Non-Covalent Interactions in Polysaccharide Systems

Marguerite Rinaudo
CERMAV-CNRS, BP53X, 38041 Grenoble cedex 9, France
Fax: 33-476547203; E-mail: marguerite.rinaudo@cermav.cnrs.fr
Received: March 8, 2006; Revised: May 14, 2006; Accepted: May 15, 2006; DOI: 10.1002/mabi.200600053
Keywords: amphiphilic polymer; hydrogen bond; hydrophobic interactions; ionic interactions; ionic selectivity; networks;
polysaccharides; physical gelation; solvation
Introduction
Knowledge of the physical chemistry of natural and modied
polysaccharides is progressing and several examples are now
available that clearly demonstrate the role of non-covalent
bonds present in aqueous solutions of polysaccharides. Be-
cause of the stereoregularityof suchpolymers, interactions are
oftencooperative andplaya role inpolymer conformationand
association in solution, even in dilute regimes.
The purpose of this paper is to give examples of poly-
saccharides that have a well-characterized chemical struc-
ture in which these interactions are clearly demonstrated.
These include Coulombic interactions (polyionion inter-
actions) and ionic selectivity, and dipolar hydrogen bonds
that affect helical structure stabilization, chain stiffness,
polymerpolymer interactions, or hydration and hydro-
phobic interactions, especially in some modied polysac-
charides (block and graft copolymer types). This paper is
based mainly on our own work in which the same methods
for purication, characterization and analysis of physical
properties have been developed.
In the following examples, only aqueous solutions are
considered: the nature of the water solvent is very important
in controlling the behavior of water-soluble polysaccharides
and its high dielectric constant results in a large contribution
of electrostatic interactions. In this respect, water plays a
crucial role: i) it causes repulsion between same charged
species to prevent the collapse and attraction between
oppositely charged entities, and ii) it has a role in the entropic
formation of hydrophobic interactions and also forms a
hydration shell around solute molecules because of its
dipolar character. Bulk water forms a highly cooperative
hydrogen-bondednetwork, inwhichthe solutewill exchange
water molecules during dissolution, which leads to a new
distribution of hydrogen bonds.
[1]
It is alsoclear that one type of non-covalent bondis usually
not independent fromothers andthe properties of the systems
result in a delicate equilibrium between the different contri-
butions, among which one dominates, depending on the
actual thermodynamic conditions.
Hydrophobic Interactions
Amphiphilic molecules dispersed in aqueous media disturb
the water structure. The hydrophilic polar part of the mole-
cules interacts with water (with hydrogen-bond formation
and ion hydration) and the hydrophobic parts interact to
form a hydrophobic domain to reduce contact with water.
From this organization, interesting new properties may
Summary: This paper describes the behavior of some poly-
saccharides withwell-knownchemical structures and inwhich
the inuence of cooperative secondary interactions play an
important role. The roles played by hydrophobic and ionic
interactions (including ionic selectivity) on polysaccharide
conformation and gelation are discussed. Electrostatic attrac-
tions are also important in the complexes formed between
surfactants and polyelectrolytes of opposite charge. Finally,
van der Waals dipolar interactions and particularly hydrogen-
bond formation are examined. The role of hydrogen bonds in
solubility, conformation, and especially the local stiffness of
polysaccharides, but also in polymerpolymer complexes
frequently obtained with polysaccharides, is developed.
Repeat unit for a number polysaccharides.
Macromol. Biosci. 2006, 6, 590610 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
590 DOI: 10.1002/mabi.200600053 Feature Article
occur. Two types of systems are examined here, based
on the chemical modication of natural polysaccharides:
methylcelluloses, which are neutral polymers, and alkyl-
ated chitosans, which are positively charged polymers
under acidic conditions. These two types of polymers have
an amphiphilic character because of the hydrophobic part
introduced by chemical modication and the hydrophilic
part composed of the polysaccharide backbone. They are
sensitive to temperature, salt concentration, and the nature
of the hydrophobic part.
Methylcelluloses
Commercial methylcelluloses are usually obtained by
methylation of semi-crystalline cellulose in the solid state;
the reaction is heterogeneous because of differences in
reactivity of the amorphous and crystalline zones. In our
work, it has been clearly demonstrated that commercial
methylcelluloses have a blockwise distribution of highly
methylatedparts of thecellulosicchains. It has beenknownfor
a long time that these block copolymers behave in a very
unusual way in aqueous solution, to showa solgel transition
at around 608C. This gelation induced by an increase in
temperature is directly related to the hydrophobic character-
istics of the polymer. The mechanism of gelation has been
examined on different commercial samples. Methylcelluloses
have been prepared in our laboratory under homogeneous
conditions and their behavior has been compared with
heterogeneous polymers.
[25]
The rst difference between the two types of methylcellu-
loses (homogeneous or heterogeneous) is that the solubility
is showntodependonthe methyl groupdistributionalongthe
chain. Homogeneous samples become water-soluble when
the degree of substitution (DSfraction of OH groups
substituted per D-glucose unit) is around 0.9, a value that is
enough to prevent the packing of the cellulosic chains in the
dried state. With heterogeneous methylcelluloses, a DS of
1.3 is needed for solubilization in relation to the existence
of non-substituted zones (Figure 1).
In Figure 2, the rheology of a commercial methylcellu-
lose from Dow Chemicals (sample A4C) at a polymer con-
centration (10 g L
1
) larger than the overlap concentration
C* as a function of temperature is given. Fromthis, it can be
observed that the situation is relatively complex: between
35 and 55 8C, a clear loose gel is formed for which G
0
>G
00
(at 1 Hz) followed by a strong increase of the G
0
modulus
above 60 8C. This second transition step occurs when the
polymer concentration is higher than the overlap concen-
tration (around 2 g L
1
for this sample) and this gel is
turbid. The gelation corresponds to a phase separation for
this low critical solution temperature polymer at a critical
temperature around 29 8C, for which the phase diagram for
this study is given in Figure 3.
[6]
The gelation is reversible
but with a large hysteresis as shown in Figure 4; this pheno-
menon has been observed not only by rheology but also by
DSC (differential scanning calorimetry) (Figure 5).
[7]
Hysteresis is often observed in physical gelation and seems
to be related to the degree of aggregation of chains and the
Marguerite Rinaudo has a Master degree (1964) and a Ph.D. (1966) from the University of Grenoble
(France) on physico-chemical properties of polyelectrolytes; her work was devoted to the synthesis and
solution properties of carboxymethylcelluloses. From that time, she continued to develop her research
work on water-soluble polymers and especially on polysaccharides (cellulose, starch and chitosan
derivatives, hyaluronan and other bacterial polysaccharides, seaweed polysaccharides); in her
laboratory she settled up a series of equipments and methodologies to investigate the specific behavior
of polysaccharides, which have to be considered as semi-rigid polymers. She started as full Professor
in 1968 at Joseph Fourier University (Grenoble-France); actually, she is Emeritus professor. During 12
years (19841996), she has been the director of a CNRS institute named Centre de Recherches sur
les Macromolecules Vegetales working on plant oligo- and polysaccharides. She received important
awards from the French government (Chevalier dans lOrdre National du Merite (1984) et dans
lOrdre de la Legion dHonneur (1998)); she became associate member of the Chilian academy of
sciences (2005) and of the Brazilian academy of sciences (1991). She published more than 400 original
papers and delivered more than 250 main lectures in international meetings. She also supervised more
than 50 PhD thesis and she is member of the editorial board of 5 international journals (from which
Biomacromolecules and Food Hydrocolloids).
Figure 1. Repeat unit for a number polysaccharides. Cellulose:
ROH, chitin: RNHCOCH
3
, chitosan: RNH
2
or
NHCOCH
3
depending on the acetylation degree (DA), alkylated
chitosan: RNH
2
or NHCOCH
3
or NH(CH
2
)
n
CH
3
depending on DA and on the degree of substitution DS.
Non-Covalent Interactions in Polysaccharide Systems 591
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stability of the aggregates formed. The same type of
behavior will also be discussed later for k-carrageenan.
The progressive aggregation of methyl groups has been
observed by
13
C NMR spectroscopy in the presence of
dimethyl sulfoxide (DMSO) as a reference; the mobility of
methyl substituents around 60 ppm is reected by the ratio
of the integral of the signal of O-Me in the C-6 position and
that of the DMSOprobe.
[6]
The results are given in Figure 6
and clearly conrm the two steps. Gelation is induced by
increasing the temperature but it also depends on the ionic
concentration (Figure 7) and on the nature of the anion and
cation that constitute the added salt. This behavior is
characteristic of hydrophobic interactions.
[5]
The sequence
for the second step of gelation, particularly as a function of
the nature of the ions and relative to the ability to induce
gelation at a given salt concentration (0.5 M) is as follows:
- for sodiumsalts: Cl

