Colloids and Surfaces A: Physicochem. Eng.

Aspects 255 (2005) 181–191

Phase behavior in mixtures of cationic surfactant

and anionic polyelectrolytes
Marko Vincekovića , Marija Bujana , Ivan Šmitb , Nada Filipović-Vincekovićc,∗
a Department of Chemistry, Faculty of Agronomy, 10000 Zagreb, Croatia
bDivision of Materials Chemistry Rud--er Bošković Institute, 10002 Zagreb, Croatia
c Department of Physical Chemistry, Rud--er Bošković Institute, 10002 Zagreb, Croatia

Received 12 October 2004; accepted 19 December 2004

Available online 29 January 2005

Abstract

Interactions in mixtures of n-dodecylammonium chloride (DDACl) and ␬-, ␫- and ␭-carrageenan (CAR) have been investigated by surface
tension, conductometry, light microscopy, light scattering, microelectrophoretic and X-ray measurements. The phase behavior in dilute water
corner includes the regimes of soluble and insoluble complexes. Various polyelectrolyte–surfactant complexes (PSC) were formed at the
air/solution interface and in bulk. The effect of the carrageenan concentration on the critical aggregation concentration (cac) and critical
micelle concentration (cmc) was found to be very weak. The increase of carrageenan charge density increases the cac and cac/cmc ratio, and
shifts the precipitation region towards higher concentration of both components. Factors governing the PSC formation were discussed in terms
of electrostatic interactions, conformation, flexibility of the polyelectrolyte chains and entropy gain due to released carrageenan counterions.
Formation of giant vesicles even at low carrageenan concentration and structural properties of insoluble complexes revealed lamellar ordering
as an important feature of the DDACl/CAR systems.
© 2004 Elsevier B.V. All rights reserved.

Keywords: Dodecylammonium chloride; Carrageenans; Polyelectrolytes; Surfactants; Vesicles

1. Introduction tion [19], hydrophobicity [20], degree of polymerization [21],

From the fundamental and industrial aspects (food pro-
cessing, pharmaceutical formulations, personal care prod-
ucts, paints, etc.), there is a considerable interest in the in-
teractions between the polyelectrolyte and surfactants. Many
reviews and books covering different aspects of their interac-
tions have been published [1–8]. The nature and strength of
the interactions depend on the properties of both the surfac-
tant and the polyelectrolyte, composition of systems, solvent
medium, etc. With respect to the surfactant, structure of hy-
drophobic part and head group structure have been found to be
most important factors [9,10]. Polyelectrolyte properties that
affect these interactions are as follows: the polyelectrolyte
charge density [11–16], chain flexibility [17,18], conforma-
∗ Corresponding author. Tel.: +385 1 4561 111; fax: +385 1 4680 098.
E-mail address: filipovi@irb.hr (N. Filipović-Vinceković).
polyelectrolyte counterion specificity [22], etc.
Polyelectrolyte and oppositely charged surfactant inter-
actions are characterized by the fact that the complexation
takes place in a highly cooperative manner. A strong associa-
tion between a polyelectrolyte and oppositely charged surfac-
tant is primarily of electrostatic origin. Additionally, the hy-
drophobic interaction between surfactant alkyl chains starting
at certain surfactant concentration (termed as critical aggre-
gation concentration, cac) enhances binding of surfactants
to polyelectrolyte. Many experimental and theoretical stud-
ies accepted the structure of the polyelectrolyte–surfactant
complexes (PSC) as micelle-like aggregates residing within
random coil of the polyelectrolyte [8]. The precise structure
of PSC depends on the structure of involved molecules, i.e.
the charge and shape of surfactant molecule as well as on the
characteristic of oppositely charged polyelectrolyte. A vari-
ety of surfactant organizations in conjunction with the poly-

0927-7757/$ – see front matter © 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.colsurfa.2004.12.047
182 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191
electrolyte have been observed (ranging from simple micellar
to cubic, hexagonal, lamellar and even coexisting cubic and
hexagonal) [23–26]. Additionally, the complexation of poly-
electrolytes and oppositely charged colloidal particles has
been recently received great attention due to the importance
of resulting aggregates as a complex vesicle-based structures
[27]. The complexation of polyions with oppositely charged
liposomes leads to the formation of aggregates with two dif-
ferent structural arrangements, whose evolution depends on
the charge and mass ratio [28]. The resulting complexes were
shown to be a new class of colloids. There are many poten-
tial applications of these investigations, in both chemical and
biological areas.
In an earlier paper, we have reported on the phase behavior
of mixtures containing anionic polyelectrolyte, ␫-carrageenan
(IC) and cationic surfactant, n-dodecylammonium chloride
(DDACl) [29]. The results revealed lamellar ordering of the
PSC even in extremely dilute IC solutions. In our present
work, we have extended our previous investigation on the in-
teractions between DDACl and the three differently charged
carrageenans. Due to the presence of sulfate groups, car-
rageenans are strong polyelectrolytes. Their properties in
aqueous solutions are strongly affected by primary differ-
ences in number and position of the sulfate groups within the
repeating galactose units (charge density) as well as by the
amount of anhydro-galactose. Ideally, ␬- and ␫-carrageenan
possess one single or two sulfate groups per disaccharide unit,
whereas in ideal structure, ␭-carrageenan (LC) contains three
sulfate groups. The idealized repeating structures of three car-
rageenans, KC, IC and LC are illustrated in Scheme 1.
