World Journal of Microbiology & Biotechnology (2005) 21: 913920 DOI 10.

1007/s11274-004-6559-5

Springer 2005

Chitosan interactions with metal ions and dyes: dissolved-state vs. solid-state application
E. Guibal*, E. Touraud and J. Roussy Ecole des Mines dAle`s, Laboratoire Genie de lEnvironnement Industriel 6, avenue de Clavie`res, F-30319 ALES cedex, France *Author for correspondence: Tel. +33-0-466782734, Fax: +33-0-466782701, E-mail: Eric.Guibal@ema.fr
Received 13 October 2004; accepted 22 November 2004

Keywords: Adsorption, chitosan, coagulationocculation, mercury, reactive black 5, ultraltration

Summary Chitosan is an amino-polysaccharide with highly efcient properties for the binding of metal ions and anionic dyes. Uptake may occur through chelation on free amino functions (at near-neutral pH) or by electrostatic attraction on protonated amino groups (in acidic solutions). The polymer is soluble in acidic solutions and its binding properties can be used in both solid form (sorption) and liquid form (ultraltration coupled with chelation, coagulation occulation). These properties have been used for the recovery of mercury from dilute solutions at initial pH 5 (which reveals the most efcient pH in the range pH 46) and for the recovery of Reactive Black 5 (RB5, anionic dye) at pH 3. While in the case of mercury binding saturation of the biopolymer is only slightly higher when chitosan is used in the liquid form compared to solid-state adsorption, in the case of the coagulationocculation of RB5 (using the liquid-form of chitosan) the saturation of the polymer (calculated on the basis of molar ratio of dye vs. amino groups of the polymer) is reached at a signicantly greater value than when the polymer is used for the solid-state binding of the dye. There is a much more efcient use of amino groups when chitosan is used in the liquid-form due to a better availability of amino groups (less hydrogen bonds between the chains of the polymer) and to a better accessibility to internal sorption sites (lower diffusion control).

Introduction The increasing demand for new processes for wastewater treatment, more environmentally friendly and more competitive, has led to much research on the use of biopolymers such as alginate and chitin/chitosan for the recovery of metal ions (Chen & Wang 2001; Guibal 2004) or organic compounds (Juang et al. 1997; Annadurai et al. 2002; Chiou & Li 2002; McCarrick et al. 2003). These biopolymers are characterized by high sorption capacities, easy degradation routes (at the end of life cycle) and relatively low costs (compared to sophisticated resins, Dubois et al. 1995). These properties make them very promising for replacing conventional processes. Chitosan has a unique property among these biopolymers: its cationicity in acidic solutions. This property is due to the presence of a large number of amino groups, which are very reactive for (a) metal cations by chelation in near-neutral solutions, and for (b) metal anions that can be bound to protonated amino groups in acidic solutions (Guibal 2004). The pKa of the amino groups of chitosan strongly depends on the degree of acetylation of the polymer and charge neutralization in solution (Sorlier et al. 2001). The protonation of amino groups also involves the dissolving of the

biopolymer in acidic solutions (with the exception of dilute sulphuric acid solutions). For some applications involving metal sorption at low pH the polymer can be cross-linked to prevent its dissolving, but at the expense of a decrease of the number of free amino groups. This parameter is very limiting in the case of metal cation chelation (Dzul Erosa et al. 2001), but less important in the case of electrostatic attraction mechanisms (Ruiz et al. 2000). Alternatively, it is possible to use chitosan without cross-linking treatment but, at the saturation of the polymer, it is necessary to recover the metal-bound macromolecules by an ultraltration process. This is the base of polymer-enhanced ultraltration processes (PEUF) (Juang & Tseng 2000; Kuncoro et al. 2004). The protonation of chitosan in acidic solution may be also interesting for the electrostatic attraction of anionic dyes (McKay et al. 1987; Yoshida & Takemori 1997; Annadurai et al. 2002; Gibbs et al. 2003, 2004; Wong et al. 2004). These chitosan-dye interactions are very interesting for the sorption of dyes on solid chitosan (using sulphuric acid for pH control) but they can be also used for dye binding using chitosan in a dissolvedstate. In this case, the interactions of the dyes with chitosan result in the formation of mixed colloids (dye/ chitosan) that can settle after a long contact time or can

