Polymer International Polym Int 54:659–666 (2005)

DOI: 10.1002/pi.1738

Preparation and characterization

of chitosan-based dispersions
Cypriano G da Trindade Neto,1 Ana LP Fernandes,1 Ana IB Santos,1
Waldenice A Morais,1 Marcos VM Navarro,2 Tereza NC Dantas,1 Márcia R Pereira1
and José LC Fonseca1∗
1 Departamentode Quı́mica, Universidade Federal do Rio Grande do Norte Lagoa Nova, Natal, CP 1662, RN 59078-970, Brazil
2 Departamentode Farmácia, Universidade Federal do Rio Grande do Norte Av Gal Cordeiro de Farias, S/N, Petrópolis, Natal, RN
59010-180, Brazil

Abstract: Complexation of chitosan in aqueous solutions by low molecular weight electrolytes is one of
the simplest methods for the preparation of aqueous chitosan dispersions. In this work, the influence of
storage time, sulfate concentration, method of preparation and surfactant content on some properties of
the resultant chitosan dispersions (turbidity, viscosity and zeta potential) was analyzed. Turbidimetry was
adequate to monitor the formation of particles, while viscometry was suitable to monitor changes in the
dispersing phase. An analysis of the properties of these systems, mainly in terms of particle–particle and
macromolecule–macromolecule interactions was carried out.
 2004 Society of Chemical Industry

Keywords: chitosan; particles; zeta potential; controlled release

INTRODUCTION of several biologically active substances: Muller and

Chitosan is a biopolymer which is a derivative of chitin,
a mucopolysaccharide, which is abundantly present
in the nature as the supporting material of crus-
taceans and insects.1 This macromolecule has great
potential in biomedical and pharmaceutical applica-
tions, because of its exceptionally low immunogenicity,
despite the presence of nitrogen in its macromolecular
chain.2 Chitin is structurally similar to cellulose, with
an amino polysaccharide having acetamide groups at
the C2 positions in place of hydroxyl groups, chi-
tosan being obtained through N-deacetylation of these
groups.3
Different applications have been found for this
polymer: in the field of membranes, for instance,
the use of this biomaterial has been described in
the process of permeation of salt solutions,4 as
well as in membranes directed to drug permeation.5
The outstanding biocompatibility of this material has
stimulated some efforts in the direction of chitosan use
as a material for implants, artificial skin, and wound
dressing.6 – 10 In the field of controlled drug release, this
macromolecular material has been used in micro- and
nanosized particle systems for the controlled release
Jacobs11 showed that, when incorporated to chitosan,
the bioavailability of buparvaquone, an antibiotic
which is poorly soluble in water, may be strongly
increased, especially at the level of nanosized systems.
Chitosan particles may be obtained using meth-
ods which involve organic solvents or high energy-
consuming steps, such as the spray-drying method.12,13
However, more interesting methods of obtaining chi-
tosan micro- and nanoparticles have been found, The
main advantage of these methods is that the particles
are obtained from aqueous chitosan solutions by reac-
tions or electrostatic interactions which may involve
covalent crosslinking, complexation or precipitation
from aqueous solutions.14 A primary method of par-
ticle preparation is the neutralization of the weakly
acidic NH3 + groups, present in the chain via the
addition of NaOH; the polarity of the neutralized
macromolecule strongly decreases, resulting in its col-
lapse (precipitation) from the highly polar aqueous
environment.15,16 Covalent crosslinking is usually car-
ried out through the reaction between glutaraldehyde
and non-protonated NH2 groups from the chitosan
backbone,17 a synergistic approach being possible