Here, we compare three main carrageenan species from
the point of view of their charge under the concentration
conditions from infinite dilution to the phase separation.
Several different methods have been applied for evalua-
tion of interactions and structural characteristics of PSC.
The growth of carrageenan charge density increases the cac
values and shifts the clear/turbid boundary towards higher
polyelectrolyte concentration. The cac increase with increas-
ing charge density of carrageenans instead of its decrease
indicates that chains flexibility and their conformation as
well as counterions entropy gain are important factors in
control of the PSC formation. Although the findings ob-
tained in the dilute regime cannot be generalized to precip-
itation region, one could easily find the correlation between
structural features of soluble PSC and precipitated solid
phase.
Scheme 1. Ideal repeating disaccharide unit of ␬-, ␫- and ␭-carrageenan.
2. Experimental
2.1. Materials and sample preparation
The preparation and purification of DDACl were described
earlier [30]. The carrageenans were purchased from Fluka
and purified by dialysis and subsequent percolation of re-
spective aqueous solutions through an ion exchanger, and
transformed to sodium salts as described previously [19].
Stock solutions were prepared by weight using redistilled
and filtered water (through Millipore Type HA 0.10 ␮m, to
exclude dust before use). The systems were prepared with
a constant carrageenan concentration and variable surfactant
concentration. They were stirred for 5 min and left aside for
equilibration at 30 ◦ C. All mixtures were prepared at least
24 h before measurements.
The stoichiometric compounds (n-dodecylammonium ␬-,
␫- or ␭-carrageenate, DDAKC, (DDA)2 IC, (DDA)3 LC, re-
spectively) were prepared by mixing concentrated aqueous
solutions of n-dodecylammonium chloride and correspond-
ing carrageenan. The coprecipitated NaCl was washed with
cold water. Precipitates were dried in vacuo at room temper-
ature.
2.2. Methods and interpretation of data
Detection and microstructural identification of different
phases were performed by light microscopy (Model DMLS
equipped with a video camera, Leica), X-ray diffraction
(Philips diffractometer with monochromatized Cu K␣ radia-
tion), tensiometry (K8 Interfacial Tensiometer, Krűss), con-
ductometry (MPC 227 Conductivity Meter, Mettler Toledo),
microelectrophoretic and light scattering measurements (Ze-
tasizer Nano ZS, Malvern Instruments). The clear/turbid so-
lution boundary was determined by visual inspection.
The surface properties of DDACl/CAR mixtures have
been investigated by measuring the surface tension, which
shows considerable complex time dependence. In order to
obtain realiable data we used the criterion that the surface
was at equilibrium when any changes in tension were less
than 0.2 mN/m/min.
The cac values were determined by a usual procedure
from the conductivity (κ) versus concentration (c) plots [29].
The cmc values were determined by specific conductivity
measurements as the surfactant concentration at the intersec-
tion of the two linear sections κ versus c curves by a linear
 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191
fitting procedure with correlation factor always better than
0.999.
The average basic lamellar thickness (d/Å) was calculated
from the peaks on the diffractograms by Bragg equation [31]:
d = 1.5418/2 sin θ
where θ is the diffraction angle.
3. Results and discussion
3.1. Phase behavior
Fig. 1a–c shows the dilute corner of the phase diagrams for
DDACl and KC, IC and LC mixtures (DDACl/CAR) ranging
from the dilute-to-turbid and moderately viscous solutions.
Several clear and two turbid regions can be identified as a
function of concentration and molar ratio of both compo-
nents. Full lines marked as 1 denote the cac indicative of
the micelle-like aggregates formation on the polyelectrolyte
chains, whereas lines marked as 2 denote the cmc.
Following the multistep mechanism of interactions [29],
at a surfactant concentration lower than cac (left-hand side
of the line 1), the adsorption of surfactant monomers to car-
rageenan chains are governed by electrostatic interactions in-
volving an ion exchange of carrageenan counterion (sodium)
with DDA+ and formation of negatively charged soluble com-
plexes according to relation:
aDDA+ + n(CAR)−z → [(DDA)a (CAR)n ]−(nz−a) ,
where a is the number of dodecylammonium cations (DDA+ ),
n the number of repeating carrageenan units, CAR−z a car-
rageenan sulfate unit (where z is ideally one, two and three
for KC, IC and LC, respectively).
Beyond the cac further binding of surfactants, governed
by hydrophobic interactions between surfactant alkyl tails,
brought excessive positive charges to bear on the soluble
complexes according to relation:
+ −z
bDDA + n(CAR) → [(DDA)b (CAR)n ] (b−nz)
, (3)
where b denotes aggregation number of micelles residing at
the polyelectrolyte chains.