914 be recovered (after maturation) by ltration. Protonated amino groups neutralize anionic charges of the dye (coagulation eect), before neutralized dyes aggregate and settle (occulating eect). This property results in coagulationocculation properties that have been used elsewhere for the treatment of many groundwaters and industrial euents (from the agricultural and food industries) (Huang & Chen 1996; Pan et al. 1999; Huang et al. 2000; Roussy et al. 2004). The present work focuses on (a) the study of mercury recovery on chitosan by sorption (chitosan in solidstate) and PEUF (chitosan in dissolved-state), and (b) the study of Reactive Black 5 (RB5, or Intracron Black) recovery on chitosan by sorption and by coagulation occulation. The main objective of this work is thus to demonstrate that, despite the high efciency and easy use of chitosan for sorption processes (in the solidstate), the use of chitosan in the dissolved-state allows improving the reactivity of the polymer. This improvement in uptake properties will be measured through equilibrium performance but also kinetic considerations.

E. Guibal et al. material and an inert product: the actual true dye content is 55%. This must be taken into account for the evaluation of true molar ratio and true sorption capacities (concentrations). The experiments have been performed using the commercial salt without purication and the concentrations are given in function of the total amount of commercial salt used for the preparation of the solutions. However, for the evaluation of the molar ratio between the dye and amino groups of chitosan, the correction was made taking into account the purity of the salt, as well as the humidity and the degree of acetylation of chitosan. Mercury (in the form of nitrate salt) was supplied by Fluka AG (Switzerland). Other common reagents (acids, bases) were supplied by Fluka (Switzerland) and Carlo Erba (Italy) as analytical grade products. Sorption Processes

Sorption isotherms were performed in batch systems using a standard procedure (Procedure 1) consisting in the mixing of a xed volume of solution (at given concentrations) with different amounts of chitosan for at least 48 h (depending on particle size). In the case of Material and methods mercury two different procedures have been obtained: (a) the previously described method (Procedure 1); and Material (b) an alternative procedure consisting in mixing a xed amount of sorbent with a xed volume of solution Chitosan was used as supplied by Aber Technologies prepared at concentrations ranging between 10 and (Plouvien, France). The material was previously 100 mg Hg l)1 (Procedure 2). The initial pH was concharacterized by the FT-IR technique and SEC trolled with sulphuric acid and NaOH for experiments (coupled with refractometer and laser light scattering on both mercury and RB5. Solutions were ltered at the measurements) for the determination of the degree end of the experiment using a 1.2 lm Whatman ltraof deacetylation and polymer weight, which were tion membrane. The residual concentration was mea87% and 125,000 g mol)1, respectively. Chitosan sured by Inductively Coupled Plasma Atomic Emission was ground and sieved in 4 size fractions: Spectrometry (ICP-AES) (JY 2000, Jobin-Yvon, G1 < 125 lm < G2 < 250 lm < G3 < 500 lm < G4 France) for mercury concentration and by u.v.visible < 710 lm. For the experiments using chitosan in spectrophotometry for dye concentration at the wavedissolved-state, the polymer was dissolved in acetic length of 598 nm (UVVIS spectrophotometer Shimaacid solution (1 g of chitosan for 1 g of acetic acid, dzu UV-160 A). A drop of nitric acid (10%) was added to mercury solution before ICP-AES analysis, while a 80% (w/w)). Reactive Black 5 (RB5) was supplied by Aldrich (USA). drop of sodium hydroxide (5 M) was added to dye This dye is characterized as a diazo compound bearing 4 samples to maintain a constant pH of the samples before sulphonyl groups (Figure 1), Mr 991.82 g mol)1. The analysis (indeed, a change in the pH of the sample may commercial salt is supplied as a mixture of the active cause a change in the dyes u.v.visible spectrum and may aect the determination of solution absorbance and dye concentration). The mass balance equation was used for determining solute concentration on the sorbent at equilibrium. The pH of the solution at equilibrium was O systematically measured using a WTW 526 pH-metre NaO3SOCH2CH2 S N=N SO 3Na (WTW, Germany): the pH measurements were performed with a 0.05 pH-unit precision. HO Sorption kinetics were performed by mixing 1 l of Reactive Black 5 solution at xed concentrations (either mercury salt or dye) with xed amounts of sorbent. The solution was 2HN O agitated for at least 48 h and samples were regularly collected, ltered and analysed for the determination of NaO3SOCH2CH2 S N=N SO3Na residual solute concentration. Ultraltration experiments were performed in batch Figure 1. Structure of Reactive Black 5. system using an Amicon 8400 ultraltration unit with

