Kinetic Study of Chitosan-Tripolyphosphate Complex

Reaction and Acid-Resistive Properties of the

Chitosan-Tripolyphosphate Gel Beads Prepared
by in-Liquid Curing Method

FWU-LONG MI,1 SHIN-SHING SHYU,2 SUNG-TAO LEE,1 TSUNG-BI WONG2

1
Chemistry Division, Department of Physics and Chemistry, Chinese Naval Academy, Taiwan, Republic of China

2
Polymer Division, Department of Chemical Engineering, National Central University, Taiwan, Republic of China

Received 10 April 1998; revised 22 February 1999; accepted 1 March 1999

ABSTRACT: Chitosan gel beads were prepared using an in-liquid curing method by the
ionotropic crosslinking with sodium tripolyphosphate. Crosslinking characteristics of
the chitosan-TPP beads were improved by the modification of in-liquid curing mecha-
nism of the beads in TPP solution. Chitosan gel beads cured in pH value lower than 6
were really ionic-crosslinking controlled, whereas chitosan gel beads cured in pH values
higher than 7 were coacervation-phase inversion controlled accompanied with slightly
ionic-crosslinking dependence. According to the result, significantly increasing the
ionic-crosslinking density of chitosan beads could be achieved by transferring the pH
value of the curing agent, TPP, from basic to acidic. The swelling behavior of various
chitosan beads in acid appeared to depend on the ionic-crosslinking density of the
chitosan-TPP beads that were deeply affected by the curing mechanism of the beads.
The mechanism of chitosan-TPP beads swollen in weak acid was chain-relaxation
controlled, while the mechanism of chitosan-TPP beads swollen in strong acid seem to
be not only chain-relaxation but also chain-scission controlled. Chitosan-TPP beads
prepared in acidic TPP solution decreased the chain-scission ability due to the increase
of ionic crosslinking density of the beads. By the transition of curing mechanism, the
swelling degree of chitosan-TPP beads was depressed, and the disintegration of chi-
tosan-TPP beads would not occur in strong acid. The mechanism of ionic-crosslinking
reaction of chitosan beads could be investigated by an unreacted core model, and the
curing mechanism of the chitosan beads is mainly diffusion controlled when higher
than 5% of chitosan was employed. © 1999 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys
37: 1551–1564, 1999
Keywords: chitosan; tripolyphosphate; ionic crosslinking; in-liquid curing

INTRODUCTION lent bonds to various metal ions by complexation

Chitosan is a copolymer of glucosamine and N-
acetylglucosamine is derived from the natural
polymer chitin. The amine group of the deacety-
lated units of chitosan can form coordinate cova-
Correspondence to: S.-S. Shyu
Journal of Polymer Science: Part B: Polymer Physics, Vol. 37, 1551–1564 (1999)
© 1999 John Wiley & Sons, Inc.
CCC 0887-6266/99/141551-14
in solution. So, chitosan has been a well-known
sorbent of biological origin for metal ions in di-
luted effluents. Porous chitosan beads were ap-
plied to the adsorption of copper, cadmium, chro-
mium, uranium, and so on.1–5 In recent years,
chitosan was already known to be suitable for
biomedical implants, degradable sutures, or
wound dressing, etc.,6 – 8 especially for slow-re-
lease delivery of drugs. In our previous studies,
1551
1552 MI ET AL.
 CHITOSAN-TRIPOLYPHOSPHATE BEADS 1553