>NO
3

>without salt >I

> SCN

,
- for chloride salts: Na

>NH
4

>Li

>without
salt,
- for nitrate salts: Na

>NH
4

>without salt >Li

.
The rst ion cited is the one that induces gelation at the
lower temperature.
For homogeneous methylcelluloses the enthalpy obtained
by DSC remains very low and corresponds to loose inter-
actions between the modied cellulosic chains. From these
conclusions, the following mechanism is proposed: the clear
gel is attributed to the interaction of highly substituted zones
of the cellulosic chains; the second phase separation then
occurs, which involves the residual polymer, and the turbid
gel is formedwitha large increase of thegel modulus over C*.
Alkylated Chitosans
Chitosan is a modied natural polysaccharide obtained by
deacetylation of chitin, a constituent of the crustaceous
shells of crabs and shrimps.
[8]
Chitosan is a linear polymer
formed of partially acetylated b-(1 !4)-D-glucosamine
(Figure 1). It is soluble in acidic aqueous media by pro-
tonation of the NH
2
groups at the C-2 position; it becomes
a polycationic polymer whose charge density depends on
Figure 2. Inuence of temperature on the rheological behavior of a commercial
methylcellulose in water. Sample A4C: DS 1.7, C
p
10 g L
1
, G
0
is the
storage modulus, and G
00
the loss modulus measured at o1 Hz. Heating rate
0.5 deg. min
1
.
[5]
Figure 3. Phase diagramof commercial methylcellulose sample
A4C (DS 1.7) in water, with the cloud point curve (&) and the
sol gel transition line (&) determined by rheology. Reprinted
from ref.
[6]
, Copyright 2006, with permission of Elsevier.
592 M. Rinaudo
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
the degree of acetylation and on the degree of protonation,
depending on the concentration and nature of the acid used
for protonation.
[9,10]
It should be noted that the intrinsic pK
0
is around 6 and that chitosan is soluble in acidic media of
pH<6. In acidic conditions, chitosan, even when fully
protonated, tends to form aggregates under different condi-
tions of temperature, nature and concentration of the salt,
etc., from which it has been concluded that aggregates are
formed in solution as a result of hydrogen bonds and
hydrophobic interactions.
[11a]
This hydrophobic character
Figure 4. Rheological behavior of a commercial methylcellulose in water as a
function of successive heating and cooling. Sample A4M: DS 1.8, C
p
15 g L
1
,
G
0
is the storage modulus, and G
00
is the loss modulus measured at o1 Hz.
[5]
Figure 5. Thermogram obtained from a DSC experiment on heating and cooling of a solution of a
commercial methylcellulose. Sample A4C: DS 1.7, C
p
30 g L
1
, temperature variation rate
0.5 deg. min
1
. Reprinted from ref.
[6]
, Copyright 2006, with permission of Elsevier.
Non-Covalent Interactions in Polysaccharide Systems 593
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is not only based on the presence of N-acetyl groups but also
on the main polysaccharide backbone as mentioned below.
The reactive C-2 position allows specic reactions,
especially reductive alkylation with aldehydes. Aldehydes
of different chain lengths (from 3 to 14 carbon chains,
named C3 up to C14) have then been grafted onto the
chitosan backbone to give original properties in aqueous
solution.
[12,13]
It is recognized that the properties depend on
the degree of alkylation, the length of the alkyl chain,
temperature, pH, and on the polymer and salt concentra-
tions, as shown in Figure 8. The alkyl chains with a C12
length start to give large interchain interactions, even at
concentrations lower than the overlap concentration, to
become a physical gel depending on the ionic concentra-
tion.
[13]
The hydrophobic interactions are counter-balanced
by electrostatic repulsions that depend on the net charge of
the chitosan (i.e., on the pH) progressively screened by the
addition of salt. Alkylated chitosan with a C12 alkyl chain
(named Chit-C12) is soluble in 0.3 M acetic acid, which is a
good solvent for unsubstituted chitosanwith a characteristic
rheological solution behavior (G
00
>G
0
over a large frequ-
ency range). The complex viscosity for 6 and 10 g L
1
is
nearlyNewtonianinthe frequencyrange covered(Figure 9A).
In the presence of sodium acetate salt, even at concentrations
as lowas 0.025M, the complexviscositybecomes largelynon-
Newtonian because of loose inter-chain interactions. Gel-like
behavior (G
0
>G
00
) is obtained over 3 g L
1
as shown
earlier.
[13]
In Figure 9B, solution behavior with G
0
<G
00
is
obtained in the absence of salt but turns to a gel-like behavior
(G
0
>G
00
) when some electrostatic interactions are screened
by the addition of salt (Table 1).
The role of pH and temperature is shown in Figure 10.
When the pH increases, the net charge of the polymer
Figure 6. Relative intensity of the signals obtained for the OMe
substituent at the C-6 position by
13
C NMR spectroscopy as a
function of temperature for a methylcellulose in D
2
O.
Sample A4C: DS 1.7, C
p
44.6 g L
1
. Reprinted from ref.
[6]
,
Copyright 2006, with permission of Elsevier.
Figure 7. Inuence of salt concentration on the rheological behavior of a commercial
sample as a function of temperature in aqueous medium. Sample A4M: C
p
10 g L
1
, G
0
is the elastic modulus, and G
00
is the viscous modulus measured at o1 Hz.
[5]
594 M. Rinaudo
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
(related to the degree of protonation, a) decreases, hydro-
phobic attractions dominate, and gel-like behavior is
observed with a large increase in the apparent viscosity
followed by a phase separation under a critical value of a. At
the same time, it is shown that raising the temperature
increases the observed maximum viscosity. This behavior
provides evidence of the existing balance between electro-
static repulsion and hydrophobic attraction, with results
summarized in Figure 11.
[14]
The bulk properties of these associating polymers are
very important for use as thickeners with an important non-
Newtonian character, because of the loose and reversible
inter-chain interactions. This mechanism is well demon-
strated when b-cyclodextrins (CDs) are added to C12
alkylated chitosan. The very high viscosity is reduced by the
progressive addition of CD because of alkyl chain/CD
complex formation, which leads to chain separation and
viscosities typical of derivatives with shorter alkyl chains
