Journal of Hazardous Materials 146 (2007) 65–72

Biosorption and bioreduction of Cr(VI) by a microalgal

isolate, Chlorella miniata
Xu Han a , Yuk Shan Wong b , Ming Hung Wong c , Nora Fung Yee Tam a,∗
a
Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong, China
b Department of Biology, Hong Kong University of Science and Technology, Kowloon, Hong Kong, China
c Croucher Institute of Environmental Sciences, Hong Kong Baptist University, Kowloon, Hong Kong, China

Received 25 May 2006; received in revised form 19 October 2006; accepted 26 November 2006
Available online 1 December 2006

Abstract
The ability and mechanism of a microalgal isolate, Chlorella miniata to remove Cr(VI) were investigated. Kinetic studies indicated that both
biosorption and bioreduction were involved in the Cr(VI) removal. The adsorbed Cr(VI) was reduced to Cr(III), and desorption studies indicated
that Cr(III) occupied most of the adsorption sites on the biomass. The equilibrium time for Cr(VI) removal was dependent on various factors
including initial pH, biomass and Cr(VI) concentrations. Equilibrium study showed that the Cr(VI) removal capacity was negatively related to the
initial pH, and the biosorption capacity of total Cr [Cr(III) and Cr(VI)] reached the maximum at initial pH of 3.0. The spectrum of Fourier Transform
Infrared Spectrometer analysis (FTIR) further confirmed that amino group on the algal biomass was the main adsorption site for Cr(VI) biosorption
in acidic pH while the reduced Cr(III) was mainly sequestered by carboxylate group. The comparison between biosorption–bioreduction and direct
bioreduction kinetic models proved that biosorption of Cr(VI) was the first step, followed by Cr(VI) bioreduction and Cr(III) biosorption on the
algal biomass.
© 2006 Elsevier B.V. All rights reserved.

Keywords: Algae; Biosorption; Hexavalent chromium; Trivalent chromium; Bioreduction

1. Introduction Cr(III), Cr(VI) as well as its other forms is regulated below

2 mg L−1 [3].
Chromium pollution in our environment has attracted more Many conventional methods such as chemical precipitation,
and more attention in recent years because of its harmful effects membrane separation, ion exchange and evaporation have been
to ecosystems and human beings. Hexavalent and trivalent are employed to remove Cr(VI) in industrial wastewater but they
two stable states of chromium in nature. These chromium species are not effective at metal concentrations ranging from 1 to
are commonly found in wastewater produced from leather 100 mg L−1 [4]. The high cost of the chemical reagents and the
tanning, dye, wood preservation and electroplating industries problems of secondary pollution also make the above physico-
and their concentrations could range from tens to hundreds chemical methods rather limited in application. In the last two
of mg L−1 [1]. Hexavalent chromium Cr(VI) is more toxic decades, more interests have been focused on using different
than the trivalent form Cr(III) because of its carcinogenic biosorbents to remove metal ions [5]. Among biosorbents, green
and mutagenic effects. A variety of diseases such as bron- algae are attractive as they are ubiquitous in natural environment,
chogenic carcinoma, asthma, pneumonitis and dermatitis have have large surface area to volume ratio and high binding affinity
been reported to associate with occupational Cr(VI) exposure to pollutants [6]. Chlorella miniata, a green microalgal species,
[2]. Hence, the discharge of Cr(VI) to surface water is regulated with a spherical to ellipsoidal shape (diameter around 2–3 ␮m
below 0.05 mg L−1 by the U.S. EPA, and total Cr including with a surface area to volume ratio of 1.1) was isolated from
a municipal sewage treatment plant in Hong Kong SAR by the
present research team. Our studies showed that this isolate had
∗ Corresponding author. Tel.: +852 27887793; fax: +852 27887406. a high growth rate in domestic wastewater, and its high biosorp-
E-mail address: [email protected] (N.F.Y. Tam). tion capacity to Ni(II) and Zn(II) ions from contaminated water