= =
O

= =
O

Chitosan interactions with mercury and dyes polyethersulphone membranes (Cut-off: 50 and 100 kDa). The pressure was xed at 2 bars. Typically, the solution containing the polymer (dissolved in HCl) was mixed with mercury solution at xed pH for 1 h and nally the solution was permeated though the cell. The molar ratio Hg/polymer was varied in order to evaluate polymer saturation. Permeate samples were regularly collected and analysed for metal determination (ICP/AES) and polymer content (TOC metre, Shimadzu, Japan). Experiments on dye coagulation and occulation were performed using a jar-test equipment. The dye solution, which pH was controlled using HCl, was mixed after polymer addition under strong agitation (ca. 200 rev/ min) for 3 min and then under slow agitation (ca. 40 rev/min) for 20 min. After stopping the agitation, samples were regularly collected and analysed for dye content in the supernatant. Two different experimental procedures have been used depending on the experiments. The amount of dye on the polymer was calculated by a mass balance equation assuming that the amount of dye that was removed from the solution was bound to the total amount of polymer added in the solution. The dye/polymer molar ratio was varied in order to determine the optimum relationship between dye functional groups and amino groups of chitosan. The optimization took into account both the removal efciency and the molar ratio dye/amino groups.

915 different experimental procedures for initial mercury concentrations below 50 mg l)1. The pH was not controlled during the experiment; it was only monitored at the end of the experiment. The variation of the pH during sorption experiments is an experimental parameter frequently underestimated in the literature dedicated to biosorption properties of chitosan; for this reason special attention has been paid to pH variation in the present work. The change in the pH in the course of metal sorption was directly inuenced by experimental conditions; i.e. initial pH, amount of sorbent and initial metal concentration (Figure 3). When the initial pH of the solution approaches the pKa of chitosan, which depends on the degree of deacetylation and the extent of amino groups dissociation (Sorlier et al. 2001), the change in the pH decreased (Figure 3a). This weak pH variation may be explained by a buering eect of chitosan on the solution: the protonation/deprotonation of amino groups tends to decrease and the pH is less aected. At low initial pH (i.e. pH 4) the change in the pH increased with initial metal concentration, this trend was reversed at higher initial pH (Figure 3b). No explanation was found to this increased eect of initial metal concentration on pH variation at pH 4. At pH 5 and 6, increasing the amount of metal resulted in an increase of the amount of metal adsorbed and therefore a reduced number of amino groups available for the uptake of protons (additionally to the buering eect of chitosan when approaching the pKa of amino groups of chitosan). When initial pH was 4 the pH variation reached 0.8 pH unit (as a mean value), while when the initial pHs

Results and discussion Chitosan interactions with mercury ions Adsorption process The sorption process on chitosan is strongly controlled by the pH of the solution due to protonation of amino groups on the biopolymer. This may have a positive effect on the uptake of anionic solutes or a negative impact for example in the case of cationic molecules. In acidic solutions, the protons compete with target cations and signicantly reduce their sorption. Figure 2 shows the sorption isotherms obtained at pH 46 using 2
500 400 q (mg Hg/g) 300 200 100 0 0 10 20 30 Ceq (mg Hg/L) 40
pH 4 pH 4 pH 5 pH 5 pH 6 pH 6

(a) 7
mchitosan: 25 mg pHi 4 pHi 5

Final pH

5
Vsol: 150 mL

pHi 6

4 0 25 50 75 100 Initial Hg concentration (mg/L)