(Figure 12).
[13]
The micelle-like junctions form hydro-
phobic domains in a semi-dilute regime as shown by
modication of the uorescence spectrum of pyrene
added as a probe in solution. The uorescence spectrum
of pyrene is modied in relation to the polarity of the
environment. It has been shown that these hydrophobic
domains are able to adsorb hydrophobic substances such as
taxol or B
2
vitamin and behave as a reservoir for controlled
drug release.
[15,16]
Another important characteristic of amphiphilic poly-
mers is their interfacial activity. It has been shown that
alkylated chitosan moderately decreases the air/water and
oil/water interfacial tension but the effect increases with
alkyl chain length, as shown previously.
[13,1720]
In addi-
tion, it has been shown that the mechanical properties of the
interfacial lm formed are completely changed compared
to those of simple surfactants.
[21]
Acooperative interaction, normal with polyelectrolytes,
is obtained between chitosan and oppositely charged
surfactants, which leads to a surfactant-polyelectrolyte
electrostatic complex (SPEC).
[1921]
The interfacial lmis
a nanostructured lm stabilized by strongly interacting
surfactants and is able to encapsulate molecules.
[20,22,23]
Capsules have been obtained from a chitosan gel layer
cross-linked by charged surfactant micelles and used to
encapsulate enzymes as mentioned by Babak et al.
[20,22,23]
The difference between a SPEC and alkylated chitosan
having the same density of alkyl chains with the same
length per polymeric chain is very important. In fact, in the
covalently alkylated derivative, a random distribution
of the alkyl chain substituents is obtained under the
experimental conditions adopted,
[12]
while in a SPEC
there is a cooperative interaction between alkyl chains of
the surfactants, which involves few polymeric chains as
soon as the distance between the ionic sites along the
polyelectrolyte is in the range of 0.5 nm, as claimed by
Osada and co-workers.
[24]
A critical aggregation concen-
tration (c.a.c) has been found from surface tension
measurements to be around 100 times smaller than the
critical micelle concentration (c.m.c) of the surfactant
alone.
[19]
The role of sulphated N-acyl chitosan (S-Cn-
Chitosan) on a lipidic membrane has been compared to that
of sodium dodecyl sulfate (SDS). SDS dissociates the
membrane while the polymer increases the rigidity of the
membrane, which suggests low toxicity on bioorganisms.
In solution when the alkyl chain in S-Cn-chitosan is longer
than n 10, these polymers formmore stable micelles than
the micelles formed with the same alkyl chain surfac-
tant.
[25]
An extension of this interaction comes in the eld
of food application. It involves specic interactions with
phospholipids and bile acids.
[26]
Ionic Interactions
A solution of charged polysaccharides has an original
set of properties as a result of electrostatic interactions
that involve all the ionic species in solution. The rst
type of interaction involves attraction between polyions
and oppositely charged ions such as counter-ions, ionic
surfactants, or polymers. The second is repulsion between
species with the same charge such as long-range electro-
static interactions between chains that cause unusual
viscosity behavior, which reaches a maximum when the
reduced viscosity is plotted as a function of polymer con-
centration at lowionic concentration or in light scattering or
neutron scattering under the same conditions.
[2729]
This
second type of electrostatic interaction will not be covered
in this paper.
Figure 8. Viscosity of alkylated chitosan in 0.3 M AcOH/0.1 M
AcONa at zero shear rate as a function of polymer concentration at
25 8C. (&) Chitosan, (*) Chit-C6, (~) Chit-C8, (^) Chit-
C10, and (
&
) Chit-C12. Reprinted with permission from
ref.
[13]
, Copyright 2006, American Chemical Society.
Non-Covalent Interactions in Polysaccharide Systems 595
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Counter-Ion Interaction and Ionic Selectivity
Electrostatic interactions between a charged polymer and
counter-ions in the absence of external salt is governed by
the electrostatic potential around the polyelectrolyte as
exposed in the Katchalsky or Manning theories.
[30,31]
The
linear charge density imposes the activity coefcient of the
counter-ions as well as the limit over which ionic selectivity
appears related to the condensation limit, as proposed by
Manning. These theories have been compared in a previous
Figure 9. Role of salt on the rheological behavior of alkylated chitosan (Chit-C12) as a function of frequency at 10 8C:
A) complex viscosity in 0.3 M AcOH (^ 6 and & 10 g L
1
) and in 0.3 M AcOH/0.025 M AcONa (} 6 and &
10 g L
1
),; B) storage (G
0
&) and loss (G
00
&) moduli in 0.3 M AcOHand G
0
(*) and G
00
(*) in 0.3 M AcOH/0.025 M
AcONa. Polymer concentration: 10 g L
1
.
Table 1. Inuence of salt concentration on the rheology of alkyl
chitosan. (Concentration 10 g L
1
; o1 Hz.)
Solvent G
0
G
00
jZ*j
Pa Pa Pa s
0.3 M AcOH 1.75 3.73 0.66
0.3 M AcOH/0.025 M AcONa 6.75 5.79 1.42
596 M. Rinaudo
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
paper on solution properties (activity coefcients) of a
series of carboxymethylcelluloses with different degrees of
substitution.
[32]
It is known that electrostatic ionpolyion
interactions become independent of the chain length as
soon as the degree of polymerization is larger than a value of
approximately 20. The different main electrostatic proper-
ties of some polysaccharides have recently been described,
based on some of our results.
[29]
From a general point view,
and based on these theories, it is possible to predict with
reasonable condence the activity coefcient of counter-
ions in the absence of external salt in puried ionic poly-
saccharides as soon as the structure is known (nature of
the chain, degree of substitution, actual conformation and
distribution of the ionic sites along the polymeric chains).
With regard to these systems, it has also been shown that for
a charge parameter larger than 1, ionic selectivity occurs, as
demonstrated by different methods but the most clearly by
ultrasound absorption.
[33,34]
This behavior has clearly been
demonstrated for carboxymethylcelluloses obtained by
homogeneous substitution on the laboratory scale, where
the ionic selectivity is found to be:
Li