0304-3894/$ – see front matter © 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.jhazmat.2006.11.053
66 X. Han et al. / Journal of Hazardous Materials 146 (2007) 65–72

had been reported [6–8]. However, its ability in removing Cr(VI) with deionized water twice to remove any residues adsorbed on
has never been studied. the cell surfaces. The washed cells were then freeze-dried and
The mechanism involved in the removal of Cr(VI) is com- grounded into fine particles prior to biosorption experiments.
plex and depends on the properties of biosorbents. Previous
studies claimed that the removal of Cr(VI) by biomass was 2.2. Preparation of Cr(VI) solution
mainly through ion exchange and binding on functional groups
[9]. However, the appearance of Cr(III) in solution suggested The stock solution of Cr(VI) (1000 mg L−1 ) was prepared
that Cr(VI) adsorption along with its reduction to Cr(III) may in deionized water with potassium dichromate (K2 Cr2 O7 ). All
have occurred during the uptake process [3,10–14]. Different working concentrations were obtained by diluting the stock solu-
mechanisms including ion exchange-redox reaction [12], par- tion with deionized water, and pH was adjusted to the desired
allel biosorption and bioreduction [13], direct reduction and a values according to the following experimental design with 1 M
sequential three-step [14] were proposed. The mechanism of HCl and 1 M NaOH solutions.
Cr(VI) removal by the microalgal species isolated from wastew-
ater may be different from other biosorbents due to the difference 2.3. Kinetic experiments
in biomass composition.
Various kinetic models for Cr(VI) removal have been The equilibrium time of Cr(VI) removal was determined
proposed, however, they are not correlated well with the cor- under different initial pH (from 1.0 to 4.0) and biomass concen-
responding mechanisms. The pseudo-first order kinetic model, trations (from 1.0 to 5.0 g L−1 ) at an initial Cr(VI) concentration
assuming only adsorption took place and without any bioreduc- of 100 mg L−1 . The effect of initial Cr(VI) concentrations on the
tion, has been widely used in Cr(VI) removal [15]. Park et al. kinetic process was investigated by another experiment using
proposed a second order kinetic model based on Cr(VI) reduc- 5.0 g L−1 biomass, initial pH 2.0, and varied initial Cr(VI) con-
tion but their model showed little correlation with the proposed centrations, 20, 60 and 100 mg L−1 . In all experiments, the
mechanism [14,16]. Although a parallel reduction and adsorp- working volume was 150 mL in a 250 mL conical flask agitated
tion kinetic model has been proposed for Cr(VI) removal by on a shaker at 160 rpm at room temperature (25 ± 1 ◦ C). Liquid
Cabatingan et al. [13], their results showed that increasing the solution samples (2 mL from each flask) were collected at reg-
rate of adsorption would lead to increase of reduction and vice ular time intervals and analyzed for residual concentrations of
versa, which is the character of consecutive reaction rather than Cr(VI) and total chromium.
parallel reaction, indicating that something must be wrong in
their model. It is necessary to develop a new kinetic model based
on the Cr(VI) removal mechanism. The present study therefore 2.4. Equilibrium experiments
aims to: (i) evaluate the mechanism involved in the removal of
Cr(VI) by a local microalgal isolate, C. miniata; (ii) understand Equilibrium experiments were carried out to investigate
the quantitative relationship between biosorption and bioreduc- the effect of initial pH on the Cr(VI) removal process. Algal
tion in Cr(VI) removal through a series of kinetic, equilibrium biomass 2.0 g L−1 was mixed with water containing 50, 100 and
and desorption studies; (iii) develop kinetic models based on the 200 mg L−1 Cr(VI) at initial pH varied from around 0 to 4.0.
biosorption–bioreduction mechanism; (iv) identify the possible The flasks were agitated on a shaker at 160 rpm for 12 days to
sorption sites that were involved in the Cr(VI) removal pro- ensure that the reaction would reach equilibrium. Samples were
cess using the Fourier Transform Infrared Spectrometer analysis then centrifuged at 3500 rpm for 10 min, and the supernatant
(FTIR). was used for determination of Cr(VI) and Cr(III).