Fig. 2. Polarization microphotographs of vesicles from mixtures DDACl/CAR with varying c(DDACl) (as indicated) and constant c(CAR)/% = 1 × 10−3 ; bar
is indicated; temperature is 30 ◦ C.
183
(1)
Fig. 1. Partial phase diagram of the DDACl/CAR mixtures was obtained af-
ter 24 h at 30 ◦ C. The heavy full lines correspond to the clear/turbid boundary,
whereas the dashed ones correspond to the coexistence regions of precip-
itated solid phases and vesicles. Regions between heavy and dashed lines
contain large number of giant vesicles. The full lines denoted as 1 and 2
correspond to the changes of the cac and cmc caused by increased CAR
concentration: (a) KC; (b) IC; (c) LC.
Typical polarizing microphotographs of samples with con-
stant carrageenan concentration and increasing DDACl con-
centration (below the cac, between the cac and the cmc, and
beyond the cmc) revealed the formation of giant vesicles with
dimensions of several micrometers (Fig. 2) in all three cases.
Giant vesicles were detected by characteristic maltese crosses
visible under crossed polarizers. The number and size of giant
vesicles strongly depend on the surfactant concentration. The
highest concentration of vesicles was observed in the vicinity
of the precipitation region. Microphotographs from micellar
region reveal the existence of closed thin-walled tubules.
(2) Fig. 3 shows oligovesicular vesicles, giant vesicle and floc-
cules, which coexist inside precipitation region in all three
cases. Microscopic observations between crossed polariz-
ers show that precipitated phases are strongly birefringent.
The appearance of maltese crosses indicates that the vesicles
are densely packed forming flocks of aggregated vesicles. It
seems that the transition from soluble PSC structures to the
precipitate partially proceeds within oligovesicular phase.
3.2. Surface properties
It is well known, that the addition of a polyelectrolyte to
a surfactant of opposite charge causes a synergistic lower-
ing of surface tension at very low surfactant concentration.
184 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191

Fig. 3. Polarization microphotographs of oligovesicular vesicles and floccules from mixtures with c(CAR)/% = 5 × 10−3 and c(DDACl)/mol dm−3 = 1 × 10−3 ;
bar is indicated; temperature is 30 ◦ C.

This lowering arises from a surfactant and poylelectrolyte centration there is no clear evidence of the cac in all surface
enhanced adsorption at the air/solution interface caused by tension isotherms), whereas curves with two plateaus typical
electrostatic interactions between a polyelectrolyte and oppo- for mixtures of a surfactant and oppositely charged polyelec-
sitely charged surfactant headgroup, i.e. by the formation of trolyte appeared at c(CAR) = 1 × 10−4 %. The cross section
polyelectrolyte–surfactant complexes at the air/solution in- thorough turbid zone displays isotherms with initial drop of
terface [32]. New information accessible through X-ray and surface tension at the lowest DDACl concentration and slight
neutron scattering and reflectivity measurements revealed increase up to the value of pure DDACl solutions.
that polyelectrolyte–surfactant complexes are formed at the Typical γ versus log c(DDACl) isotherms of mixtures at
air/solution interface at lower surfactant concentration than two constant KC concentration from clear region are illus-
that seen in bulk phase [33–38]. Further, it is shown that trated in Fig. 4. These two types of curves reflect the phys-
there is a surfactant monolayer at the air/solution interface ical state of the surfactant and carrageenan in bulk. At very
with a thin layer of a polyelectrolyte in the subphase, which low carrageenan concentration the surfactant molecules are
changes to a thicker layer at higher surfactant concentrations. present as monomers, although some association with car-
The precise structure of the sublayers is difficult to determine rageenan chains can occur due to ion exchange of sodium
but it is thought to be composed of surfactant bilayers or mi- ions by surfactant cations below the cac. Beyond the cac the
celles bound to polyelectrolyte that links the sublayers to the formation of micelle-like aggregates on carrageenan chains
surfactant monolayer. leads to a continuous desorption of species present at the inter-
Interactions in the bulk and at the air/solution interface face because of insufficient carrageenan concentration. The
are generally related [39], and one may expect the competi- situation at the air/solution interface is reverse of that below
tion between formation of complexes at surfaces and in the
bulk. In order to correlate surface properties with the bulk
phase it is important to have information of the bulk phase
behaviour. According to the phase diagram (Fig. 1a–c) char-
acteristic carrageenan concentrations have been chosen, two
from the clear region and the third thorough turbid region, i.e.
c(CAR)/% = 1 × 10−5 , 1 × 10−4 and 1 × 10−3 . The solutions
of pure carrageenans do not change the surface tension, thus
suggesting that the carrageenans are not surface active. This
can be ascribed to the hydrophilic character of carrageenans,
i.e. to the presence of sulfate groups. In contrast to the sur-
face tension isotherm of pure DDACl solutions, which dis-
plays the decrease up to the cmc and the constant value in
micellar region, surface tension isotherms of DDACl/CAR
mixtures reveal complex surface properties. Surface tension
measurements of solutions with carrageenans from clear re-
gion display two types of curves; at very low carrageenan
concentration (c(CAR) = 1 × 10−5 %) curves with a shallow
Fig. 4. Variation of surface tension (γ) vs. c(DDACl) at constant
minimum below the cmc were obtained (at this very low con- c(KC) = () 1 × 10−5 %, ( ) 1 × 10−4 %, (䊉) 0%; temperature is 30 ◦ C.