> Na

> K

> Cs

> TEA

with Li

forming more ion pairs and tetraethyl ammonium,

a large hydrophobic ion used as reference, being inactive.
Data concerning different aspects of counter-ion interac-
tions are described below.
For gellan and k-carrageenan, particularly in salt free
solutions, it has beendemonstratedthat over a critical polymer
concentration (which alsomeans a critical ionic concentration
Cs*), at a given temperature, a double helix forms that is
stabilized by hydrogen bonds. When the temperature is
increased, destabilization of the hydrogen bonds induces a
transition to a more exible coiled conformation as observed
by NMR spectroscopy, optical rotation, or viscometry. The
example obtained for gellan is giveninFigure 13, in which the
reduced viscosity decreases sharply around 308C. This
phenomenon will be discussed later but it must be pointed
out that the charge density in the helical conformation is
double that of the coiled conformation (Table 2). The electro-
static interaction with counter-ions then increases during the
coil-helix transition as demonstrated by a decrease in the
activity counter-ion.
[35,36]
An example is given in Figure 14
for k-carrageenan, where counter-ion activity is plotted as a
function of the polymer concentration (corresponding to an
Figure 10. Reduced viscosity of alkylated chitosan (Chit-C12)
under the chlorhydrate formin water as a function of the degree of
protonation (a) at different temperatures. Polymer concentration:
0.9 g L
1
, DS0.04. Reprinted with permission from ref.
[14]
,
Copyright 2006, Wiley-VCH Publishers, Inc.
Figure 11. Phase diagramfor Chit-C12 deduced fromthe results
of Figure 10. Reprinted with permission from ref.
[14]
, Copyright
2006, Wiley-VCH Publishers Inc.
Figure 12. Inuence of the progressive addition of b-cyclo-
dextrin on the viscosity of chitosan and alkylated chitosan
solutions at 2 g L
1
and 25 8C. (^) Chitosan, (&) Chit-C6,
(~) Chit-C8, (*) Chit-C10, and (^) Chit-C12. The ratio [CD]/
[Cx] is the molar ratio of CD and of alkyl chains with different
chain lengths (x). Reprinted with permission from ref.
[13]
,
Copyright 2006, American Chemical Society.
Non-Covalent Interactions in Polysaccharide Systems 597
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increase in the ionic concentration and a screening effect of
the repulsions between ionic sites on the chains). It is shown
that the activity coefcient for Na

remains nearly unchanged

with ionic concentration and temperature in the domain
covered, the value of which corresponds to the prediction for a
single chain potential. At a lower temperature (15 8C) for K

counter-ions, a transition is observed that reduces the activity

coefcient by nearly half the initial value, which corresponds
to the formation of a dimer. The temperature for the confor-
mational transition T
m
is directly related to the total ionic
concentration, as representedbya phase diagramproposedfor
k-carrageenan and gellan. A linear plot is obtained when the
temperature of conformational change values (T
m
) obtained
from the cooling curves by optical rotation, conductivity, or
viscosity, are represented for different polymers and external
salt concentrations (Figure 15).
[3537]
These phase diagrams
separate coil fromdouble helical conformations as a function
of ionic concentration and temperature. It should be recalled
that, from this graph (see Figure 15a) over a critical ionic
concentration (Cs**) (which may be larger than Cs*) the
double helices associate to forma gel; these critical values for
gellan and k-carrageenan have been discussed previously.
[36b]
An important difference between these two polymers is that
no ionic selectivity between monovalent counter-ions is
observed for gellan, which has a lower charge parameter.
With gellan, divalent counter-ions favor the helix transi-
tionas seeninFigure 15b, but withk-carrageenan, onlysmall
differences between divalent counter-ions are observed.
[35]
The phase diagram lines for divalent counter-ions (Ba, Ca,
Sr, Mg, Zn, and Co) are very close together, parallel to those
of Na

and K

counter-ions, and located just between their

two lines.
Figure 13. Reduced viscosity as a function of temperature for gellan under sodium and
potassiumsalt forms. Concentration 0.3 g L
1
at 0.025 Msalt concentration. Reproduced
with permission of John Wiley & Sons from ref.
[36a]
Table 2. Activity coefcients of monovalent counter-ions and molecular weights of polysaccharides in relation to their conformation.
Polysaccharide l charge
parameter
g activity
coefcient
calculated
g activity
coefcient
experimental
M
w
Ref.
K-carrageenan coil 0.68 0.72 0.37 3.4 10
5
[35]
15 8C, K form (helix);
C
p
>10
2
monomol L
1
helix 1.65 0.37
K-carrageenan 0.72 1.76 10
5
[35]
45 8C, K form (coil);
C
p
<10
3
monomol L
1
Gellan Na form coil 0.37 0.83 0.67 4.9 10
5
[36a]
18 8C (helix) helix 0.75 0.69
Gellan Na form 0.76 2.5 10
5
[36a]
26 8C (coil)
598 M. Rinaudo
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
The nature of the counter-ions and ionic selectivity is
an interesting effect based on specic interactions that
involve hydration of the ionic species. Spectroscopic techni-
ques have been used to point out the existence of ionic
selectivity that occurs when the charge parameter is larger
than a value of 1.
[34]
By ultrasound absorption, which is a
good technique to investigate ion pair formation based on
perturbation of the solvation shells, it has been demonstrated
for k-carrageenan that even if the charge parameter is lower
than 1, ionic selectivity in the coil conformation can be
demonstrated with K