2. Materials and methods 2.5. Desorption experiments

2.1. Mass culture of microalgae and preparation of The Cr-loaded biomass obtained from the above equilibrium
biosorbent experiments was treated with three different desorbents, namely
deionized water, 0.5 M HCl and 0.5 M NaOH to elute Cr from
C. miniata was cultivated in a transparent acrylic column the biomass. The working volume was 20 mL and the flasks were
(internal diameter of 140 mm and length of 100 cm) containing agitated on a shaker at 160 rpm for 24 h. After desorption, sam-
approximate 10 L Bristol medium. The composition of the Bris- ples were centrifuged at 3500 rpm for 10 min and the supernatant
tol medium was (g L−1 medium): NaNO3 , 25; K2 HPO4 , 7.5; was analyzed for Cr(VI) and total Cr concentrations.
KH2 PO4 , 17.5; MgSO4 ·7H2 O, 11.8; NaCl, 2.5; CaCl2 ·2H2 O,
2.5; FeCl3 ·6H2 O, 0.5; MnCl2 ·4H2 O, 0.03; CoCl2 ·6H2 O, 0.002; 2.6. FTIR analysis
CuSO4 ·5H2 O, 0.001; ZnSO4 ·7H2 O, 0.004; NaMoO4 ·2H2 O,
0.002 and EDTA, 0.54 (acid form). The culture was illuminated Infrared spectra of the control (biomass without Cr(VI) treat-
by cool fluorescent light with an average light intensity 4.2 klux ment) and the biomass mixed with 400 mg L−1 Cr(VI) at initial
in 16-h light:8-h dark cycle at room temperature 25 ± 1 ◦ C. After pH 2.0 for 12 days were obtained using a Fourier Transform
reaching the stationary phase, the cells were harvested and cen- Infrared Spectrometer (Nocolet, Avatar E.S.P.360). A mea-
trifuged at 5000 rpm for 15 min, the cell pellets were washed sured amount of biomass was mixed with KBr (2% potassium
 X. Han et al. / Journal of Hazardous Materials 146 (2007) 65–72 67

bromide). The mixture was grounded into fine particles and com- both initial pH and biomass concentrations (Fig. 1). A rapid
pressed into translucent sample disks by a manual hydraulic removal of Cr(VI) took place in the first 30 min, and the rate
press. The disks were then fixed in the FTIR spectrometer for became level off thereafter. Low initial pH as well as high
analysis. biomass shortened the equilibrium time and enhanced the Cr(VI)
removal percentages. At initial pH 1.0, biomass 2.0 g L−1 , nearly
2.7. Analysis of chromium 100% Cr(VI) was removed within 58 h. At initial pH 4.0, less
than 10% Cr(VI) was removed and it was impossible to estimate
Cr(VI) and total Cr in liquid solution were determined accord- the equilibrium time (Fig. 1a). The Cr(VI) removal percentages
ing to the standard method described by Clesceri et al. [17] and at initial pH 2.0 were 60, 85 and 100% in treatments with 1.0,
Kratochvil et al. [12]. The absorbance of the purple complex 2.0 and 5.0 g L−1 biomass, respectively, and the respective equi-
formed from reacting Cr(VI) with 1,5-diphenylcarbohydrazide librium time were 240, 215 and 150 h (Fig. 1b). Previous results
was measured at λ = 540 nm by a UV spectrophotometer (Shi- also reported that the equilibrium time of Cr(VI) removal by sea-
madzu, UV-1201) and the detection limit was 0.05 mg L−1 . weed and fungi varied from tens to hundreds of hours depending
Total chromium including Cr(VI) and Cr(III) was determined by on experimental conditions [3,14,16].
atomic absorption spectroscopy (AAS) (Shimadzu, AA-6501) Equilibrium time was also dependent on initial Cr(VI) con-
at λ = 357.9 nm and the detection limit of AAS in the present centrations. The respective equilibrium time under initial Cr(VI)
study was 0.1 mg L−1 . The Cr(III) content in liquid solution concentrations of 100, 60 and 20 mg L−1 was 150, 72 and 30 h,
was obtained by subtracting the content of Cr(VI) from that of respectively (Fig. 2). Cr(III) appeared gradually with the removal
total chromium. of Cr(VI), indicating that the Cr(VI) adsorbed on the algal
biomass was reduced to Cr(III). The amounts of Cr(VI) removed
3. Results and discussion from the contaminated water were more than the amounts of
Cr(III) detected, suggesting that not all of the biosorbed Cr(VI)
3.1. Kinetic studies of Cr(VI) removal was reduced to Cr(III), some of the reduced Cr(III) was released
to the liquid solution while some adsorbed on the biomass. The
Kinetic results showed that the removal of Cr(VI) by C. mini- biosorption and bioreduction processes were likely to be phys-
ata and the equilibrium time were significantly dependent on icochemical transformation as the biomass used in the present