 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191
the cac. The equilibrium between surfactant monomers at the
air/solution interface and monomers in solution is shifted to-
ward the bulk solution [40]. This explains the slight increase
in surface tension close to the values of pure surfactant so-
lution. In micellar region, the surface tension values become
constant but slightly higher than those of the pure DDACl
solution. The value of surface tension of the DDACl solu-
tion is attained only at highest DDACl concentrations (not
shown), indicating that the surface complexes become solu-
bilized in the bulk solution leaving only surfactant monomer
at the air/solution interface.
At higher carrageenan concentration, a break in the sur-
face tension at the critical aggregation concentration corre-
sponds to the formation of significant number of PSC. This
concentration is related to the pronounced hydrophobic inter-
actions between surfactant alkyl chains and structural change
of PSC in bulk. As more DDACl is added, the carrageenan
chains become saturated with surfactant micelles and then
the monomer concentration starts to rise again causing the
surface tension decrease up to the cmc; approaching that of
the pure surfactant solutions.
At the highest carrageenan concentrations (cross section
at c(CAR) = 0.001%), the surface tension isotherms exhibit
the highest surface tension drop, and after that slight rise up
to value of pure DDACl solution (Fig. 5). The increase in
γ instead of its decrease affected by the increase of surfac-
tant concentration can be explained by the formation of giant
vesicles in bulk phase; the solutions become turbid, but no
visible precipitation occurred. In the vicinity of the cmc, the
surfactant monomer concentration starts to increase which
lowers surface tension up to the cmc. Beyond the cmc val-
ues, the solutions become transparent and the surface tension
becomes constant. It is visible from Fig. 5 that when the de-
gree of carrageenan charge is increased the surface tension
decreases.
185
In conclusion, surface complexation strongly depends on
the complexation in bulk. The polyelectrolyte configurations
in bulk and at the air/solution interface are very different and
with no doubt, the structure of complexes formed in the bulk
phase differ from those formed at the surface.
3.3. Bulk properties
3.3.1. Specific conductivity measurements
Specific conductivity measurements reveal that in all in-
vestigated mixtures, the specific conductivity values mea-
sured are smaller than those in the absence of carrageenans,
thus indicating that the interaction between surfactant and
polyelectrolyte starts below 1 × 10−5 mol dm−3 of DDACl
and 1 × 10−7 % of CAR. In pure DDACl solutions, the ␬-c
curve shows two linear regions with an intercept correspond-
ing to the cmc. All specific conductivity plots of DDACl/CAR
mixtures show double breaks, one in the vicinity of the cmc
(0.015 mol dm−3 of DDACl) and the other well below the
cmc. The break at lower concentration can be attributed to
the cac. A typical example of specific conductivity changes
with DDACl concentration for DDACl/KC systems with one
break corresponding to the cac, and the second corresponding
to the cmc is illustrated in Fig. 6a and b.
The changes of the cac and cmc values with carrageenan
concentration are showed in Fig. 1a–c. The cac values dis-
play a negligible increase with the increase of KC or IC con-
centration, whereas this increase is somewhat higher for the
DDACl/LC mixtures. This is in good accordance with the
literature data, which have established that the cac depends
on the polyelectrolyte concentration only weakly [41]. The
addition of carrageenan has not significantly affected the cmc
values of surfactants, i.e. the cmc values are almost the same
as in the single surfactant solutions.
Table 1 shows the increase of cac with a number of sulfate
groups per disaccharide units. At concentrations below the
cac, the interaction between polyelectrolyte and oppositely
charged surfactants is primarily of electrostatic nature and it
is expected that these interactions depend on the charge den-
sity of the polyelectrolyte. The increase in the cac, instead of
the decrease caused by an increase in charge density, i.e. the
opposite trend found in our experiments, indicates that some
other effects have to be considered. The binding of surfac-
tant to a polyelectrolyte is governed by several contributing
factors. Apart from the charge density [12–16,42,43], two
important factors affecting the cac are conformation and flex-
ibility of the polyelectrolyte chains [15,19,44]. It is known
Table 1
Critical aggregation concentration (cac/mol dm−3 ), the cac/cmc ratio, and re-
duction in free energy of micelles when bound to carrageenan (G/kJ mol−1 )
for DDACl/CAR mixtures; temperature is 30 ◦ C
CAR cac × 104 cac/cmc × 102 G
KC 1.48 0.99 −11.6
Fig. 5. Variation of surface tension (γ) vs. c(DDACl), at constant
IC 2.34 1.56 −10.5
c(CAR)/% = 0.001. () DDACl/KC; (
) DDACl/IC; ( ) DDACl/LC; tem-
LC 3.60 2.40 −9.4
perature is 30 ◦ C.