(b) 7.5
pHi: 4; Co: 10 mg/L pHi: 4; Co: 25 mg/L pHi: 4; Co: 50 mg/L pHi: 5; Co: 10 mg/L pHi: 5; Co: 25 mg/L pHi: 5; Co: 50 mg/L pHi: 6; Co: 10 mg/L pHi: 6; Co: 25 mg/L pHi: 6; Co: 50 mg/L

6.5 Final pH 5.5 4.5 3.5 0 0.02 0.04 0.06

Vso l: 150 mL

0.08

Chitosan amount (mg)

Figure 3. Inuence of mercury concentration (a) (Procedure 1) and chitosan amounts (b) (Procedure 2) on equilibrium pH (pHi: initial pH, and C0: initial metal concentration).

Figure 2. Inuence of pH on mercury sorption on chitosan akes (closed symbols: Procedure 1; open symbols: Procedure 2).

916 were 5 and 6 the pH variation tended to 0.5 and 0.3 pH unit, respectively. Figure 2 shows that the pH hardly changed the prole of the sorption isotherm in the rst part of the curve: this weak effect is due to this buffering effect of the polymer on the nal pH; the polymer smoothed the differences in the initial pHs. Figure 4 shows a more complete isotherm obtained at pH 5 with different particle sizes (G1 < 125 lm < G2 < 250 lm < G3 < 500 lm). At equilibrium, when a sucient time of contact is maintained between the sorbent and the solution, the sorption performance was not drastically changed when increasing the size of sorbent particles. This means that all the available sites (in the whole mass of the sorbent) can be accessed, providing a sucient contact time. The pH variation for each experimental series has been monitored (not shown). At initial pH 5, the pH variation increased (a) at low metal concentration, (b) with increasing amounts of sorbent, and (c) very signicantly when the size of sorbent particles decreased. The maximum sorption capacity tended to 520 mg Hg g)1. This means a sorption capacity close to 2.5 mmol Hg g)1. Taking into account the intrinsic characteristics of the chitosan sample used in this study (i.e. humidity or water content: 10%; deacetylation degree 87%, resulting in a molecular weight of the equivalent monomer unit: 166 g mol)1) it is possible to convert this maximum sorption capacity in molar units (mol Hg/mol R-NH2). The saturation ratio gives 0.53 mol Hg per mole of free amino groups. Figure 5 shows the kinetic proles obtained for given experimental conditions (i.e. initial metal concentration: 50 mg Hg l)1; sorbent dosage: 200 mg l)1) for 72 h of contact time. A greater contact time appears necessary since the nal concentrations were dierent (increasing with the size of sorbent particles) contrary to the results obtained from Figure 4 (in this case the contact time was set at 5 days). The time required to reach the equilibrium signicantly increased with the size of sorbent particles: from 8 h for small particles (below 125 lm) up to 48 h for large particles. This means that intraparticle diusion greatly inuences the accessibility of metal ions to internal sites. These diusion restrictions may have a critical impact on the design of
1 0.8 C(t)/Co 0.6 0.4 0.2 0 0 24 Time (h) 48
Co: 50 mg Hg/L SD: 200 mg/L

E. Guibal et al.

G1 G2 G3

72

Figure 5. Inuence of particle size on the kinetics of mercury sorption (initial pH 5).

operating plant. Decreasing the size of sorbent particles for improving kinetic behaviour results in strong impact on head loss and column blockage (when the sorbent is to be used in xed-bed systems). Polymer-enhanced ultraltration process In this case, to maintain chitosan soluble, HCl was used instead of sulphuric acid. This is basically the only difference between the experimental conditions relative to solution preparation. The presence of chloride may affect the speciation of mercury in solution. It has been shown that mercury can form chloro-species that can be adsorbed on protonated amino groups in acidic solutions (Kawamura et al. 1993). However, in the present case, most experiments were performed at low acidity (pH 55.5) and the contribution of chloride to the change in metal speciation is not expected to really aect uptake performance. Figure 6 shows the impact of pH on the recovery of mercury in presence of chitosan. As expected, increasing the pH improved the eciency of metal recovery. The increase in the pH contributes to the deprotonation of amino groups that in turn bind mercury ions by chelation; the macromolecule loaded with metal ions is therefore retained by the membrane. At low pH (i.e. pH 2), a partial increase in metal retention was observed, presumably due to the binding
100 98 100 80 RHg (%) 60 40 20 0 1 2 3 4 pH 5 6 7

600 q (mg Hg/g)

RP (%)
G1 G2 G3

96 94 92 90

400

200

0 0 50 100 Ceq (mg/L) 150

Figure 4. Inuence of sorbent particle size on mercury sorption at initial pH 5.