∗ Correspondence to: José LC Fonseca, Departamento de Quı́mica, Universidade Federal do Rio Grande do Norte Lagoa Nova, Natal, CP
1662, RN 59078-970, Brazil
E-mail: jlcfonseca@uol.com.br
Contract/grant sponsor: Conselho Nacional de Desenvolvimento Cientı́fico e Tecnológico (CNPq)
Contract/grant sponsor: Ministério da Ciência e Tecnologia (MCT)
Contract/grant sponsor: Fundação Coordenação de Aperfeiçoamento de Pessoal de Nı́vel Superior (CAPES)
Contract/grant sponsor: Banco do Nordeste
Contract/grant sponsor: PróReitoria de Pesquisa e Pós-Graduação da Universidade Federal do Rio Grande do Norte (PPPgUFRN)
(Received 15 April 2004; revised version received 27 August 2004; accepted 1 September 2004)
Published online 24 December 2004
 2004 Society of Chemical Industry. Polym Int 0959–8103/2004/$30.00 659
CG de Trindade Neto et al
through the combination of this method and precipi-
tation via aqueous NaOH.18 The main problem with
covalent crosslinking is that glutaraldehyde, in some
cases, may immobilize the substance to be released,19
not to mention that it has been reported that resid-
ual glutaraldehyde may cause mucosal irritation.20
Complexation with electrolytes is another way to
obtain chitosan particles, through ionic crosslinking
of chitosan, arising from the electrostatic interaction
between NH+ 3 groups in the chitosan backbone and
polycharged anions.21 Regarding polycharged anions
which are also macromolecules, namely polyelec-
trolytes, poly(acrylic acid) and its derivatives have
been widely used in the preparation of chitosan parti-
cles of different sizes.22 An attractive approach is the
use of polycharged anions from low molecular weight
electrolytes, a procedure that has been used in the case
of tripolyphosphate (TPP), resulting in particles with
different forms.23 – 27 The same method is applicable
in the case of sulfate anions.28 – 31
The objective of this work is to prepare chitosan
dispersions via sulfate ionic crosslinking and to study
the influence of sulfate and surfactant concentrations,
as well as mechanical stirring and sonication, on the
properties of these dispersions using turbidity, zeta
potential, and viscosity measurements.
EXPERIMENTAL
Materials
Acetic acid (PA, Reagen, Brazil), K2 SO4 (PA,
Labsynth, Brazil), and the nonionic surfactant
polyoxyethylene-sorbitan monooleate (Tween 80,
Sigma, USA, Fig 1) were used without further purifi-
cation. This surfactant was chosen in order to
stabilize the chitosan dispersions, due to its well-
known good compatibility with biological systems,
as reported in the literature.32 Chitosan (Polymar
Ltda, Brazil) was used in this work with a deacety-
lation degree of ca 85 %, and a viscosity molecular
weight Mv ca 2.9 × 105 g mol−1 (determined using the
Mark–Howink–Sakurada equation from viscometric
data33 ). All chitosan solutions were filtered using a
Millipore Millex filter, prior to use. In all experiments,
bidistilled water was used.
Preparation of dispersions
Chitosan dispersions were prepared by the addition of
aqueous 100 g l−1 K2 SO4 to chitosan solutions in the
presence of surfactant (Tween 80). Three methods of
dispersion preparation were used:
Figure 1. Structure if nonionic surfactant Tween 80.
660
(1) An appropriate volume of a 100 g l−1 K2 SO4
solution was added at a volume rate of flow
of 1.5 ml min−1 (using a burette) to a 250-ml
Erlenmeyer flask containing 100 ml of a 2.5 g l−1
chitosan solution (the solvent being aqueous acetic
acid at 20 g l−1 ) and a given amount of surfactant.
Throughout the addition, the Erlenmeyer flask
was immersed in an ultrasonic bath (Transsonic
model T460, Germany, tank volume 2.75 l,
frequency of operation 45 Hz, power 170 W), at a
temperature of 30 ◦ C. The resultant dispersion
was withdrawn from the ultrasonic bath after
20 min (this time takes into account the time
involved in the addition of the K2 SO4 solution).
(2) The same procedure described in (1), with the
difference that the entire process was carried out
under mechanical stirring. The stirrer was driven
by a servo motor (9 V AC, the same as used in
turntables) which had a nominal rotation N of
2100 rev min−1 . The impeller had a diameter D
of 2 cm, so that the resulting nominal Reynolds
DN2 ρ 2 · (2100/60)2 · 1
number was Re = µ ≈ 0.01 ≈
−3
2 × 10 , where ρ = 1 g cm and µ = 0.01 P are,
5
respectively, the approximate density and viscosity
of the solutions.
(3) The same procedure described in (2). However
there was only mechanical stirring (here, sonica-
tion was not employed).
The variables studied were:
• The final molar concentration of K2 SO4 (ie after
the addition was carried out). In this case, a fixed
surfactant concentration of 10 g l−1 was used. This
concentration was chosen because it is midway
between the maximum (20 g l−1 ) and the minimum
(0) used in this work
• The final mass/volume concentration of surfac-
tant. Here, K2 SO4 concentration was fixed at
0.0058 mol l−1 . This value was chosen because the
zeta potential was midway between the maximum
and the minimum value found in this work
Viscometry, turbidimetry and zeta potential analysis
were carried out after the dilution of the resultant
dispersions to 10 % of the initial chitosan concen-
tration (ie 0.25 g l−1 ). This dilution was carried out
using a 20 g l−1 acetic acid aqueous solution. At these
low concentrations, all the resultant dispersions had
Newtonian behavior.
Turbidimetry
Turbidimetry was carried out using a Hach tur-
bidimeter (model 2100P, USA). The instrument was
equipped with a tungsten-filament lamp and a 90 ◦
detector to monitor scattered and transmitted light.
The instrument’s microprocessor calculated the ratio
of the signals from the 90 ◦ and transmitted light detec-
tors, correcting interferences from color and/or light
absorbing materials. All the measurements were taken
at a temperature of 25 ± 1 ◦ C.
Polym Int 54:659–666 (2005)