chitosan or chitin, having controlled-release char-

acteristics, were prepared to be used as a vehicle
for sustained-release of drugs such as 6-mercap-
topurine, theophylline, or oxytetracycline.9 –11 Be-
sides, it was also used for encapsulation of mac-
romolecules, such as BSA or poly(amino acid)
drugs.12–14
In the attempt to improve material properties
used for metal adsorption or controlled drug-de-
livery, chitosan gel beads was chemically modi-
fied by homogeneous or heterogeneous crosslink-
ing of linear chitosan chains with bifunctional
reagent glutaric dialdehyde (GA) or ethylene gly-
col diglycidyl ether (EGDE).15–21 Crosslinking can
reduce the solubility of chitosan in aqueous sol-
vents over a broad pH range, and increased resis-
tance to chemical degradation or long-term bio-
logical degradation. GA and EGDE are, lately, not
Figure 2. Titration curve: L, NaOH (pH 9.7)/chi-
the preferred crosslinking agents due to their
tosan (0.01 M, pH 4.0); E, TPP (0.01 M, pH 4.0)/chi-
physiological toxicity. It was found that chitosan
tosan (0.01 M, pH 4.0); h, TPP (0.01 M, pH 9.7)/chi-
forms gels with the gentle and nontoxic multiva- tosan (0.01 M, pH 4.0); l, PP(0.0001 M, pH 9.7)/acid
lent counterions, TPP. The ionic interactions be- (pH 4.0); F, TPP (0.001 M, pH 9.7)/acid (pH 4.0); ■, TPP
tween the positively charged amino groups and (0.01 M, pH 9.7)/acid (pH 4.0).
negatively charged counterion, tripolyphosphate,
were used to prepare chitosan beads. The anionic
counterion, TPP, can form either intermolecular
to ionic crosslinking controlled, and higher ionic-
or intramolecular linkages; this is responsible for
crosslinking density of chitosan-TPP beads were
the successful formation of the beads. Bodimeier
produced. Swelling properties of the complex
et al. found that the chitosan-TPP gel beads
beads were analyzed to examine their ionic-
showed pH-dependent swelling and dissolution
crosslinking density. The in-liquid curing mecha-
behavior.13 The beads swelled and dissolved in
nism of chitosan-TPP beads was investigated by
0.1 N HCl, while they stayed intact in neutral
using a heterogeneous unreacted-core model.
fluid. Remunan-Lopez and Bodmeier also re-
FTIR, EDS, and gravity analysis were studied to
ported that chitosan-TPP film was acid nonresis-
confirm the mechanism transition in different pH
tive and would swell to a great extent and finally
values of TPP solutions for the improvement of
dissolve.22 For this reason, the chitosan-TPP bead
crosslinking characteristics of chitosan-TPP
has not been developed to prepare a controlled-
beads.
release dosage for GI tract delivery of drugs due
to its acid nonresistiveness. Besides, the in-liquid
curing mechanism of chitosan-TPP beads was not
EXPERIMENTAL
discussed.
The objective of this study was to improve the
Materials
anti-acid properties of chitosan-TPP beads and
investigate the kinetics of chitosan gel beads pre- Chitosan and sodium tripolyphosphate (TPP)
pared by in-liquid curing method using tri- were purchased from Sigma (USA). Weight-aver-
polyphosphate which has not yet been studied. By age molecular weight (Mw) and deacetylation de-
variation, the pH value of TPP solution from orig- gree are 70,000 g/mol and 87%, respectively. All
inal (pH 8.6) to lower than 6, mechanism of in- other materials were of reagent grade purity. The
liquid curing transferred from a phase-inversion percentage deacylation of the chitosan in this

Figure 1. Ionic reaction of chitosan in TPP aq. solution: (a) deprotonation; (b) ionic
crosslinking.
1554 MI ET AL.
Figure 3. Ladder-loop transition of chitosan-TPP complex structures: (a) ladder type;
(b) loop type.
study was determined from infrared spectral
measurements.23,24
Preparation of Chitosan-Tripolyphosphate
Gel Beads
Chitosan (15 g) dissolved in 500 ml dilute acetic
acid (1% v/v) to prepare the chitosan solution.
Hen egg-white lysozyme (Sigma, USA), 50,000
unit/mg was a commerical product purchased
from Sigma. The lysozyme solution prepared (1.0
ml, 10,000 units/ml) was added to chitosan solu-
tion (19 ml) for digestion, and the mixture (with
500 units/ml of lysozyme) was incubated at 37°C
with mechanical shaking for 2 weeks to decrease
the viscosity. The enzymatic degraded chitosan
solution was lyophilized after thorough dialysis
against deionized water to remove oligomers. Vis-
cosity-average molecular weight (Mw) of enzymic
hydrolyzed chitosan were determined by the
 CHITOSAN-TRIPOLYPHOSPHATE BEADS 1555

Table I. Gravity of Chitosan-TPP Beads (mg/Bead) Analysis of Phosphorus Elution by ICP