Fig. 1. Kinetic study of Cr(VI) removal under (a) different initial pH and (b) biomass dosage (initial Cr(VI) concentration 100 mg L−1 , volume 150 mL; mean and
standard deviation values of three replicates are shown).
68 X. Han et al. / Journal of Hazardous Materials 146 (2007) 65–72

Cr(VI) forms HCrO4 − , Cr2 O7 2− , CrO4 2− , HCr2 O7 − and

H2 CrO4 in solution, and the relative proportion of each species
depends on both pH and Cr(VI) concentration [2]. Cabatingan et
al. [13] suggested in the pH range of 1–6.5 and a total Cr(VI) con-
centration of 7.69 × 10−3 M, HCrO4 − was the most dominant
form with coexistence of H2 CrO4 , Cr2 O7 2− and HCr2 O7 − in
solution, but Cr(VI) was mainly present as the CrO4 2− anion
when pH was larger than 6.5. Nieboer and Jusys [18] also
found that HCrO4 − was the predominant form up to the Cr(VI)
concentration of 10−2 M, and it started to condense yielding
the orange-red dichromate ion (Cr2 O7 2− ) above this concen-
tration. In the present study, the highest Cr(VI) concentration
was 200 mg L−1 (3.85 × 10−3 M), therefore, in the pH range
of 1.0–4.0, HCrO4 − should be the dominant Cr(VI) species in
solution. In neutral or alkali solution, it is difficult for the anion
species of Cr(VI) binding to the negatively charged functional
groups on the biomass surface. However, in acidic pH, the func-
tional groups on the biomass were protonated and positively
charged, thus became available for Cr(VI) anion biosorption.
Lowering pH resulted in higher Cr(VI) biosorption due to the
increase of electrostatic attraction between sorbate and the proto-
nated groups on the biomass. Similar trend has also been found in
Cr(VI) removal by fungal biomass [14], crab shell [19] and chi-
tosan [20], and in biosorption of other anions such as Au(CN)2 − ,
SeO4 2− and VO4 2− [19].
At low pH, on the other hand, Cr(VI) had a high redox poten-
tial and favored Cr(VI) bioreduction [12]. In addition, reductants
on the biomass such as carbohydrate and protein could supply
electrons for Cr(VI) bioreduction, with partial release of soluble
organics or ultimate oxidized product, CO2 [3]. This explains
why increasing biomass dosage and lowering pH of the contam-
inated water could achieve more Cr(VI) removal within a shorter
period of time.

3.2. Effect of initial pH on Cr(VI) removal at equilibrium

Equilibrium experiments were conducted at an initial pH

varied from around 0 to 4.0 under initial Cr(VI) concentrations
of 50, 100 and 200 mg L−1 for 12 days. The effects of initial pH
on Cr(VI) removal and total Cr bisorption capacity were similar
at different initial Cr(VI) concentrations. At equilibrium, the
pH changed from initial values of 1.0, 2.0 and 4.0 to 1.03, 2.25
Fig. 2. Cr species and distribution in solution during the removal of Cr(VI) from
and 5.79, respectively (data not shown). Fig. 3a shows that
contaminated water under three initial Cr(VI) concentrations: (a) 100 mg L−1 ,
(b) 60 mg L−1 and (c) 20 mg L−1 (biomass dosage 5.0 g L−1 , initial pH 2.0, the Cr(VI) removal capacity of C. miniata decreased linearly
volume 150 mL; average of duplicates are shown). with increases of initial pH. At initial pH 3.0, 35.42, 38.02 and
40.72% of Cr(VI) at concentrations of 200, 100 and 50 mg L−1
were removed, respectively. When initial pH approached 0
study was freeze-dried, grounded into fine particles, and might and H+ concentration reached 1.0 M, Cr(VI) was completely
have lost its biological activity. The pattern was similar among removed. This could be attributed to the easier biosorption
different initial Cr(VI) concentrations. Zhao and Duncan [11] of Cr(VI) on the protonated biosorbents [20] and the higher
also reported that Cr(VI) removal was probably a bioreduction reduction–oxidation potential of Cr(VI) at lower pH as reported
along with a biosorption process. Fig. 2 further revealed that the by Kratochvil et al. [12].
concentration of total Cr [Cr(VI) and Cr(III)] dropped signifi- The bisorption capacity of total Cr [Cr(VI) and Cr(III)] of
cantly in the first few hours and reached equilibrium at 4–6 h, C. miniata was also a pH dependent process, and the maxi-
and the equilibrium time was much shorter than that of Cr(VI) mum capacity was obtained at an initial pH 3.0 under all three
or Cr(III), suggesting that Cr(VI) bioreduction process was the initial Cr(VI) concentrations (Fig. 3b). Although low pH (0–1)
rate limiting step in the removal of Cr(VI) by C. miniata. could increase Cr(VI) biosorption on the protonated biomass
 X. Han et al. / Journal of Hazardous Materials 146 (2007) 65–72 69