Viscometry
Viscometry of the resultant dispersions was carried
out using a Ubbelohde viscometer model 0B,
previously calibrated with various different fluids in
a thermostatic bath. The temperature used in these
experiments was (30.00 ± 0.05) ◦ C.
Electrophoretic mobility and zeta potential
measurements
The electrophoretic mobility measurements, µE ,
were carried out using a Zeta-Meter System 3.0+
(Zeta-Meter Inc, USA). The zeta potentials of the
dispersions, ζ , were calculated from µE by employing
the Smoluchowski relationship:34
µE η
ζ =
ε0 ε
where ε0 is the permittivity of vacuum, ε is the
dielectric constant of water, and η is the viscosity of
the medium. In these measurements, the temperature
was 25 ± 2 ◦ C.
RESULTS AND DISCUSSION
The addition of K2 SO4 solution without sonication
resulted in dispersions with unreproducible charac-
teristics. The addition of the K2 SO4 solution under
sonication, however, gave reproducible results (with
and without the use of mechanical stirring). Sonication
was, therefore, crucial to the preparation of well-
defined particle systems. The importance of sonication
in processes which involve precipitation/crystallization
of different materials from their aqueous solutions
has already been reported in the literature. It has
been have found that, inorganic powders are obtained
(a)
100
80
τ (NTU)
60
40
20
0 0
0 0.003 0.006 0.009
−1
CK2SO4 (mol l )
Figure 2. Turbidity τ of chitosan dispersions as a function of K2 SO4 concentration, CK2 SO4 . (a) Without mechanical stirring. (b) With mechanical
stirring. Squares: after dispersion preparation. Circles: 24 h after preparation. Triangles: 48 h after preparation. Diamonds: 72 h after preparation.
NTU = nephelometric turbidity unit.
Polym Int 54:659–666 (2005)
Chitosan-based dispersions
by precipitation from pure aqueous solvent, the
occurrence of powders with finer granulometry was
associated with the sonicated-assisted breakage of
physical interactions between particles.35,36
In the next sections the results obtained for
sonicated systems with and without mechanical
stirring are reported and discussed.
Salt concentration effect
The relationship between the turbidity of non-stirred
dispersions and K2 SO4 concentration can be seen in
Fig 2. The increase in K2 SO4 concentration results
in an increase in turbidity, certainly due to the
ionic crosslinking of a higher number of chitosan
macromolecules. In the case of mechanically stirred
dispersions, there is also an increase in turbidity,
(1)
although it is less evident; it must be due to the
dispersive action of mechanical stirring on the forming
particles (in contrast with shear-induced flocculation,
which would make turbidity increase). Turbidity
increased from the first day, indicating that particles
were still being formed after the end of the process
or/and there was particle aggregation with time.
Figure 3 depicts the relationship between viscosity
and K2 SO4 concentration for the particle systems.
As salt concentration increases, the viscosity of the
dispersions decreases in both cases (with and without
mechanical stirring). That could be explained using
the Einstein equation, which describes the viscometric
behavior of colloidal dispersions:37 – 41
η
= 1 + 2.5φ + O(φ 2 ) (2)
ηo
where η is the viscosity of the colloidal dispersion,
η0 is the viscosity of the dispersing phase, and φ
is the volumetric fraction of the dispersed phase.
(b)
100
80
τ (NTU)
60
40
20
0.012 0 0.003 0.006 0.009 0.012
−1
CK2SO4 (mol l )
661
CG de Trindade Neto et al
(a) 1.3
1.2
η (10−3 Pa s−1)
1.1
1 1
0.9
0.8
0 0.003
CK
2
SO (mol l )
4
Figure 3. Viscosity η of chitosan dispersions as a function of salt concentration, CK2 SO4 . (a) Without mechanical stirring. (b) With mechanical