forming more ion pairs then Na

.
[38]
This also shows that the same selectivity plays a role to
induce the coilhelix transition and control the stability of
the double helix and that of the gel formed. The formation of
more ion pairs further reduces the net charge of the polymer,
which favors the association of two chains to form a double
helix and then association for gel formation. The ionic
selectivity observed for gellan in solution plays a role in the
ability to associate, as shown by the increase in viscosity
under the K

form (Figure 13), but also on the mechanical

properties of the gel formed (Figure 16).
Anion selectivity also exists in carrageenan as demon-
strated by NMR spectroscopy.
[39]
It has also been shown
that I

has a specic interaction with k-carrageenan, to

stabilize the helical conformation and prevent the aggrega-
tion of double helices and gelation.
[40,41]
The mechanismof
interaction between anions and this negatively charged
polymer is still not clear but the xation of I

certainly
increases the charge density of the polymer and increases
the electrostatic repulsions between the double helices.
The same conclusions and the same model for gelation
given for k-carrageenan are also valid for gellan. The only
difference is that, because of the lower charge density of the
coiled conformation, no ionic selectivity is observed with
gellan, thoughit does appear when the double helix is formed
(Figure 15b).
[36,37]
The ionic selectivity then appears for the
double helix conformation with the ability to induce gelation
withthe followingsequence amongmonovalent counterions:
K>Na >Li >TMA.
At the same time it has been shown that the physical
properties (E, the elastic modulus and the stress at break F
m
determined in compression experiments) of the gel are
controlled by the nature of the counter-ions (Table 3).
[37]
Ionic selectivity is also recognized in alginates and pectins
in the presence of divalent counter-ions. On a poly(1,4-D-
galacturonic acid) (fully demethylated pectin), the transport
coefcient determined from conductivity (not far from the
activity coefcient) indicates a specic behavior for Mg
counter-ions compared with the other divalent counter-ions
(Table 4).
[42]
From these results it was concluded that under
Na and Mg salt forms, the general behavior of single chain
polyelectrolytes is conrmed, while with Ca, Ba, and Sr, a
double chain structure is stabilized.
[42,43]
These results are in
agreement with previous data by Kohn et al. and the egg-box
model mechanismrst suggested for alginate by Rees andco-
workers.
[44,45]
In addition, an interesting relation is madewith
the ability to form a gel: Na and Mg forms are water soluble
while the other forms are not, while Ca, Ba, and Sr are known
to induce gelation with an ionic selectivity corresponding to
Ba >Sr >Ca for the ability to form a gel (Figure 17).
[46,47]
The same sequence of selectivity and the mechanism are
involved for gelation of alginates in the presence of divalent
counterions.
Pectins with different degrees of esterication (DE) have
been prepared; demethylation is performed under alkaline
conditions (giving a random distribution of the carboxylic
Figure 14. Activity coefcient of monovalent counterions (K

and Na

) as a function of
polymer concentration (C
p
) at 15 and 35 8C. Reproduced with permission from ref.
[35a]
,
Copyright 2006, American Chemical Society.
Non-Covalent Interactions in Polysaccharide Systems 599
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
sites) and using an enzyme (giving a blockwise distribution of
free carboxylic groups). The charge parameter of the polymers
that have the same intermediate degree of methylation
(DE30) but different distributions does not behave in the
same way in relation to electrostatic interactions. Even if the
average charge parameter is the same, the local electrostatic
elds are not; the blockwise distribution (at least eight
carboxylic groups per block) has the same charge parameter as
the fully demethylated polymer (but not the average value
based on the degree of esterication). The gelation ability then
depends on the charge distribution as shown in Figure 18.
[46]
It
also plays a role in cell wall structure.
[48]
Figure 15. Phase diagram giving the inverse of the temperature that corresponds to the
conformational change T
m
as a function of total salt concentration C
T
: a) for k-carrageenan
under sodium and potassium salt forms; Cs** is the critical salt concentration above which
gelation occurs. Reprinted with permission from ref.
[35a]
, Copyright 2006, American
Chemical Society. b) For gellan under monovalent ( Na, o K, ~ TMA, and ~ Li) and
divalent counterions (& Ca and & Mg). Reprinted from ref.
[36b]
, Copyright 2006, with
permission from Elsevier.
600 M. Rinaudo
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Polyelectrolyte Complex
The most important systems developed recently are the
electrostatic complexes that are formed when two polymers
with opposite charges are mixed. With alginate or HA in
particular, a pH-dependent complex is formed in the
presence of chitosan, whose stability depends on ionic
concentration and pH. Complex formation has been investi-
gated in dilute solution by pH-potentiometry and conductiv-
ity to determine the fraction of ion pairs (COO