Curing The phosphorus elution of various chitosan gel

a
time TPP(8.6) TPP(6.0) TPP(4.0) TPP(2.0) beads were analyzed by dissolving the beads in
hydrochloric acid, diluted with water, and de-
0 min 1.42 1.42 1.42 1.42 tected using a Jarrell-Ash, ICAP 9000 inductively
2 min 1.46 1.46 1.46 1.46 coupled plasma atomic emission spectrometer.
6 min 1.70 1.85 2.20 2.22 The measured concentration of phosphorus in di-
10 min 1.73 1.92 2.46 2.44 luted aqueous was conversed into the phosphorus
15 min 1.79 2.29 2.87 2.86 elution of chitosan beads.
20 min 1.91 2.41 3.13 3.13
30 min 1.96 2.63 3.47 3.43
40 min 2.08 2.79 3.46 3.46 IR Spectra Analysis
60 min 2.09 2.98 3.49 3.47
All chitosan beads cured in TPP solution (various
a
The number in parentheses by TPP is pH. pH values) were analyzed using a Bio-Rad model
FTS-155 spectrophotometer. The peak vibration
of PAO adsorption at 1100 cm21 was detected to
Mark-Houwink equation.25 The viscosity-average monitor the reaction of intermolecular complex
molecular weight of enzymic hydrolyzed chitosan between chitosan and TPP.
was determined to be about 12,000 g/mol. The
finial chitosan solution was dropped into a gently
agitated tripolyphosphate solution (10% w/v) and Electron Scanning Microscopy
stood in the solution for 2 ; 15 min. The pH value The chitosan-TPP gel beads were gold coated to
of tripolyphosphate solutions was adjusted from about 500 3 1028 cm thickness using an Hitachi
original (pH 8.6) to pH 1.0 using 1 M of hydro- coating unit IB-2 coater under a high vaccum, 0.1
chloric acid. The solidified beads were filtered and Torr, high voltage, 1.2 kV, and 50 mA. Coated
washed with 100 ml of deionized water repeat- samples were examined using Hitachi S-2300
edly, then dried in a vacuum oven at 40°C for electron scanning microscopy.
24 h. The final products were stored in a desicca-
tor for future drug release analysis.
EDS (Energy-Dispersive Spectrometer) Analysis
of Phosphorus Distribution
Swelling of Chitosan-TPP Beads
The phosphorus distribution in various chitosan-
The water sorption capacity of chitosan gel beads TPP gel beads was analyzed by EDS (Energy-
was determined by swelling the gel beads in me- Dispersive Spectrometer) analysis. The chitosan-
dia of pH 13 ; 1.2 at room temperature for 12 h, TPP gel beads were carbon coated and then phos-
respectively. A known weight (200 mg) of various phorus distribution was examined using Hitachi
chitosan gel beads were placed in the media for S-2300 electron scanning microscopy with an at-
the required period of time. The wet weight of the tachment of EDS (Delta Class Analyzer, Level I)
swollen beads was determined by first blotting for elemental analysis of phosphorus.
the beads with filter paper to remove adsorbed
water on the surface and then weighed immedi-
ately on an electronic balance. The percentage
RESULTS AND DISCUSSION
swelling of chitosan gel beads in the media were
then calculated from the formula:
The interaction of a strong polycation, chitosan,
with tripolyphosphate (TPP) should result in a
E sw 5 [(W e 2 W o )/W o] 3 100. polycation-multivalent anion complex. The na-
ture and extent of ionic reactions were found to be
where Esw is the percent swelling of gel beads at sensitive to some variable such as the molecular
equilibrium. We denotes the weight of the gel weight, the unit molar ratio, and the charge den-
beads at equilibrium swelling, and Wo is the ini- sity of both electrolytes. Depending on these pa-
tial weight of the gel beads. Each swelling exper- rameters, different types of the chitosan-TPP
iment was repeated three times and the average complexes could be prepared. According to the
value was taken as the percentage swelling value. intrinsic pK (pK0) of chitosan (; 6.3), chitosan
1556 MI ET AL.
Figure 4. IR spectra of chitosan-TPP beads; cured in pH: (a) 1.2; (b) 2.0; (c) 3.0; (d) 4.0;
(e) 6.0; (f) 8.6 of TPP solution.
dissolved in acid to present ONH1 3 site. Tri-
polyphosphoric acid (H5P3O10) is a weak
polyprotic acid-like phosphoric acid, the Ka de-
crease from Ka1 to Ka5 and the decreased quantity
is one to several orders. Due to this reason, so-
dium tripolyphosphate (Na5P3O10) dissolved in
water to dissociate both OH2 and tripolyphospho-
ric ions in the TPP solution. The Kb1 is about 7.45
3 1024 and Kb3 , Kb2 , 1027 , Kb1, this means
that P3O52 42
10 , HP3O10 , and H2P3O10
32
could coexist
in the tripolyphosphate aq. solution in all pH
ranges. In original TPP aq. solution (basic, pH
was not adjusted), the concentration of P3O5210 and
HP3O42 10 was high but the concentration of OH2
was also present. The OH2 or tripolyphosphoric
ions (P3O52 42
10 and HP3O10 ) in this medium could
competitively react ionically with the bind site
ONH1 3 in chitosan by deprotonation or ionic
crosslinking, respectively (Fig. 1). The pH value of
prepared chitosan (0.01M) or TPP aq. solution
(0.01M) is 4.0 and 9.7, respectively. Figure 2
shows the titration curve of chitosan (0.01M) with
NaOH having the same OH2 concentration as
TPP aq. solution (0.01 M). It is shown that even at
the volume mixing ratio of NaOH/chitosan 5 2/1,
the pH value of the mixing solutions stayed at
about 4.0 – 4.1 without change. In this state, the
OH2 ions were almost bound with ONH1 3 of chi-
 CHITOSAN-TRIPOLYPHOSPHATE BEADS 1557