stirring. Squares: after dispersion preparation. Circles: 24 h after preparation. Triangles: 48 h after preparation. Diamonds: 72 h after preparation.
The dispersing phase (which would contain water,
acetic acid and chitosan macromolecules that have
not collapsed yet) can be thought of as a continuous
medium, the chitosan solid particles being dispersed
within it. Particle formation would then influence
dispersion macroscopic viscosity in two ways: (1) the
effect of increasing the volume fraction of particles
would increase the dispersion viscosity and (2) the
effect of decreasing chitosan concentration in the
dispersing phase would decrease the viscosity of this
phase, contributing to the decrease of the overall
macroscopic viscosity. It seems that the second
contribution is more important to the dispersion
viscosity. Both solid chitosan particles and solubilized
macromolecules can also be thought of as the dispersed
phase in a dispersing phase consisting of water, acetic
acid and K2 SO4 . The addition of K2 SO4 would then
result in a decrease in the volume fraction of the
dispersed phase in two ways:
(1) Through the polyelectrolyte effect:42 the addition
of K2 SO4 to the medium would shield the pos-
itive charges in the chitosan chains, decreasing
the repulsion between them and, as a conse-
quence, the dimensions of macromolecules still
solubilized, resulting in the decrease of viscosity;
(2) The formation of particles through ionic crosslink-
ing of positively charged chitosan macro-
molecules. In this case, the decrease in φ would
be more drastic.
It can be seen that the viscosities of stirred sam-
ples were slightly lower than those of the dispersions
made without mechanical stirring. This suggests that
shearing coming from stirring may break physical
interactions between chitosan molecules, disfavoring
662
(b) 1.3
1.2
η (10−3 Pa s−1)
1.1
0.9
0.8
0.006 0.009 0.012 0 0.003 0.006 0.009 0.012
−1
CK SO4 (mol l−1)
2
the presence of macromolecule–macromolecule inter-
actions in solution, and decreasing viscosity, as has
been observed in some biopolymer solutions.43 The
possible causes for the decrease in viscosity with
time in both cases could be that solubilized chitosan
macromolecules underwent hydrolysis44 and/or that
the process of particle formation was still going on
during this time, as suggested by turbidity data.
Figure 4 shows the values of zeta potential, ζ , as
a function of salt concentration for the dispersions
obtained. It can be seen that the increase in salt
concentration resulted in a decrease in values for
the zeta potential. Bearing in mind that chitosan
is a cationic polyelectrolyte, surface charging can
be expected to be positive and, as a consequence,
zeta potential values as well.45 The incorporation of
SO4 2− as the ionic crosslinking agent into the particles
would decrease zeta potential. In terms of ζ , there
was not a great difference between stirred and non-
stirred systems. In the case of stirred dispersions,
the systems with higher concentration of salt could
not be observed using the magnification provided
by the zetameter optical microscopy (although they
were turbid, indicating the presence of a dispersed
solid phase), which supports turbidity-correlated data
analysis which indicated a lower particle size for the
stirred dispersions. The values of zeta potential of the
stirred dispersions do not increase with time as much
as the dispersions obtained without stirring, indicating
that, in the latter case, the occurrence of adsorption of
chitosan macromolecules on the particle surface after
dispersion preparation is more noticeable.
Surfactant concentration effect
Figure 5 shows the influence of surfactant content on
the turbidity of chitosan dispersions with a K2 SO4
Polym Int 54:659–666 (2005)
 Chitosan-based dispersions