/NH
3

)
formed, depending on the experimental conditions.
[49,50]
Interactions between chitosan and alginate give an electro-
static complex that has been mostly used so far for biological
applications. The main applications of these electrostatic
complexes cited are antithrombogenic materials, controlled
release materials, encapsulation of drugs, immobilization of
enzymes and cells, and gene carriers. One aspect of these
complexes now under development is the preparation of
layer-by-layer polyelectrolyte capsules or lms based on
charged biocompatible polysaccharides or chitosan/syn-
thetic polyelectrolytes.
[51,52]
Porous gels (sponges) can be
prepared by forming a calcium alginate gel stabilized by
complexation with galactosylated chitosan (a water-soluble
derivative).
[53]
Another type of complexed bead can be
obtained by the drop-wise addition of Na-alginate into a
chitosan-CaCl
2
solution; the swelling of the beads being
different from that of Ca-alginate beads with a maximum
swelling at pH 9.
[54]
Dipolar Interactions and Hydrogen-Bond
Network Formation
Hydrogen bonds are loose dipolar interactions that involve
hydrogen atoms interacting with highly electronegative
atoms such as oxygen or nitrogen. Hydrogen-bond inter-
action analysis is very important to understand the
physicochemical properties of polysaccharides in aqueous
solution. These polymers are very rich in OH groups or
C O. At low temperatures, hydrogen bonds can be formed
to give rise to intrachain or interchain networks. Interaction
with the solvent (water) is also favored in relation to
solvation and dissolution. Usually, the existence of ionic
sites along the chains prevents interchain interactions,
which exist for neutral polysaccharides. The situation of
galactomannans is clear in this respect and has been
discussed previously;
[55]
in this case loose interactions
exist, which lead to loose gel-like behavior in solution.
Solubility
Froma general point of view, the greater the interaction of a
solute with solvent molecules, the greater the solubility. In
Table 3. Physical properties of gellan in the gel form under
different ionic forms. Gellan concentration C
p
10 g L
1
; salt
concentration [XCl] 0.1 M. Reprinted from ref.
[36b]
, Copyright
2006, with permission from Elsevier.
X E (10
4
) F
m
Pa N
Li 0.44
Na 11.9 13.3
K 31.4 23.1
Figure 16. Elastic modulus E for puried gellan under potas-
sium () and sodium (*) forms as a function of polymer
concentration in salt excess. Salt concentration 0.1 M, temperature
25 8C. Reprinted from ref.
[36b]
, Copyright 2006, with permission
from Elsevier.
Table 4. Transport coefcient obtained by conductivity for
different counterions in a poly-D-galacturonate solution.
Reprinted from ref.
[42]
, with permission of John Wiley & Sons.
Counter-ion Transport coefcient of
polygalacturonate
a)
Na 0.62
Ba 0.095
Ca 0.11
Sr 0.11
Mg 0.28
a)
l charge parameter for a single chain 1.58 and for a double
chain 3.16.
Non-Covalent Interactions in Polysaccharide Systems 601
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
fact, solubilization of polysaccharides is a difcult problem
because of the existence of a cooperative network formed
by interchain hydrogen bonds. The difculty in solubilizing
such a system in water comes from too high a temperature
and total dehydration when puried polysaccharides are
rst isolated making OH groups no longer accessible to
interact with water molecules. The existence of charged
groups such as SO
3

or COO

favors the dissociation of

Figure 17. Inuence of the progressive addition of divalent counter-ions on the
critical gel point for polygalacturonic sodium salt form followed by light scattering.
Reproduced with permission from ref.
[46]
, Copyright Society of Chemical Industry.
Permission granted by John Wiley & Sons Ltd on behalf of the SCI.
Figure 18. Inuence of the distribution of carboxylic groups along the polygalactur-
onic chain for DE 30 pectins obtained by enzyme (PE: blockwise distribution) and by
alkaline treatment (PH: random distribution) on gelation in the presence of calcium
counter-ions. Reproduced with permission from ref.
[46]
, Copyright Society of
Chemical Industry. Permission granted by John Wiley &Sons Ltd on behalf of the SCI.
602 M. Rinaudo
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
aggregates by electrostatic repulsion and increases the
ability to x water because of the presence of the ionic sites
and their counter-ions.
[56]
The progressive hydration of HA
has recently been studied by infrared spectroscopy.
[5759]
This shows why heterogeneous methylcellulose with a
DS 1 is not water solublewhile homogeneous samples are
soluble, the dimension of the methyl substituents regularly
dispersed along the chain prevent the packing of poly-
hydroxylated molecules.
In the majority of oligo- and polysaccharides, the hydro-
philic character as a result of the OH groups is counter-
balanced by the hydrophobic contribution of the CH entity
of the glycosidic cycle and the ether-like O-4 and O-5 oxygen
atoms.
[60]
Such hydrogen-bond formation and the additional
hydrophobic character of polysaccharides have been demon-
strated for chitosan under acidic conditions.
[11a]
With
hyaluronan, the self association of segments in 0.15 M NaCl
has also been discussed.
[11b]
These associations are related to
the local stiffness of the polysaccharides which promotes
cooperative interactions.
The interaction of water molecules with a model molecule,
maltose, has been approached by molecular modeling.
[61]
The locationof water molecules has been described (albeit by
a dynamic process) and the results have been used to predict,
with good agreement, the hydration of amorphous amylose
(on average, 3.8 water molecules are rmly interacting with
one D-glucose moiety). Experiments on amorphous amylose
conrm the prediction and have been extended to acetylated
starch in relation to the inuence of the relative humidity on
the mechanical properties of lms.
[62,63]
Hydrogen bonds play an important role in solubility, as
has recently been investigated for cyclodextrins.
[64]
The
role of hydrogen bonds has been displayed by comparing
their behavior in H
2
O and in D
2
O. In D
2
O, stronger
hydrogen bonds are formed than in H
2
O and the solubility
of CDdecreased in D
2
O. We have also shown that when the
helical conformation of polysaccharides is stabilized
by intrachain hydrogen bonds, the conformation is more
stable in D
2
O, as has been found with xanthan and other
stereoregular polysaccharides. The example obtained for
gellan in H
2
O and D
2
O is shown in Figure 19, where the
optical rotation is plotted as a function of temperature,
[36]
and the double helixcoil transition temperature increases
by around 6 8C in D
2
O under the experimental conditions
used. Another interesting behavior in relation to hydrogen-
bond formation has been obtained regarding hyaluronan
viscosity (Figure 20).
[65]
When the pH decreases, the net
charge of the polymer decreases as well as the electro-
static repulsions between polysaccharide chains, which
favors the stabilization of aggregates to give a gel-like
behavior. In this situation, pH2.4 may also represent a
pseudo isoelectric point as a result of an equilibrium
between a few anionic groups from the uronic unit and the
amino groups from the N-acetyl-D-glucosamine unit (see
Figure 21).
Conformation and Stiffness
Hydrogen bonds play an important role in the local
conformation of stereoregular polysaccharides as many of
them adopt a helical conformation in the solid state, as has
been established for gellan,
[66]
k-carrageenan,
[67]
hyaluro-
nan,
[68]
chitin,
[69]
and xanthan.
[70]
In aqueous solution, under
given thermodynamic conditions, they adopt an ordered
conformation assumed to be helical, as displayed by circular
dichroism, optical rotation, or NMR spectroscopy, while the
conformational transition is induced by increasing the
temperature or changing the ionic concentration.
[3537,71,72]
The transition results from a decrease in the attraction by
hydrogen bonds when the temperature increases and the
effect of electrostatic repulsions between ionic groups on the
chain.
The intrachain hydrogen bonds in such an ordered
conformation are very important because they control the
local stiffness of the polymer, reected by its intrinsic
persistence length (L
p
), as seen, for example, for hyaluronan
in Figure 21. Some L
p
values are given in Table 5. In relation
to this rigidity, the worm-like chain model was introduced a
few years ago to analyze the behavior of polysaccharides,
the local stiffness of the polymer being characterized by an
intrinsic persistence length L
p
.
[73,74]
For a charged polymer,
it is necessary to introduce an electrostatic contribution L
e
(the total persistence length being L
t
L
p
L
e
at a given
ionic concentration) following the Odijk development.
[75]
The local stiffness of the polysaccharides allows the high
viscosity obtained to be interpreted for a given molecular
weight compared to a exible polymer. The L
p
value,
obtained in a large excess of salt, is characteristic of the
local structure of the polysaccharides and can be predicted
from molecular modeling, as recently shown.
[7678]
The experimental values of intrinsic viscosity and radius
of gyration can, therefore, be interpreted as a function of the
molecular weights of the polysaccharides. Theoretically,
the persistence length L
p
of a chain and the molecular
weight directly control the radius of gyration R
g
and the
intrinsic viscosity [Z] on the basis of the worm-like chain
model under y conditions, following the Benoit Doty
equation:
[79]
R
2
g
L L
p
=3 L
2
p
2L
3
p
=L 2L
4
p
=L
2
1 expL=L
p