Figure 6. Inuence of pH on mercury retention by ultraltration using chitosan as the microligand (RP: polymer retention; RHg: mercury retention; membrane cut-o: 50 kDa; polymer concentration: 200 mg l)1; mercury concentration: 25 mg Hg l)1).

Chitosan interactions with mercury and dyes of chloro-mercuric species on protonated amino groups. The molar ratio between metal and polymer has been varied in order to evaluate metal retention (Figure 7). The mass balance equation was used for calculating the molar ratio between mercury ions and the amino groups on the biopolymer. The hypotheses used for the calculation of this molar ratio follow: (a) the polymer is considered to be completely retained by the membrane; (b) the metal retained by the polymer is deduced from mercury concentration in the ultraltrate. The humidity of raw chitosan and the deacetylation percentage are taken into account to obtain the pseudo binding isotherm that represents the binding capacity q (molar ratio: mol Hg/mol R-NH 1 ) in function of mercury 2 concentration in the ultraltrate (Figure 7). The amount of mercury in the retentate is obtained by a mass balance between the initial amount of mercury and the amount of mercury present in the ultraltrate; compared to the amount of polymer used (and retained in the retentate) the ratio gives the binding capacity. This curve shows the typical shape of the Langmuir equation with an asymptotic trend. The concentration range (below 0.4 mmol Hg l)1, i.e. below 80 mg Hg l)1) was shorter than for sorption isotherms (Figure 4). In this concentration range the curve did not reach the saturation plateau that should exceed 0.5 mol Hg mol R-NH . 2 Anyway, in the case of mercury uptake the use of chitosan in a dissolved-state did not much change the binding capacity compared to solid-state adsorption. This did not greatly increase the availability of amino groups. However, the kinetics of the process were greatly improved. Indeed, in the case of solid-state use

917 of chitosan the time required for reaching the equilibrium exceeded several hours (Figure 6), in the case of PEUF, the time required for metal separation was greatly decreased. The time used for complexation the dissolved state and pH control, and stabilization was approximately set experimentally at 1 h. The dissolved state makes the amino groups much more accessible to metal ions: the polymer network is opened; the hydrogen bonds between polymer chains in the solidstate break during the dissolving step, these active sites are more available and easier to diuse to. Chitosan interactions with RB5 Adsorption process Though the acid base properties of this dye were not recorded, sulphonyl groups usually have pKa below 4. Anionic dyes are adsorbed by electrostatic attraction on protonated amino groups of chitosan in acidic solutions. Experiments have been performed at pH 3. Results are summarized on Figures 8 and 9 for sorption isotherms and uptake kinetics respectively. The sorption isotherm was characterized by a very favourable shape: the initial slope of the curve was very steep (indicating a great anity of the sorbent for the solute) and the plateau of the Langmuir-type isotherm was reached for a very low residual concentration (close to 5 mg l)1). The results were modeled using the Langmuir equation with a good correlation coecient. The maximum sorption capacity was closed to 1100 mg dye g)1. According to previous comments, the molar ratio between the dye and the amino groups of the biopolymer at saturation tends to 0.13 mol of dye per mole of amino group; this means more than 7 mol of amino groups for the sorption of 1 mol of dye (i.e. more than 7 mol of amino groups for 4 mol of sulphonyl groups). This greatly exceeds the stoichiometry required for the neutralization of charges of sulphonyl groups. This dierence clearly demonstrates that all the amino groups are not really active/ available or at least accessible. The hydrogen bonds linked between monomer units of the same chain (intramolecular bonds) or between monomer units of dierent chains (intermolecular bonds) decrease their reactivity. The weakly porous structure of the polymer

