).
Acknowledgements
The authors are grateful to Dr Elizabeth Prior for editing the
manuscript.
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Received on January 5, 1996; accepted on May 21, 1996