(a) (b)
40 40

35 35
ζ (mV)

ζ (mV)
30 30

25 25

20 20

0.003 0.006 0.009 0.012 0.003 0.006 0.009 0.012

CK SO4 (mol l−1) CK SO4 (mol l−1)
2 2

Figure 4. Zeta potential ζ of chitosan dispersions as a function of salt concentration, CK2 SO4 . (a) Without mechanical stirring. (b) With mechanical
stir ring. Squares: after dispersion preparation. Circles: 24 h after preparation. Triangles: 48 h after preparation. Diamonds: 72 h after preparation.

(a) (b)
35 35

30 30

25 25
τ (NTU)

τ (NTU)

20 20

15 15

10 10

5 5

0 5 10 15 20 0 5 10 15 20
CS (g l−1) CS (g l−1)

Figure 5. Turbidity τ of chitosan dispersions as a function of surfactant concentration, CS . (a) Without mechanical stirring. (b) With mechanical
stirring. Squares: after dispersion preparation. Circles: 24 h after preparation. Triangles: 48 h after preparation. Diamonds: 72 h after preparation.
NTU = nephelometric turbidity unit.

concentration of 0.0058 mol l−1 . It can be seen that,
in the case of the mechanically stirred dispersions,
turbidity is much lower, which is the same effect that
occurred in the case of different salt concentrations.
This result corroborates the results obtained with the
variation of K2 SO4 concentration. In this case, the
increase in turbidity with time is much more evident
in the case of the stirred dispersions, indicating that
stirring leaves the dispersions in a sort of metastable
state. As the dispersions stand at room temperature,
their turbidity values tend to the values of the
non-stirred ones.
Figure 6 depicts the influence of surfactant concen-
tration on the viscosity of chitosan dispersions with a
fixed K2 SO4 concentration of 0.0058 mol l−1 . In the
case of the dispersions without stirring, it can be seen
that, initially, there is a decrease in viscosity as the
surfactant content is increased. Since the surfactant
is soluble in water, it can decrease the solubility of
ionically crosslinked chitosan in water, favoring solid

Polym Int 54:659–666 (2005) 663

CG de Trindade Neto et al

(a) (b)

1.04 1.04

1 1
η (10−3 Pa s−1)

η (10−3 Pa s−1)
0.96 0.96

0.92 0.92

0 5 10 15 20 0 5 10 15 20
Cs (g l−1) Cs (g l−1)

Figure 6. Viscosity η of chitosan dispersions as a function of surfactant concentration, CS . (a) Without mechanical stirring. (b) With mechanical
stirring. Squares: after dispersion preparation. Circles: 24 h after preparation. Triangles: 48 h after preparation. Diamonds: 72 h after preparation.

(a) 50 (b) 50

45 45

40 40

35 35
ζ (mV)

ζ (mV)

30 30

25 25

20 20

15 15

10 10
0 5 10 15 20 0 5 10 15 20
CS (g l−1) C S (g l−1)

Figure 7. Zeta potential ζ of chitosan dispersions as a function of surfactant concentration, CS . (a) Without mechanical stirring. (b) With mechanical
stirring. Squares: after dispersion preparation. Circles: 24 h after preparation. Triangles: 48 h after preparation. Diamonds: 72 h after preparation.