As shown in Figure 3, it seems that a reversible

chitosan-TPP structural change occurs by varying
the pH value of curing agent, TPP. In the higher
pH region, chitosan may take a randomly coiled
conformation because of the decrease of ionized
amino group and of a weak charge repulsion.
However, in the lower pH region, chitosan used
here possibly takes a more extended form both by
hydration of the protonated amino group and by
strong positive charge repulsion between ONH1 3
groups. Thus, in the higher pH range region, the
chitosan-TPP complexes contain several times
more chitosan repeating unit than tripolyphos-
phate expressed as mole ratio and may accept the
looped shape (“loop” means, in this case, TPP
combined with chitosan in a “slackened state”). In
this state, the ionic reaction of chitosan-TPP com-
plex should be a pH-dependent coacervation ac-
Figure 5. Swelling percentage of chitosan-TPP beads companied with slightly ionic-crosslinking.
within 12 h: F, higher crosslinking; E, lower crosslink-
Though according to the facilitation of ionizing
ing.
chitosan in lower pH range region, the chains of
chitosan accept a more extended conformation
and they form a complex with TPP in a ladder-
tosan but not reacted with free hydrogen ions in shaped structure. In this state, chitosan forms
chitosan solution. The pH value of TPP (0.01 M, complex with multivalent counterions, TPP,
pH 9.7)/chitosan (0.01 M, pH 4.0) mixing solution through the formation of intermolecular or in-
increased with the increase of TPP/chitosan mix- tramolecular linkages by ionic interaction. Chi-
ing ratio slowly. This means that the TPP ions tosan–TPP complexes were formed only by the
2
(P3O5210 ) competed with OH ions to bind with ionic interaction between the positively charged
1
ONH3 in chitosan, resulting in the decrease of amino group and negatively charged counterions,
binding sites for OH2 ions and increased pH. TPP ions. The gelation mechanism of chitosan–
When titrating acetic acid (pH 4.0) used TPP TPP complexes in this state should be fully ionic-
(0.01 ; 0.0001 M, pH 9.7), it was found that the crosslinking controlled.
pH of mixing increased quickly. This is due to the
buffer ability of TPP ions in weak acid. However,
the pH increase of the TPP (0.01 M, pH 9.7)/
chitosan (0.01 M, pH 4.0) mixing solution was
significantly slower than the pH increase of TPP
(0.01 M, pH 9.7)/acid (pH 4.0) mixing solution.
This suggests that the TPP ion lost its buffer
ability due to binding with the protonated chi-
tosan. This result shows that the precipitations of
complexes in this state were formed both by dep-
rotonation and ionic crosslinking. By adjusting
the pH value of TPP solution from 9.7 (initial) to
4.0, only P3O52
10 anion ions existed. As can be seen
from Figure 2, the pH value of the TPP (0.01 M,
pH 4.0)/chitosan (0.01 M, pH 4.0) mixing solution
was kept almost constant with the increase of
TPP/chitosan mixing ratio due to no OH2 ions
being released, but chitosan-TPP precipitation
was also formed. This result shows that the precip-
itations of complexes were formed only by ionic- Figure 6. Phosphorus elution of chitosan-TPP beads:
crosslinking between ONH1 3 and TPP ions. ■, higher crosslinking; h, lower crosslinking.
1558 MI ET AL.