186 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191
Fig. 6. Specific conductivity (κ) vs. c(DDACl) at constant c(CAR)/% = 1 × 10−5 ; at (a) lower and (b) higher c(DDACl); temperature is 30 ◦ C.
that the conformation change in solutions of carrageenans
is a complex process involving coil-to-helix transitions de-
pending on the temperature, carrageenan concentration, ionic
strength and the nature of co- and counterions. Generally, at
low concentration and in the absence of salt, the association
is negligible and carrageenans exist as independent moieties
randomly distributed throughout the system. In dilute concen-
tration region and in the absence of salt, the charge groups
within the polyelectrolyte repeal each other and the poly-
electrolyte chains are in an extended form and quite stiff.
At 30 ◦ C, KC exists in a statistical coil state [45], whereas
almost 90% of IC exists in helical conformation [46]. Molec-
ular characteristics and gelling properties of the carrageenan
family investigated by light scattering and small-angle X-ray
scattering using conventional analysis and molecular model
[47] have clearly shown that the difference in the position of
sulfate groups (rather than the number of sulfate groups per
repeating units) and the presence of the anhydrous residue
are essential for the carrageenan family to assume a helical
conformation. The electric charge of sulfate groups enhances
the extended rod-like structure so that LC assumes an ex-
tended conformation in water. LC is not capable of forming
an ordered domain composed of aligned helices, and as a
consequence gelation takes place at no circumstance.
It has been shown that both the charge density and the dis-
tance between the charges in the carrageenan play a signifi-
cant role in determining the adsorption of surfactant [12,19].
The transformation from coil-to-helix does not change the
charge density, but the length per unit charge is different for
both the coil and the helix form [19]. The distance between the
ionic sites on the carrageenans plays an essential role for the
hydrophobic interactions between the adsorbed molecules,
i.e. the shorter charge–charge distance leads to a higher de-
gree of cooperativity of the adsorption of surfactant to the
carrageenan. Consequently, one may expect higher degree
of interaction between parts of adjacently bound surfactant
molecules. From the other side, the polymer chains in helical
state are less flexible than those in coil state, which leads to
a longer distance between the charged groups on the poly-
electrolyte chains and hence a lower degree of cooperativity
[13,15]. Since the values of the cac reflects the ease of form-
ing the first surfactant aggregate with the polyelectrolyte, the
higher cac in the IC case may be related to a less flexible
IC helix in comparison with the KC existing in flexible coil
conformation. It seems that, due to the higher stiffness of the
IC chains, the distances between the charge groups are of
such a magnitude and their positions are such that they allow
hydrophobic interactions to start at surfactant concentrations
higher than those for KC.
Simple models ignoring the counterions condensation ef-
fects could not explain the increase of the cac with an in-
creased charge density of a polyelectrolyte. Strongly charged
polyelectrolytes have condensed counterions associated with
them that can be released when the surfactant binds to the
polyelectrolyte [48]. The counterions in polyelectrolyte so-
lutions are bound to the polyelectrolyte chains, thereby losing
their translational entropy [49]. When cationic surfactant is
added, the head groups of surfactant are individually bound
to the sulfate groups along the carrageenan chains due to elec-
trostatic attractions, i.e. condensed polyelectrolyte counteri-
ons are replaced by surfactant molecule. Consequently, the
concentration of carrageenan counterion increases in bulk
phase causing the cac to increase as salt is added. At the
same time, no change of surfactant counterion concentra-
tion has been observed indicating that surfactant counterions
are not involved in PSC [50]. Accordingly, in DDACl/CAR
mixtures the entropy gain of the counterions released into
aqueous phase upon PSC formation will increase with car-
rageenan charge. Both, higher releasing of counterions from
highly charged carrageenans and decrease of flexibility out-
weigh the effect of an increase of the charge density on the
cac and delays the occurrence of hydrophobic aggregation
of bound surfactant monomers, i.e. the cac value increases.
Similarly, higher release of sodium ion from the carrageenan
 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191
chains may be used for the explanation of observed weak
change of the cac with increasing polyelectrolyte concentra-
tion.
Wallin and Linse have found that the cac/cmc ratio de-
creases with (i) increasing surfactant tail length [16], (ii) in-
creasing linear charge density of the polyelectrolyte [42], and
(iii) increasing flexibility of the polyelectrolyte [44]. It was
found that the cac/cmc ratio is related to the free energy of
complexation between the micelle and polyelectrolyte in so-
lution. The reduction in the Gibbs free energy of micelles
187
It would be expected that, due to partial neutralization of
negative charges, the electrostatic repulsion would become
weaker and consequently, the electrophoretic mobilities be-
come lower. However, at the lowest DDACl concentration,
somewhat higher negative mobility than that of correspond-
ing carrageenan may be explained by considering the change
in the shape and/or size of complexes, which are additional
factors determining the electrophoretic mobility [52]. A fur-
ther increase of c(DDACl) leads to a higher DDACl binding
and decrease of electrophoretic mobility. The overall par-
when bound to a polymer (G) can be expressed as [51]: ticle charge is reduced due to the electrostatic binding of
ammonium head groups of surfactants to the anionic sul-
G = RT ln(cac/cmc) (4) fate sites along the polyelectrolyte chain. When surfactant
Our results reveal that the cac/cmc ratio increases as the concentration reaches the cac value, PSC crosses zero mobil-
charge density of carrageenan increases (Table 1) and con- ity and becomes positively charged. At the cac, the negative
sequently, the absolute value of G decreases from KC to
LC. The change in the free energy of polyelectrolyte-bound
micelles exhibits a decrease that indicates the growth of the
number of sulfate groups per disaccharide unit decreases the
stability of polyelectrolyte–surfactant complexes.