(a) 100

80 RHg (%) 60 40 20 0 0
(b) 600

10 15 20 L/M (mol/mol)

25

1200 900 q (mg/g) 600 300

Reactive Black 5

q (mmol Hg/mol)

400 200 0 0 0.1 0.2 0.3 Ceq (mmol Hg/L) 0.4

pH 3

0 0 50 100 150 Ceq (mg/L) 200

Figure 7. Inuence of Ligand/Metal molar ratio on metal retention at initial pH 5 (a), and binding isotherm for mercury using chitosan as the macroligand at initial pH 5 (b).

Figure 8. Reactive Black 5 sorption isotherm at pH 3 using chitosan akes (G1).

918
(a) 1
0-63 m ReactiveBlack 5 Co: 100mg/L SD: 150 mg/L pH3 63-125 m 125-180 m

E. Guibal et al.
(b) 1

0.8 C(t)/Co 0.6 0.4 0.2 0 0

0.8 C(t)/Co 0.6 0.4 0.2 0

180-250 m 250-355 m 355-500 m

12

24 Time (h)

36

48

3 4 Time (h)

Figure 9. Inuence of particle size on Reactive Black 5 sorption kinetics at initial pH 3 ((a): complete kinetics, (b): initial stage).

and its residual crystallinity are also critical parameters for the hydration and the accessibility to sorption sites as evidenced by Piron et al. (1997). In the solid-state, the large dye molecules have increasing diculty to penetrate porous network, steric hindrance being increased with the progressive saturation of the sorbent. Figure 9 shows the kinetic proles for the sorption of RB5 using 6 dierent sizes of chitosan particles. This gure clearly demonstrates that the recovery of the dye was controlled by intraparticle diusion: the time required for reaching equilibrium dramatically increases when increasing the size of sorbent particles. For the largest particles, even after 48 h of contact the equilibrium was not reached. The impact of particle size was much greater than in the case of mercury sorption: this may be explained by the dierence in the size of solute molecules. The diusivity of the solute in the solid had been shown to be a function of the ionic radius of solute molecule to the pore radius of solid particles (Krajewska 2001). Even for the smallest particles, 23 h of contact times were required for achieving the complete recovery of the dye (or at least for reaching the equilibrium). Coagulation and occulation process The mechanism of coagulation can be explained by a charge neutralization mechanism. The interactions of the dye (bearing several sulphonyl groups) with different monomer units of the polymer (on the same chain or on vicinal chains) leads to a kind of cross-linking effect that induces the gelation of the polymer chains and the insolubilization of the material (formation of ocs). This is a mechanism similar to that observed in the case of ionotropic gelation (Mi et al. 1999; Dambies et al. 2001). Figure 10 shows the relative abatement of the absorbance of dye solutions collected at the top of sedimentation tanks after ltration in function of chitosan concentration and initial dye concentration. Each of these curves were characterized by a two-line plot: a rst steep decrease to reach a minimum of absorbance, followed by another steep increasing line (with a higher steepness); in some cases the curve was followed by another line with a weak slope. The slope of the rst section of the curves tended to decrease with increasing dye concentration, while that of the second section of the curves remained almost constant. These

1 0.8 C/Co 0.6 0.4 0.2 0 0 10 20 30 40 50 Chitosan concentration (mg/L) 60

Co: 100 mg/L Reactive Black 5 pH 3

Co: 25 mg/L

Co: 50 mg/L

Figure 10. Inuence of chitosan concentration on Reactive Black 5 removal by coagulation occulation at dierent dye concentrations (2550100 mg l)1) at initial pH 3 (bold, grey and opened symbols and bold, grey and dashed lines show same experimental series, the dierence in the representation style is only used to materialize the change in the slope of the curves).