Figure 7. Chain-relaxation and chain-scission of chitosan-TPP complexes: (a) slight

chain-scission of higher ionic-crosslinked chitosan-TPP complex; (b) higher chain-scis-
sion of lower ionic-crosslinked chitosan-TPP complex; (c) chain-relaxation of chitosan-
TPP complex due to protonation or hydration.

Chitosan solutions were dropped into TPP
solutions and gelled spheres formed instanta-
neously by ionotropic gelation. Both dissociated
OH2 and tripolyphosphoric ions (P3O52 42
10 , P 3O 10 ,
32)
and H2P3O10 in TPP solution could diffuse
into chitosan droplets during curing in original
TPP solution (pH 8.6). By adjusting the pH
value of the TPP solution from 8.6 (original) to
lower than 7, only tripolyphosphoric ions ex-
isted. Only tripolyphosphoric ions in TPP solu-
tion could diffuse into chitosan droplets to
crosslink with protonated amine group during
curing in acidic TPP solution. Experimental pH
conditions and yields in the preparation of chi-
tosan-TPP complex beads were given in Table I.
By the variation of pH value of TPP solution
from acidic to basic, this significantly increased
the weight of chitosan-TPP beads due to the
increase of linked TPP ions within the chitosan-
TPP beads (Table I). The chitosan-TPP beads
prepared by curing in different TPP solutions
were spectral analyzed by FT–IR. As described
previously, the extent of ionic-crosslinking den-
sity was significantly dependent on the pH

Figure 7. (Continued from previous page)
value of TPP solution. Figure 4 showed the IR
spectra of chitosan-TPP beads prepared in differ-
ent pH values of TPP solution. The presence of
PAO group is indicated by the peak at the fre-
quency of 1150 cm21. By keeping the curing time
at 30 min, the intensity of PAO absorbance in-
creased along with the decrease in pH values of
the curing agent, TPP solution, which corre-
sponds to the fact that sample weights of chitosan
beads decreased with increasing the pH value of
CHITOSAN-TRIPOLYPHOSPHATE BEADS 1559
the TPP solution. The increase of the intensity of
PAO absorbance indicated the increase of bound
TPP ions. This result would be attributed to the
increase of interchain linkage of ONH13 groups in
chitosan by TPP ions. It demonstrated that the
ionic reaction of chitosan-TPP beads were signif-
icantly influenced by the pH value of TPP solu-
tion, and the ionic-crosslinking density could be
improved by the variation of the pH value of the
curing agent, TPP solution.
1560 MI ET AL.
Figure 7. (Continued from previous page)
A convenient proof of crosslinking is the swell-
ing behavior of the chitosan-TPP beads in various
pH values of aqueous media. Swelling ability of
various chitosan beads were carried out in disso-
lution media of pH 1.2 ; 14. Figure 5 shows the
equilibrium swelling behavior of two kinds of chi-
tosan-TPP beads synthesized from different bind-
ing ratios of TPP to chitosan (cured in original
TPP or acidic TPP aq. solution). It was indicated
that swelling ability of the chitosan-TPP beads
was quite different according to different pH val-
ues of the dissolution medium. In pH 3 ; 7 media,
the ionic-crosslinked chain of chitosan-TPP beads
didn’t dissociate, so the swelling of chitosan-TPP
beads in this medium would be mainly attributed
to the hydration or ionization of unbound ONH2
sites in chitosan. Due to this reason, the swelling
percent of various chitosan-TPP beads in this me-
dia were very low (lower than 60%) for both
higher and lower ionic-crosslinked chitosan-TPP
 CHITOSAN-TRIPOLYPHOSPHATE BEADS 1561

ionic crosslinking density and the anti-acid charac-

Figure 8. Effect of ionic strength on equilibrium
swelling degree of chitosan-TPP beads.
teristic of the chitosan-TPP beads could be im-
proved by the modification of the in-liquid curing
mechanism of the chitosan-TPP beads. Figure 8
shows plots of equilibrium degree of swelling vs.
ionic strength for the chitosan-TPP gel beads in pH
1.2, NaCl aq. solution. For the lower ionic-
crosslinked chitosan-TPP beads (gelled in original
basic TPP aq. solution), the equilibrium swelling
degree stays constant until the contents of NaCl
(aq.) reaches 0.005 M, and then decreases with fur-
ther increase in concentration of NaCl (aq). This
induces deswelling of the gel beads in a medium of
high ionic strength due to the binding of anionic ion
(Cl2) to protonated ONH1 3 site of chitosan.
Figure 9 shows the EDS analysis of the line
profiles of phosphorus distribution on the cross-
section of various chitosan-TPP beads which were
prepared in different pH values of TPP solution.