1
in which L is the contour length calculated from the
chemical structure and the molecular weight (M). For high
molecular weights, one can limit the analysis to the rst
right-hand term of Equation (1). The intrinsic viscosity is
given by the formula proposed by Yamakawa and Fujii:
[80]
Z FM; L
p
; dM
L
=2L
p

3=2
M
1=2
2
with d, the hydrodynamic diameter, and M
L
, the mass per
unit length of the chain, which is known from the chemical
structure.
Non-Covalent Interactions in Polysaccharide Systems 603
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Experimentally, the intrinsic viscosity [Z] (expressed in
mL g
1
) can be obtained directly from steric exclusion
chromatography (equipped with a refractometer, viscom-
eter, and light scattering detectors on line) at the same time
as the molecular weight but, more usually, it is determined
in a separate experiment. It is obtained by extrapolation to
zero polymer concentration of the reduced viscosity (Z
red
)
of the polymeric solution plotted as a function of the
polymer concentration (C in g mL
1
) in accordance with
the Huggins relation:
Z
red
Z Z
0
=Z
0
C Z k
0
Z
2
C 3
in which Z is the viscosity of the polymer solution, Z
0
the
viscosity of the solvent, and k
0
the Huggins constant.
Figure 19. Determination of the conformational transition on gellan by optical rotation as
a function of temperature in H
2
O () T
m
24 8C and in D
2
O (*) T
m
31 8C. Polymer
concentration 6 g L
1
. Reprinted with permission of John Wiley & Sons from ref.
[36a]
Figure 20. Complex viscosity of hyaluronan as a function of the pH at 25 8C.
Polymer concentration 10 g L
1
, frequency 1 rad s
1
. Reprinted with permis-
sion from ref.
[65]
, Copyright 2006, American Chemical Society.
604 M. Rinaudo
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
In dilute and semi-dilute solutions, the most important
characteristic for applications of water-soluble polysac-
charides is the thickening character of the solution, which
depends on the intrinsic viscosity, molecular weight, and
concentration of the polymer at a given temperature and in a
given solvent.
In this case, the viscosity of a polymer solution at zero
shear rate is directly described by the following equation:
[81]
Z
sp
CZ1 k
1
CZ k
2
CZ
2
k
3
CZ
3