particle formation, resulting (as previously discussed)
in the decrease of macroscopic dispersion viscosity.
Conversely, surfactant molecules could also interact
with hydrophobic sites in the chitosan molecules, sta-
bilizing isolated macromolecules in solution, resulting
in an opposite effect (increase in viscosity). The first
effect seems to be more important up to a surfactant
concentration of 15 g l−1 , the second one being pre-
dominant at higher concentrations. Stirring seems to
compensate the destabilizing effect of the surfactant:
it can be seen that, when under stirring, viscosity
continuously increases as surfactant concentration is
increased. Another reason for the decrease in viscos-
ity (in the case of mechanically stirred dispersions)
could be the destruction of chitosan macromolecular
agglomerates by shear, resulting in a dispersing phase
with lower viscosity, as previously pointed out in the
case of the K2 SO4 -related experiments.
Figure 7 shows the effect of surfactant concentration
on the values of zeta potential, ζ , for dispersions

664 Polym Int 54:659–666 (2005)


with and without mechanical stirring. In both cases
ζ values did not change with increase in surfactant
concentration. This is consistent with the fact that
the surfactant used in this work has a nonionic
nature. Surface charge, however, slightly changed
with stirring: although surfactant concentration does
not seem to influence surface charge, stirring slightly
decreases zeta potential values, perhaps because of an
increase in surface ionic crosslinking by sulfate ion, as
a result of a decrease in particle size
CONCLUSION
Dispersions prepared by the addition of potassium
sulfate aqueous solutions to chitosan–acetic acid
aqueous solutions have their properties influenced by
the following variables: method of preparation, salt
concentration, surfactant concentration and storage
time. In the method of preparation, sonication
plays a decisive role in the reproducibility of the
experiments, certainly because of its activity as a
particle–particle interaction breaker, resulting in more
finely divided suspended particles. Mechanical stirring
also decreased particle size, although one could not
obtain reproducible results if mechanical stirring were
not associated with sonication. The occurrence of
particle formation can be directly monitored by
turbidity measurements, while viscosity measurements
are useful to monitor the dispersing phase (which
was found to be the most important factor in terms
of contribution to macroscopic viscosity). Increase
in salt concentration resulted in an increase in
turbidity, resulting from an increase in particle
collapse from sulfate complexation of chitosan. A
decrease in turbidity with mechanical stirring may
be associated with its effect in the destruction of
particle agglomerates by shear; this very shear may
also be associated to the decrease in macroscopic
viscosity due to the destruction of polymer–polymer
interactions in solution (dispersing phase). Turbidity
and zeta potential measurements suggests that particle
formation still goes on when the samples rest, after
preparation.
ACKNOWLEDGEMENTS
The authors thank Brazil’s Conselho Nacional de
Desenvolvimento Cient ı́fico e Tecnológico (CNPq),
Ministério da Ciência e Tecnologia (MCT), Fundação
Coordenação de Aperfeiçoamento de Pessoal de
Nı́vel Superior (CAPES), Banco do Nordeste,
and PróReitoria de Pesquisa e PósGraduação da
Universidade Federal do Rio Grande do Norte
(PROPESQ-UFRN) for financial support during the
course of this work.
REFERENCES
1 Tolaimate A, Desbrières J, Rhazi M, Alagui A, Vincendon M
and Vottero P, On the influence of deacetylation process on
Polym Int 54:659–666 (2005)