beads (gel beads cured in original TPP or acidic

TPP aq. solution). In this pH range, swelling of
the chitosan-TPP beads was mainly attributed to
chain-relaxation of chitosan-TPP complex by the
protonation or hydration of the unbound ONH2
groups but not by the scission of ionic-crosslinked
chain. When the pH value of dissolution medium
was decreased from pH 3 to pH 1–2, the swelling
of chitosan beads were increased to a different
degree depending on the in-liquid curing process
of the gel beads. In this pH range, the chitosan
beads prepared by curing in different TPP solu-
tions displayed different swelling characteristics
due to the variation of ionic-crosslinking density
of the beads. In pH 1 ; 2 of dissolution media, the
chitosan gel beads prepared by curing in original
TPP solution swelled quickly and gradually dis-
solved within 12 h, whereas chitosan gel beads
prepared in acidic TPP solution only very slightly
swelled but were not dissolved in this media (Fig.
5). Figure 6 showed the phosphorus elution of
chitosan-TPP beads in pH 1.2 ; 6 media. This
result indicated that the chitosan gel beads pre-
pared in acidic TPP solution had a lower degree of
phosphorus chain-scission than that of chitosan
gel beads prepared in original TPP solution. This
would be attributed to the more stable structure
of chitosan-TPP complexes due to the high degree
of interchain linkages in them. The schematic
representation of chain-relaxation or chain-scis- Figure 9. EDS analysis of the line profiles of phos-
sion of the chitosan-TPP complexes was shown in phorus distribution in chitosan-TPP beads: (a) cured in
Figure 7. This result confirms our theory that the pH 8.6 TPP; (b) cured in pH 2.0 TPP.
1562 MI ET AL.
Figure 10. In-liquid curing mechanism of chitosan-TPP beads: (a) coacervation; (b)
ionic-crosslinking.
The line profile of phosphorus on the cross-section
of chitosan beads gelled in original TPP solution
(pH 8.6) has no significant variation with the
radial direction. Whereas, signal of phosphorus
decreased with the decreasing of radial distance
away from core for chitosan beads gelled in acidic
TPP solution (pH 2.0). Besides, signal signifi-
cantly increased with increasing the gelling time
of chitosan-TPP beads prepared in this acidic TPP
solution. This result as well as the FT–IR studies
suggested that the ionic interaction between chi-
tosan and negatively charged counterion, tri-
polyphosphate (TPP) was dependent on the pH
value of the TPP solution. According to the line
profiles of phosphorus distribution in chitosan-
TPP beads, the in-liquid curing mechanism of
chitosan-TPP beads could approach a heteroge-
neous fluid–solid reaction of an unreacted core
model but not a progressive conversion model. In
the original TPP solution, the OH2 ions competed
with P3O5210 ions to react with the protonated
amino group of chitosan on the surface of beads as
soon as the chitosan droplets contact with the
TPP solution. After the formation of a gelled outer
layer, the resistance for the larger P3O52
10 ions to
diffuse through the gelled film into the inside