4
in which k
1
0.4, k
2
k
1
/2!; k
3
k
1
/3!. This relation is
very important, as it shows that the overlap parameter
(C [Z]) controls the viscosity of the solution at zero shear
rate and allows this behavior to be predicted. For a given
polymer, under given thermodynamic conditions, all the
viscosity values for different polymer concentrations and
molecular weights appear on the same curve when plotted
as a function of C [Z]. The k
1
value represents the Huggins
constant and for many perfectly water-soluble polysac-
charides it equals 0.4. All deviations from this reference
curve indicate interchain interaction or aggregation, such as
that observed with galactomannan.
[55]
The intrinsic viscosity is related to the molecular weight
of the polymer, following the empirical MarkHouwink
relationship:
Z KM
a
v
5
in which M
v
is the viscometric-average molecular weight,
and Kand a are two parameters that depend on the polymer,
solvent, and temperature. The important thing is that poly-
saccharides usually deviate from the behavior of synthetic
polymers, which are exible, and it is demonstrated that
even under y conditions, for polysaccharides, the a
exponent of the Mark-Houwink relationship is never equal
Figure 21. Hyaluronan: A) repeat unit, B) local conformation of a segment of hyaluronan,
representation of intrachain hydrogen bonds.
Table 5. Intrinsic persistence length for different polysaccharides
obtained by molecular modeling and experiments at 25 8C.
Polysaccharide L
p
Ref.
nm
HA
a)
8 [82]
HA
b)
7.5 [77]
Xanthan Coil
a)
5 [73b]
Xanthan Helix
a)
40 [73b]
Gellan Coil
a)
5 [36]
Gellan Helix
a)
35 [36]
Chitin
b)
12.5 [78a]
Chitosan DA0
a)
10 [78b]
Chitosan DA0
b)
9 [78a]
Mannan
b)
14.5 [76b]
Galactomannan M/G1:1
b)
9.6 [76b]
Galactomannan M/G1:1
a)
9.3 [76a]
Alginate M/G0.28
a)
9 [73b]
M/G1.92
a)
4 [73b]
Succinoglycan Helix
a)
35 [73b]
Succinoglycan Coil
a)
5 [73b]
a)
Determined experimentally.
b)
Determined by molecular modeling.
Non-Covalent Interactions in Polysaccharide Systems 605
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
to 0.5, excluding, for example, the direct use of the Flory-
Fox relation.
The persistence length of polysaccharides depends mainly
onthe intrachainhydrogenbonds. For example, it reduces the
mobility of the proton as determined by NMR spectroscopy
and the signals obtained depend directly on the temperature,
as has been described before and conrmed by molecular
modeling for hyaluronan and chitosan.
[82a,82c]
A stronger
effect is obtained when the helical conformation is stabi-
lized; the mobility is reduced so much in a solution of
polysaccharides in the helical conformation that all the
characteristic broad signals disappear. When the temperature
Figure 23. Storage modulus (&G
0
) and loss modulus (&G
0
) for a LBG/Xanthan (50/50) mixture in water
as a function of temperature on heating. Frequency 1 Hz.
Figure 22. DSC thermograms obtained on heating (0.2 deg. min
1
) for xanthan/
galactomannan mixture (4:2 g L
1
): a) in water, b) in 5 10
3
M NaCl, and c) in
10
2
M NaCl. T
gel
is the temperature for melting of the mixed gel and T
m
is the
temperature for the conformational transition of xanthan. Reproduced with
permission from ref.
[86]
, Copyright 2006, Wiley-VCH Publishers Inc.
606 M. Rinaudo
Macromol. Biosci. 2006, 6, 590610 www.mbs-journal.de 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
increases under given conditions (salt concentration, nature
of counter-ions, and polymer concentration), a conforma-
tional transition is observed that corresponds to the appear-
ance of the characteristic signals for protons, such as the
results obtained for succinoglycan and gellan.
[36b,71]
This
helix-coil conformational transition is related to the
equilibrium between electrostatic repulsions and hydrogen-
bond attractions.
Polymer/Polymer Complex
Usually, when two different polymers in solution are mixed,
a phase separation occurs, while in many polysaccharide
systems in aqueous solutions cooperative interactions are
established, which increases the viscosity up to physical
gelation. This phenomenon is well known when xanthan is
mixed with galactomannans or glucomannans.
[8388]
When
deacetylated xanthan and galactomannan are mixed, it has
been shown that two types of gel exist:
- Alowtemperature gel that melts at around 25 8C, which
involves disordered xanthan (coil conformation) and
galactomannan with different mannose/galactose ratios at
a lowionic concentration. These forman ordered complex
stabilized by the hydrogen bond network, which is demon-
strated by circular dichroism and differential scanning
calorimetry (Figure 22).
[8387]
- A second type of gel formed in water or in the presence
of salts especially with locust bean gum (LBG) galacto-
mannan with a relatively high mannose/galactose ratio,
with a melting point of around 60 8Cas shown in Figure 23.
The melting temperature is estimated from the crossing
point of G
0
and G
00
(Table 6).
[88]
The interaction is clearly
demonstrated in Figure 24, where the storage modulus is
plotted for the galactomannan and xanthan solutions
(10 g L
1
in water) as a function of frequency. From these
data, it can be seen that a loose interaction exists for the
xanthan/guar mixture as the modulus is larger than that of
the corresponding additivity; a larger increase of the
modulus is obtained for the mixture with LBG. In
Figure 25, the change in behavior for LBG, from a
viscoelastic solution to a gel (G
0
>G
00
in all the frequency
ranges covered) is shown. The largest modulus is obtained
after rst heating above 70 8C and then cooling. It also
shows that the modulus decreases in the presence of
external salt but that the melting temperature increases
(data not shown). The modeling of the interactions between
the two polymers (xanthan/galactomannan) has been
investigated by Chandrasekaran and Radha
[89]
and different
hydrogen-bonding interactions may involve side-chain/
side-chain interactions, main-chain/main-chain interac-
tions, etc.
Figure 24. Storage modulus as a function of the frequency for the different systems tested at 25 8C and
10 g L
1
in water.
Table 6. Rheology of galactomannans, xanthan, and their 50:50
mixtures. (Concentration 10 g L
1
; solvent H
2
O; o1 Hz.)
Sample G
0
G
00
j j T
F
(G
0
G
00
)
Pa Pa Pa s 8C
Guar M/G1.7 5.05 6.39 1.30
LBG M/G2.44 4.46 9.79 1.72
Xanthan 18.06 5.52 3.02
Xanthan/LBG 148.6 11.33 23.72 56.5
Xanthan/Guar 16.13 7.87 2.87 50.9
Non-Covalent Interactions in Polysaccharide Systems 607
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Conclusion
This paper covers the secondary interactions that exist in
aqueous solution for a few representative polysaccharides.
This includes the role of hydrophobic interactions in amphi-
philic derivatives such as heterogeneous methylcelluloses,
whichare knowntoformgels whenthe temperature increases.
The mechanism of gelation and the inuence of the different
thermodynamic parameters are discussed. The role of
heterogeneity of methyl group substitution (blockwise distri-
bution) is clearly demonstrated. Grafted alkylated chitosans
have original bulk properties and give rise to large increases in
solution viscosity up to gelation, depending on the solvent
composition as a result of the aggregation of grafted alkyl
chains that form hydrophobic domains which act as points of
contact. These derivatives also have interesting interfacial
properties with the formation of stiff lms able to stabilize
emulsions.
Ionic interactions are also of great interest. Charged
polysaccharides are characterized by an electrostatic poten-
tial, which imposes the counter-ion distribution depending
on the conformation. The activity coefcient of counter-
ions may be predicted when conrmation and structure of
the linear chain are known. For higher charge densities,
ionic selectivity is shown to play an important role in some
stereoregular polysaccharides. Ionic selectivity based on
ion pair formation controls the stability of the conforma-
tion, the more ion pairs that are formed, the more stable is
the helical conformation that results in a balance between
electrostatic repulsions and attractive hydrogen bonds. As a
consequence of this, ionic selectivity is related to the ability
of gelation and the mechanical properties of the gels.
Finally, the importance of the hydrogen bond in these
systems is discussed. They play a role in the local stiffness
and persistence length and also rheological properties in
solution. The cooperative hydrogen bonds allow the
stabilization of the helical conformation whose stability
depends on the temperature, salt concentration, and nature
of the counter-ions. Hydrogen bonds are alsoinvolvedin the
hydration and dissolution of polysaccharides. Interchain
hydrogen bonds allowthe formation of original cooperative
polymer A/polymer B complexes, against the usual rule of
polymer incompatibility.
In all these secondary interactions, the microstructure of
the polymers, their chemical structure, and the thermody-
namic conditions (temperature, solvent composition, pH,
salt nature) are all important for controlling the conforma-
tion, interchain interaction, and stiffness of the chain, as
well as the solution and gel properties.
[1] G. A. Jeffrey, W. Saenger, Hydrogen Bonding in Biological
Structures, Springer, Heidelberg 1994.
[2] M. Hirrien, J. Desbrieres, M. Rinaudo, Carbohydr. Polym.
1996, 31, 243.
[3] M. Vigouret, M. Rinaudo, J. Desbrieres, J. Chim. Phys. 1996,
93, 858.
[4] J. Desbrieres, M. Hirrien, M. Rinaudo, Relation between
the Conditions of Modication and the Properties of
Cellulose Derivatives: Thermogelation of Methylcellulose,
in: Cellulose Derivatives-Modication, Characterization
and Nanostructures, T. J. Heinze, W. G. Glasser, Eds.,
ACS Symposium Series 688, American Chemical Society,
Washington, DC 1998, p. 332.
Figure 25. Storage modulus (G
0
) and loss modulus (G
00
) for LBG(!G
0
, !G
00
) and the mixture 50/50 LBG/
xanthan(} G
0
, ^G
00
) at 10 g L
1
in water at 25 8C.
608 M. Rinaudo
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