Chitosan-based dispersions
the physicochemical characteristics of chitosan from squid
chitin. Polymer 41:2463–2469 (2000).
2 Kumar MNVR, A review of chitin and chitosan applications.
React Funct Polym 46:1–27 (2000).
3 Kurita K, Controlled functionalization of the polysaccharide
chitin. Progr Polym Sci 26:1912–1971 (2001).
4 de Vasconcelos CL, Rocha ANL, Pereira MR and Fonseca JLC,
Electrolyte diffusion in a chitosan membrane. Polym Int
50:309–312 (2001).
5 Rocha ANL, Fonseca JLC and Pereira MR Permeation of drugs
in chitosan membranes. J Appl Polym Sci, 84:44–49 (2002).
6 Khor E and Lim LY, Implantable applications of chitin and
chitosan. Biomaterials 24:2339–2349 (2003).
7 Wang XH, Ma JB, Wang YN and He BL, Bone repair in radii
and tibias of rabbits with phosphorylated chitosan reinforced
calcium phosphate cements. Biomaterials 23:4167–4176
(2002).
8 Suzuki T, Muzushima Y, Umeda T and Ohashi R, Further
biocompatibility testing of silica-chitosan complex membrane
in the production of tissue plasminogen activator by epithelial
and fibroplast cells. J Biosci Bioeng 88:194–199 (1999).
9 Kojima K, Okamoto Y, Miyatake K, Tamai Y, Shigemasa Y
and Minami S, Optimum dose of chitin and chitosan for
organization on non-woven fabric in the subcutaneous tissue.
Carbohydr Polym 46:235–239 (2001).
10 Mi FW, Tan, YC, Liang HF and Sung HW, In vivo biocompat-
ibility and degradability of a novel injectable-chitosan-based
implant. Biomaterials 23:181–191 (2002).
11 Muller RH and Jacobs C, Buparvaquone mucoadhesive
nanosuspension: preparation, optimisation and long-term
stability. Int J Pharm 237:151–161 (2002).
12 Giunchedi P, Genta, I, Cont, B, Muzzarelli RAA and Conte U,
Preparation and characterization of ampicillin loaded
methylpyrrolidinone chitosan and chitosan microspheres. Bio-
materials 19:157–161 (1998).
13 Lorenzo-Lamosa ML, Remunan-Lopez C, VilaJato JL and
Alonso MJ, Design of microencapsulated chitosan micro-
spheres for colonic drug delivery. Biomaterials 52:109–118
(1998).
14 van der Lubben IM, Verhoef JC and Borchard G, Chitosan and
its derivatives in mucosal drug and vaccine delivery. Eur J
Pharm Sci 14:201–207 (2001).
15 Chandy T and Sharma, CP, Chitosan beads and granules
for oral sustained delivery of nifedipine: in vitro studies.
Biomaterials 13:949–952 (1992).
16 Chandy T and Sharma CP, Chitosan matrix for oral sustained
delivery of ampicillin. Biomaterials 14:949–952 (1993).
17 Banerjee T, Mitra, S, Singh AK, Sharma RK and Maitra A,
Preparation, characterization and biodistribution of ultrafine
chitosan nanoparticles. Int J Pharm 243:93–105 (2002).
18 Gupta KC and Kumar MNVR, Drug release behavior of beads
and microgranules of chitosan. Biomaterials 21:1115–1119
(2000).
19 Janes KA, Calvo P and Alonso MJ, Polysaccharide colloidal
particles as delivery systems for macromolecules. Adv Drug
Deliver Rev 47:83–97 (2001).
20 Ko JA, Park HJ, Hwang SJ, Park JB and Leen JS, Preparation
and characterization of chitosan microparticles intended for
controlled drug delivery. Int J Pharm 249:165–174 (2002).
21 Denuziere A, Ferrier D and Domard A, Chitosan-chondroitin
sulfate and chitosanhyluronate polyelectrolyte complexes.
Physicochemical aspects. Carbohydr Polym 29:317–323
(1996).
22 Hu Y, Jiang XQ, Ding Y, Ge HX, Yuan YY and Yang CZ,
Synthesis and characterization of chitosan poly(acrylic acid)
nanoparticles. Biomaterials 23:3193–3201 (2002).
23 Shiraishi S, Imai T and Otagiri M, Controlled release of
indomethacin by chitosanpolyelectrolyte complex optimiza-
tion and in-vivo in-vitro evaluation. J Control Release
25:217–225 (1993).
24 Sezer AD and Akbuga J, Controlled release of piroxicam from
chitosan beads. Int J Pharm 121:113–116 (1995).
665
CG de Trindade Neto et al