3.3.2. Microelectrophoretic mobility measurements
The microelectrophoretic mobilities of PSC are shown as
a function of the surfactant concentration in Fig. 7. In the ab-
sence of surfactant and at 1 × 10−5 % of carrageenan solution,
the approximate values of the microelectrophoretic mobility
for KC, IC and LC are −0.23 × 10−4 , −0.75 × 10−4 and
−0.97 × 10−4 cm2 s−1 V−1 , respectively. The carrageenan
chains would display an extended form and at low concen-
trations would usually adopt a random configuration suitably
modified by the repulsive interactions between negatively
charged sulfate groups. Upon addition of the lowest concen-
tration of DDACl, the negative mobility of PSC is some-
what higher than that of corresponding carrageenan chains.
Fig. 7. Variation of the microelectrophoretic mobility (µ± ) of PSC with
c(DDACl) at constant c(CAR)/% = 10−5 ; () KC, (
) IC, and ( ) LC;
temperature is 30 ◦ C.
potential of carrageenan seems to be neutralized by bound
surfactant monomers. This concentration coincides with the
cac determined by conductivity measurements. Further bind-
ing of surfactant monomers brings excessive positive charges
on the PSC. Below the cmc of surfactant, each complex
reaches lower values than the one of pure DDACl micelles
(4.37 × 10−4 cm2 s−1 V−1 ).
Two populations of positively charged particles appear
in micellar region (not shown); the first one showing the
decrease of mobility with surfactant concentration from
7.9 × 10−4 to 6.1 × 10−4 cm2 s−1 V−1 , and the second one
showing the population with almost constant electrophoretic
mobility. Smaller mobility values are close to the one of the
DDACl micelles. It is in accordance with the results obtained
in dextran sulfate-dodecylpyridinium bromide mixtures, in
which two kinds of migrating species have been observed
[53].
3.3.3. Light scattering measurements
Dynamic light scattering measurements reveal a broad size
distribution that corresponds to a widely polydisperse mix-
tures for all carrageenans. The variation of the mean particle
size (d) with the increasing surfactant concentrations is il-
lustrated in Fig. 8. In the absence of surfactant and at used
carrageenan concentrations, the mean particle sizes of KC, IC
and LC are 32, 57 and 72 nm, respectively. The increase of the
charge density increases the electrostatic repulsion between
charged segments and, consequently, the dimension of the
polyelectrolyte chains leading to more open conformation,
i.e. the mean particle size of carrageenan chains increases
with the charge density. When low concentration of DDACl
is added, the mean particle sizes of all carrageenan chains
are somewhat lower than in pure carrageenan solutions. The
decrease of the PSC size with an increase in the DDACl
concentration can be explained by reduction of the interseg-
mental repulsion, i.e. with the progress of surfactant binding,
the electrostatic repulsion between negatively charged sulfate
groups becomes weaker and the polyelectrolyte chains can
be expected to shrink. Diamant and Andelman [41] have ex-
amined the influence of surfactant on the effective interaction
between polyelectrolyte segments and found that the interac-
188 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191
tions would lead to a partial collapse of the polyelectrolyte
chains at the cac. Above the cac, all complexes exhibit sharp
increase in size up to 0.001 mol dm−3 of DDACl; afterwards
this increase is somewhat slower. The increase of PSC size
beyond the cac arises from at least two reasons, i.e. from the
increased molar mass and increased positive charge of PSC. It
seems that the increasing electrostatic repulsion between the
introduced positive charges causes elongation of complexes.
Comparing the size of PSC for different carrageenans reveals
that, although the charge density of KC is lower than IC and
LC, the mean particle size of PSC is the highest. At least two
types of complexes below the cmc have been identified, how-
ever it was not possible to determine the precise structure of
the soluble PSC by techniques used.
Light scattering experiments revealed the appearance of
at least two kinds of particles in micellar region (not shown).
Two or three size distributions are obtained in almost all
mixtures. The fraction containing smallest particles shows
narrow size distribution, whereas fraction containing larger
particles shows relatively wide distribution, which as DDACl
concentration increases displays two peaks. A peak analysis
per number indicates that almost 99% of particles correspond
to the smallest particles. The average size of the smallest par-
ticles is close to the actual diameter of the DDACl micelles
(≈3.6 nm) indicating the formation of free DDACl micelles.
Larger particles ranging from 500 nm to several microns in
their average size can be attributed to the PSC. Small number
of the PSC present, the decrease of microelectrophoretic mo-
bility and slight increase in average size suggest that at this
very low carrageenan concentration, the intrapolyelectrolyte
complexes are formed in all three cases.