curves may be explained by the destabilization of the dye solution followed by a re-stabilization of the solution resulting from the addition of an excess of chitosan in the solution. In the rst part of the curves, the protonated amino groups introduced in dye solution allowed the charge neutralization of anionic sulphonyl groups. However, when the amount of chitosan added to the solution exceeded the amount required for their neutralization, the global charge of the dye-chitosan complexes turned to a positive value, which in turn induced the repulsion of these positively charged molecules. These curves delimited a minimum chitosan concentration for optimum colour abatement. This concentration logically increased with the concentration of the dye: 5, 15 and 35 for dye concentrations of 25, 50 and 100 mg l)1, respectively. Converted in terms of molar units (of amino groups and pure dye molecules), it corresponds to molar ratio of 2, 2.7 and 3.1 mol amino groups per mole of dye. These values are signicantly lower (23 times) than those obtained at saturation of chitosan in the case of sorption experiments. Based on the number of sulphonyl groups of the dye, the ratio between amino groups and sulphonyl groups at the optimum dosage of coagulant varied between 0.5 and 0.75; this is much less than in the case of

Chitosan interactions with mercury and dyes the use of chitosan in the solid-state (i.e. 1.75). The other advantage of the coagulationocculation process vs. adsorption is due to the kinetics of the process. With the coagulation process the time required for the separation of the dye from the solution took about 3040 min, corresponding to pH control, fast agitation (for homogenizing the solution) and slow agitation (for maturation of the ocs) and nal ltration; this is much less than the time required for sorption process that required 2 h for smallest particles to more than 48 h for largest particles. The dissolving of the polymer results in the destruction of the crystallinity of the polymer, the breakage of hydrogen bonds between polymer chains makes amino groups more available and more accessible: this improves the saturation level of the polymer and allows a greater use of the number of amino groups present on the biopolymer. The well-opened structure of the polymer in the dissolved-state makes easier the accessibility to reactive sites and the kinetics of the process is greatly improved.

919
Chen, J.P. & Wang, L. 2001 Characterization of a ca-alginate based ion-exchange resin and its application in lead, copper, and zinc removal. Separation Science and Technology 36, 36173637. Chiou, M.-S. & Li, H.-Y. 2002 Equilibrium and modeling of adsorption of reactive dye on cross-linked chitosan beads. Journal of Hazardous Materials B93, 233248. Dambies, L., Vincent, T., Domard, A. & Guibal, E. 2001 Preparation of chitosan gel beads by ionotropic molybdate gelation. Biomacromolecules 2, 11981205. Dubois, M.A., Dozol, J.F. & Massiani, C. 1995 Pyrolysis and incineration of cationic and anionic ion-exchange resins identication of volatile degradation compounds. Journal of Analytical and Applied Pyrolysis 31, 129. Dzul Erosa, M.S., Saucedo Medina, T.I., Navarro Mendoza, R., Avila Rodriguez, M. & Guibal, E. 2001 Cadmium sorption on chitosan sorbents: kinetic and equilibrium studies. Hydrometallurgy 61, 157167. Gibbs, G., Tobin, J.M. & Guibal, E. 2003 Sorption of acid green 25 on chitosan: inuence of experimental parameters on uptake kinetics and sorption isotherms. Journal of Applied Polymer Science 90, 10731080. Gibbs, G., Tobin, J.M. & Guibal, E. 2004 Inuence of chitosan preprotonation on reactive black 5 sorption isotherms and kinetics. Industrial and Engineering Chemical Research 43, 111. Guibal, E. 2004 Metal ion interactions with chitosan a review. Separation and Purication Technology 38, 4374. Huang, C., Chen, S. & Pan, J.R. 2000 Optimal condition for modication of chitosan: a biopolymer for coagulation of colloidal particles. Water Research 34, 10571062. Huang, C. & Chen, Y. 1996 Coagulation of colloidal particles in water by chitosan. Journal of Chemical Technology and Biotechnology 66, 227232. Juang, R.-S. & Chiou, C.-H. 2000 Ultraltration rejection of dissolved ions using various weakly basic water-soluble polymers. Journal of Membrane Science 177, 207214. Juang, R.-S., Tseng, R.-L., Wu, F.-C. & Lee, S.-H. 1997 Adsorption behavior of reactive dyes from aqueous solutions on chitosan. Journal of Chemical Technology and Biotechnology 70, 391399. Kawamura, Y., Mitsushashi, M., Tanibe, H. & Yoshida, H. 1993 Adsorption of metal ions on polyaminated highly porous chitosan chelating resin. Industrial and Engineering Chemical Research 32, 386391. Krajewska, B. 2001 Diffusion of metal ions through gel chitosan membranes. Reactive and Functional Polymers 47, 3747. Kuncoro, E.K., Roussy, J. & Guibal, E. 2004 Mercury recovery by polymer-enhanced ultraltration: comparison of chitosan and poly(ethyleneimine) used as macro-ligand. Separation Science and Technology in press, McCarrick, P., Tobin, J. & Guibal, E. 2003 Comparative sorption of dyes on chitosan and activated carbon. Separation Science and Technology 38, 30493073. McKay, G., Blair, H.S. & Gardner, J.R. 1987 Two resistance mass transport model for the adsorption of acid dye onto chitin in xed beds. Journal of Applied Polymer Science 33, 12491257. Mi, F.-L., Shyu, S.-S., Lee, S.-T. & Wong, T.-B. 1999 Kinetic study of chitosan-tripolyphosphate complex reaction and acid-resistive properties of the chitosan-tripolyphosphate gel beads prepared by in-liquid curing method. Journal of Polymer Science Part B. Polymer Physics 37, 15511564. Pan, J.R., Huang, C., Chen, S. & Chung, Y.-C. 1999 Evaluation of a modied chitosan biopolymer for coagulation of colloidal particles. Colloids and Surfaces A: Physicochemical and Engineering Aspects 147, 359364. Piron, E., Accominotti, M. & Domard, A. 1997 Interaction between chitosan and uranyl ions. Role of physical and physicochemical parameters on the kinetics of sorption. Langmuir 13, 16531658. Roussy, J., Van Vooren, M. & Guibal, E. 2004 Chitosan for the coagulation and occulation of mineral colloids. Dispersion Science and Technology 25, 663677.