matrix was higher than the resistance of OH2
ions to diffuse into the beads, due to the smaller
molecular size of OH2. So, the gelled mechanism
of chitosan beads was mainly dependent on coac-
ervation-phase inversion due to the neutraliza-
tion of protonated amino groups of chitosan ac-
companied with slightly ionic crosslinking [Fig.
10(a)]. Whereas, there were no OH2 ions in TPP
solution at pH values lower than 7. The gelation
of chitosan beads was almost controlled by ionic
crosslinking throughout the beads due to the for-
mation of intermolecular or intramolecular link-
ages between linear chitosan chain by multiva-
lent anion, TPP [Fig. 10(b)]. According to this
theory in our study, the crosslinking density of
chitosan-TPP gel beads could be improved by the
modification of the in-liquid curing mechanism.
The kinetic mechanism of chitosan-TPP beads
gelling in TPP solution was gravity investigated
by a heterogeneous fluid–solid reaction, unre-
acted-core model. The reactions occur first at the
outer skin of the gel bead, the zone of reaction
then moves into the unreacted core and may leave
behind completely converted material. In this
process, penetration and diffusion of P3O52 10
through the gelled layer to the surface of the
unreacted core and reaction on the surface would
be the rate-controlling factors. Because the for-
mation of ionic-crosslinked layers on chitosan
droplets was homogeneous and the formed
crosslinked layer was dense enough, resistance
for diffusion of P3O5210 through the film usually
became very large. The in-liquid curing mecha-
nism of chitosan-TPP gel beads prepared by the
ionic-crosslinking (cured in acidic TPP solution)
process would be diffusion through ionic-
crosslinked layer control. The equation of the
model is expressed as
t/ t 5 1 2 3~1 2 X! 2/3 1 2~1 2 X! (1)
t 5 r R 2/6D e C AO (2)
where X denotes the ionic-crosslinking conversion
fraction of chitosan beads at time t, t is the time
for complete reaction, CAO is the bulk concentra-
tion of P3O52
10 , De is the effective diffusion coeffi-
10 in the crosslinked layer, r is the
cient of P3O52
molar density of chitosan in particle, and R is the
radius of the spherical particles.
This model shows that the conversion of chi-
tosan by phosphorus ionic-crosslinking reaction
was dependent on the reaction time (t) and the
CHITOSAN-TRIPOLYPHOSPHATE BEADS 1563
Figure 11. Unconversion fraction (1-X) of chitosan-
TPP beads vs. time (min): F, cured in pH 2.0 TPP,
chitosan (5% w/v); E, cured in pH 2.0 TPP, chitosan (3%
w/v); ■, cured in pH 8.6 TPP, chitosan (5%).
initial concentration of crosslinking agent (CAO).
Figure 11 shows the curve of conversion fraction
of chitosan beads vs. time by curing in pH 2.0 of
TPP solution. By fitting the data of conversion
fraction (1-X) and reaction time (t) to eq. (1), the
kinetics of the heterogeneous curing mechanism
could be examined. It was observed that the cur-
ing mechanism of beads was diffusion of TPP ions
into spherical particles. A significant approach to
diffusion-controlled mechanisms was observed by
the large increase in the concentration of chi-
tosan. The result indicated that the resistance for
P3O5210 to diffuse through the ionic-crosslinked
layer to react on the surface of the unreacted core
increased significantly due to increasing the den-
sity of crosslinked layers. Kinetics of the ionic-
crosslinking of chitosan-TPP beads could be well
fitted with the diffusion-controlled mechanism of
unreacted core models when higher than 5% w/v
of chitosan was employed.
Due to the formation of complex gel layers in a
nonhomogeneous matrix which contained coacer-
vation and ionic-crosslinking blocks, the effective
diffusion coefficient De of P3O52
10 in the crosslinked
layer was not kept constant for chitosan-TPP
beads prepared by curing in basic TPP solution
(phase-inversion mechanism accompanied with
slightly ionic-crosslinking). It is observed that the
curing mechanism of chitosan-TPP beads pre-
pared in this state was deviated from the diffu-
sion-controlled unreacted-core model. This result
1564 MI ET AL.
indicated that the in-liquid curing mechanism of
chitosan-TPP gel beads prepared by phase-inver-
sion accompanied with slightly ionic-crosslinking
(cured in basic TPP solution) processes may be
controlled or influenced by other mechanisms.
CONCLUSION
The in-liquid curing mechanism of chitosan beads
gelling in TPP solution could be easily modified
from pH-dependent coacervation to ionic crosslink-
ing reaction only by adjusting the pH value of TPP
solution. The swelling behavior of various chitosan
beads in acid appeared to depend on the in-liquid
curing mechanism of the beads. Higher ionic-
crosslinking density of chitosan beads has less
swelling ability due to their high stabiltiy in acid. In
this study, the ionic-crosslinking density could be
controlled by modifying the in-liquid curing mecha-
nism and the material properties of the chitosan gel
beads might be improved. Besides, the mechanism
of ionic-crosslinking of chitosan-TPP beads was also
investigated by a heterogeneous fluid–solid reac-
tion, unreacted core model.
The authors wish to thank the National Science Coun-
cil of the Republic of China for financial support (NSC
85-2331-B-008-001-M08).
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