In conclusion, at least two types of complexes below
the cmc have been identified. It is obvious, that the struc-
tures of PSC differ below and above the cac. Initially,
carrageenan–surfactant complexes are negatively charged
and their size decreases due to binding of surfactant cation
Fig. 8. Variation of the mean particle size (d) with c(DDACl) at constant
c(CAR)/% = 10−5 ; () KC, (
) IC, and ( ) LC; temperature is 30 ◦ C.
to sulfate groups on the carrageenan backbone, i.e. shielding
of the electrostatic repulsion between charged carrageenan
sites leads to shrinking of the complexes. Beyond the cac,
the complexes are positively charged and their size increases
up to the micellar region due to elongation and/or increased
molar mass. In micellar region, when the carrageenan con-
centration is small, the majority of the surfactant exists as
free micelles in solution and all the carrageenan chains are
saturated with micelle-like clusters.
3.4. Solid state properties
The concentration of polyelectrolyte is an important pa-
rameter in the PSC formation. Thus, at low polyelectrolyte
concentrations, the intrapolymer complexes are prevailing
form [54]. When the concentration of polyelectrolyte is
high enough, the interpolymer complexes might be formed,
i.e. with an increasing in polyelectrolyte concentration, the
intrapolymer complexes transform into interpolymer com-
plexes, which aggregate more readily and thus lead to pre-
cipitation. However, the microscopic mechanism responsible
for these transitions has not been not fully understood yet. The
number and size of PSCs grow with an increase in polyelec-
trolyte concentration, and finally the liquid/liquid separation
proceeds into the solid/liquid separation or precipitation. The
elemental analysis of precipitates revealed formation of an al-
most stoichiometric solid phase (with respect to the charge
per disaccharide units) according to relation:
zDDA+ + (CAR)−z → (DDA)z CAR, (5)
where z denotes 1, 2 or 3 for KC, IC and LC, respectively.
(DDA)z CAR is n-dodecylammonium carrageenate. The for-
mation of stoichiometric solid phase is in accordance with
the literature data according to which the ionic surfactant
molecules bind in stoichiometric proportions to oppositely
charged sites on polyelectrolyte chains [55].
Most of the polyelectrolyte–surfactant complexes studied
so far showed lamellar ordering [56–58], hexagonal arrange-
ment [59] or a cubic system [60]. To investigate structural
characteristics of dried (DDA)z CAR, we have studied sam-
ples by a wide angle X-ray diffraction. Fig. 9 shows X-ray
diffractograms of (DDA)z CAR, which display two crystalline
reflections in the small-angle region and one expressive dif-
fuse diffraction maxima (typical for amorphous phase) in a
wider-angle region. Such X-ray patterns and the 1:2 relation-
ship of the reciprocal interplanar spacing of two small-angle
reflections are typical for lamellar (smectic) mesophase with
disordered dodecylammonium chains (with cationic surfac-
tant head groups electrostatically bound to sulfate groups)
within the ordered carrageenan chains filling the space be-
tween. The two diffraction lines are indexed as 0 0 l (ac-
tually as 0 0 1 and 0 0 2) and their interplanar spacing for
DDAKC and (DDA)2 IC are d0 0 1 = 36.0 Å, d0 0 2 = 18.4 Å,
while (DDA)3 LC shows d0 0 1 = 34.6 Å and d0 0 2 = 18.0 Å.
The obtained values of the long spacing (d0 0 1 ) are in good
 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191
Fig. 9. X-ray diffractograms of solid n-dodecylammonium carrageenate; (a)
DDAKC, (b) (DDA)3 LC, and (c) (DDA)2 IC.
agreement with the results performed by the small-angle X-
ray scattering (SAXS) and small-angle neutron scattering
(SANS) on carrageenan gels [56–58].
Weak and broadened second diffraction line in n-
dodecylammonium carrageenates (of relative intensity
I/I1 ∼ 5) may indicate rather irregular boundaries between
bilayers or partly bilayered lamellae. Fully extended lengths
of the dodecyl chain (nC = 12) are of about 16.7 Å [61]. Con-
sidering the values of ionic radii of ammonium and sulfate
groups (2.5 and 4 Å [62]) as well as the fact that disordered
hydrocarbon chains are possibly neither fully extended nor
upright, the values of interplanar spacing d0 0 1 could corre-
spond to bilayer thickness of basic lamellae.
In all solid complexes, relatively strong and narrow first
peak diffraction maximum indicates multilayered stacking of
lamellae in mesophase domains. Stronger reflection for the
(DDA)3 LC (Curve b, Fig. 9) than those of the DDAKC and
(DDA)2 IC (Curves a and c, Fig. 9), indicates better lamel-
lar stacking of the (DDA)3 LC. Higher charge density of
(DDA)3 LC may contribute to higher intensity of this peak,
too. The degree of disorder (∆/ā) was calculated from the
half-maximum width β of first reflection according to equa-
tion proposed by Vainshtain [63]. Relatively low ∆/ā val-
ues for DDACl/CAR samples (∆/ā ∼ 0.115) in comparison
to literature data for carrageenan/cetylpyridinium chloride
(∆/ā = 0.14–0.16) [57] indicate somewhat enhanced lamellar
stacking.