Conclusions Chitosan is very efcient for the binding of mercury and RB5 from dilute solutions. The poor diffusion properties of chitosan in the solid-state cause a control of uptake performance by sorbent particle size, especially in terms of kinetic behaviour. However, the solubility properties of chitosan in acidic solutions makes it possible to be used in the liquid form by coupling chelation mechanism to ultraltration for the recovery of mercury, or by using electrostatic attraction for the charge neutralization of sulphonyl groups (deprotonated at intermediary acid pH, i.e. pH 3) and the coagulationocculation of anionic dyes. Using chitosan in the liquid-form increases the accessibility to reactive sites (improved process kinetics) and/or their availability. The breaking of hydrogen bonds between amino groups and between hydroxyl groups (inter-chain or intra-chain bonds) during polymer dissolving much more available for interacting with metal ions and anionic dyes. This may explain the much more efcient use of amino groups of chitosan when the polymer is used in the dissolved-state for dye removal, as calculated by the molar ratio dye/amino groups at the saturation of the polymer. This effect is less signicant when considering mercury: in that case the positive effect of using chitosan in the dissolved-state (ultraltration coupled with chelation) is only signicant for binding kinetics since at saturation binding capacities tend to values of the same order of magnitude.

References
Annadurai, G., Juang, R.-S. & Lee, D.-J. 2002 Use of cellulose-based wastes for adsorption of dyes from aqueous solutions. Journal of Hazardous Materials B92, 263274.

920
Ruiz, M., Sastre, A.M. & Guibal, E. 2000 Palladium sorption on glutaraldehyde-crosslinked chitosan. Reactive and Functional Polymers 45, 155173. ` Sorlier, P., Denuziere, A., Viton, C. & Domard, A. 2001 Relation between the degree of acetylation and the electrostatic properties of chitin and chitosan. Biomacromolecules 2, 765772.

E. Guibal et al.
Wong, Y.C., Szeto, Y.S., Cheung, W.H. & McKay, G. 2004 Adsorption of acid dyes on chitosan-equilibrium isotherm analyses. Process Biochemistry 39, 695704. Yoshida, H. & Takemori, T. 1997 Adsorption of direct dye on crosslinked chitosan ber: breakthrough curve. Water Science and Technology 35, 2937.