25 Shu X and Zhu KJ, A novel approach to prepare tripolyphos-
phate/chitosan complex beads for controlled release drug
delivery. Int J Pharm 201:51–58 (2000).
26 de Campos AM, Sanchez A and Alonso MJ, Chitosan nanopar-
ticles: a new vehicle for the improvement of the delivery of
drugs to the ocular surface. Application to cyclosporin A. Int
J Pharm 224:159–168 (2001).
27 Aydin Z and Akbuga J, Chitosan beads for the delivery of salmon
calcitonin: preparation and release characteristics. Int J Pharm
131:101–103 (1996).
28 van der Lubben, IM, Verhoef JC, van Aelst AC, Borchard G
and Junginger HE, Chitosan microparticles for oral vaccina-
tion: preparation, characterization and preliminary in vivo
uptake studies in murine Peyer’s patches. Biomaterials
22:687–694 (2001).
29 Berthold A, Cremer K and Kreuter J, Preparation and char-
acterization of chitosan microspheres as drug carrier for
prednisolone sodium phosphate as model for antiinflamma-
tory drugs. J Control Release 39:17–25 (1996).
30 Hejazi R and Amiji M, Stomach-specific antiH pylori therapy.
1. Preparation and characterization of tetracycline-loaded
chitosan microspheres. Int J Pharm 235:87–94 (2002).
31 Bivas-Benita M, Laloup M, Versteyhe S, Dewit J, de Braekeleer
J, Jongert E and Borchard G, Generation of toxoplasma gondii
GRA1 protein and DNA vaccine loaded chitosan particles:
preparation, characterization, and preliminary in vivo studies.
Int J Pharm 266:17–27 (2003).
32 Hara T, Liu F, Liu D and Huang L, Emulsions formulations as
a vector for gene delivery in vitro and in vivo. Adv Drug Deliver
Rev 24:265–271 (1997).
33 Prashanth KVH, Kittur FS and Tharanathan RN, Solid state
structure of chitosan prepared under different N-deacetylating
conditions. Carbohydr Polym 50:27–33 (2002).
34 de Vasconcelos CL, de Medeiros DWO, de Moura KT,
Acchar W, Dantas TNC, Pereira MR and Fonseca JLC,
Polyelectrolyte effect and sedimentation of alumina disper-
sions. Powder Technol 133:164–170 (2003).
666
35 Hare’l G and Ravi GG and Chaim R, Effects of solvent and
agitation on microstructural characteristics of solgel derived
nanocrystalline YTZP powders. Mater Lett 39:63–68 (1999).
36 Pang G, Xu X, Markovich V, Avivi S, Palchik O, Koltypin Y,
Gorodetsky G, Yeshurun Y, Buchkremer HP and Gedanken
A, Preparation of La1−x Srx MnO3 nanoparticles by sonication-
assisted coprecipitation. Mater Res Bull 38:11–16 (2003).
37 Danov KD, On the viscosity of dilute emulsions. J Colloid
Interface Sci 235:144–149 (2001).
38 Buffo RA and Reineccius GA, Modeling the rheology of
concentrated beverage emulsions. J Food Eng 51:267–272
(2002).
39 Cheng NA and Law AWK, Exponential formula for computing
effective viscosity. Powder Technol 129:156–160 (2003).
40 Ngothai Y, Bhattacharya SN and Coopes IH, Estimation of
gelatin layer thickness of polystyrene particles by viscometric
study. J Colloid Interface Sci 193:307–311 (1997).
41 Qian JW, Wang MLHD and Cheng RS, A novel method for esti-
mating unperturbed dimension [η]θ from the measurement
of its [η] in a non-theta solvent. Eur Polym J 37:1403–1407
(2001).
42 Fernandes, ALP, Martins RR, da Trindade Neto CG, Pereira
MR and Fonseca JLC, Characterization of polyelectrolyte
effect in poly(acrylic acid) solutions. J Appl Polym Sci
89:191–196 (2003).
43 Nuessli J, CondePetit B, Trommsdorff UR and Escher F,
influence of starch flavour interactions on rheological
properties of low concentration starch systems. Carbohydr
Polym 28:167–170 (1995).
44 Varum KM, Ottoy MH and Smidsrod O, Acid hydrolysis of
chitosans. Carbohydr Polym 46:287–294 (2001).
45 Zhang X and Bai RB, Mechanisms and kinetics of humic acid
adsorption onto chitosancoated granules. J Colloid Interface
Sci 264:30–38 (2003).
Polym Int 54:659–666 (2005)