The intensive diffraction halo around 2θ/◦ = 20, i.e. at
≈4.4 Å, which is characteristic for disordered liquid-like
conformation of liquid paraffins [64] suggests a short-range
189
order of hydrocarbon chains in bilayers only. The same
shape and half-maximum width for DDAKC and (DDA)2 IC
(β = 6.4◦ 2θ) and somewhat narrower half-maximum width
for (DDA)3 LC (β = 6.0◦ 2θ) indicate slightly enhanced local
ordering of alkylammonium chains within the (DDACl)3 LC.
(DDACl)3 LC has the highest charge density and the high-
est amount of bound surfactant, which enhance the molecu-
lar packing in comparison to two less charged carrageenans.
The increased ordering results from higher electrostatic and
hydrophobic interactions. Additionally, the diffractogram of
(DDA)3 LC exhibits slight diffuse amorphous halo at low
Bragg’s angles with the maximum at ∼9.5 Å. For the mo-
ment we have no explanation for the appearance of this halo.
The results of the X-ray analysis reveal the lamellar pe-
riodicity in mesostructural domains (mesomorphites in anal-
ogy to crystallites) consisting of surfactant bilayers between
polyelectrolyte backbones. It seems that in the case of stiff
carrageenan chains the most favourable orientation of surfac-
tant molecule in the solid phase would be in the form of bilay-
ers between carrageenan chains Surfactant bilayers between
polyeletrolyte chains in a lamellae are disordered domains
with slightly different local ordering. Stronger electrostatic
interactions with oppositely charged surfactant head groups
and oppositely charged carrageenan sulfate groups enhance
both, local ordering of bilayers and regular stacking of lamel-
lae. This finding is in accordance with the results obtained
in literature [56–58]. The effect of ordering is stronger the
more the carrageenan chains are charged, i.e. the presence of
charges promote layering of the polyelectrolyte backbones.
4. Conclusions
The investigations of aqueous mixtures of the cationic sur-
factant, n-dodecylammonium chloride and anionic polyelec-
trolyte, carragenans difering in charge density, reveal that for-
mation of various complexes at the air/solution interface and
in bulk is controlled by several contributing factors such as
electrostatic interactions, hydrophobicity of surfactants, con-
formation and flexibility of chains as well as entropy change
due to the release of polyelectrolyte counterions. The final
size and structures of complexes depend upon the relative
magnitudes of these factors.
The configuration and composition of PSC at the
air/solution interface and in bulk are different, i.e. the struc-
ture of complexes formed in bulk phase differs from those
formed at the air/solution interface. While different highly
surface active complexes form at the air/solution interface,
there are at least two kinds of bulk complexes, the first be-
low the cac and the second between the cac and the cmc.
The variation of complex size and charge with surfactant
concentrations can be attributed to the rearrangement of
structure, which finally results in the formation of giant and
oligovesicular vesicles indicating bilayer ordering. With in-
creasing carrageenan concentration, the soluble intrapolymer
complexes change to interpolymer complexes, which subse-
190 M. Vinceković et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 255 (2005) 181–191
quently aggregate in order to form the microphase separated
domains.
Although we cannot generalize the findings in the dilute
regime to higher concentration and vice versa, the appearance
of giant and oligovesicular vesicles and structural character-
istics of insoluble PSC indicate lamellar ordering of at least
positively charged soluble PSC above the cac. Since, by bind-
ing surfactant ions to the polyelectrolyte chains, the complex
has been formed, the structure of surfactant aggregates will
significantly depend on the flexibility of the polyelectrolyte
chain. The carrageenans as rather stiff polyelectrolyte chains
induce a lamellar-like arrangement of surfactant, which en-
ables them to remain in a more or less extended conforma-
tion. Most favorable orientation of surfactant molecules in
the complexes would be in the form of bilayers, thus allow-
ing the formation of vesicles. It is in accordance with the
results of Kabanov et al. [65], who have shown that the com-
plexes formed by diblock copolymers and single-tail surfac-
tant in aqueous solutions arrange spontaneously into vesi-
cles. They have pointed out that this unique self-assembly
behaviour makes these systems promising for use in pharma-
ceutics. Talmon and co-workers have shown the coexistence
of several microstructures (bilayer fragments, small vesicles
and disc-like aggregates) in cationic polymer and an anionic
surfactant as the result of a complex interplay between hy-
drophobic, electrostatic and ionic strength effect [66]. They
have concluded that more work is needed to understand better
the very complex self-association process in ionic surfactant
and oppositely charged polyelectrolyte systems.
The appearance of giant vesicles even at low carrageenan
concentrations and structural properties of insoluble com-
plexes reveal lamellar ordering as an important feature of
the DDACl/CAR systems. The increase of the carrageenan
charge density improves the ordering in insoluble PSC as a
result of increased electrostatic and hydrophobic interactions
between carrageenan chains and DDACl.
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