Fig. 3. Initial pH effect on (a) Cr(VI) removal capacity and (b) total Cr biosorp- Fig. 4. Effects of different desorbents on (a) Cr(VI) and (b) total Cr recov-
tion capacity under different initial Cr(VI) concentrations (biomass dosage ery under different initial Cr(VI) concentrations (biomass dosage 5.0 g L−1 ,
2.0 g L−1 , volume 20 mL; mean and standard deviation values of three replicates desorbent volume 20 mL).
are shown).
around 0.3) could be attributed to the further reduction of the
and bioreduction of Cr(VI) to Cr(III), the reduced Cr(III) was adsorbed Cr(VI) to Cr(III) under acidic condition.
difficult to be adsorbed on the biomass due to electric repul- Alkali wash by 0.5 M NaOH also had significantly higher
sion leading to low total Cr biosorption. On the other hand, at percentages of total Cr recovery than the other two biosorbents
high pH such as 3.0 or larger, less Cr(III) was produced due to (Fig. 4b). When NaOH was used as the desorbent, 70–100% of
the sharp decrease of both Cr(VI) biosorption and bioreduction total Cr were recovered while the pecentages of Cr(VI) desorbed
processes. As a consequence, most Cr(VI) would still remain in were very low (less than 15%), suggesting that most Cr adsorbed
the contaminated water and the biosorption of total Cr was also on the biomass was in the form of Cr(III) and Cr(OH)4 − was
low. Previous studies reported that the optimal pH for total Cr probably the main composition in NaOH elutant based on the
biosorption was around 2–3. For instance, the optimal pH for amphotericity of Cr(III). The recovery performance of 0.5 M
Sphagnum-moss peat, leaf, mould and coconut-husk fibre were HCl was not better than deionized water, indicating protons
1.5, 2.0, 2.05 and 2.0, respectively [11], while 2–2.5 was the opti- could not replace Cr(III) adsorbed on the biomass.
mal pH for Sargassum [12,13]. Park et al. [3] further showed that
the optimal pH for total Cr biosorption would change according 3.4. FTIR analysis
to contact time, the optimal pH was 1.5–2.5 after 6 h contact and
changed to pH 4.0 after 480 h (the time for a complete reaction) The infrared spectra of the control biomass of C. miniata
in Ecklonia sp., probably due to the release of organic matter (not subject to any chromium treatment) and the biomass after
which complexed with Cr(III). mixing with 400 mg L−1 Cr(VI) at initial pH 2.0 were shown
in Fig. 5. The absorption peaks at 1654 and 1540 cm−1 cor-
3.3. Desorption studies of Cr species adsorbed on biomass responded to the amide I and amide II bands, respectively, as
suggested by Yee et al. [21] were found in both control and
When the three desorbents were used to elute Cr from the treated algal biomass in the present study. Glucosamine group
algal biomass, more Cr(VI) were desorbed by 0.5 M NaOH than had been reported as an important sugar component of the rigid
that by deionized water and 0.5 M HCl (Fig. 4a). Since Cr(VI) wall in many Chlorella species [22]. Under low pH, amino group
adsorbed on the biomass was due to proton bridge, when pro- could be protonated and thus responsible for Cr(VI) adsorption
tons were consumed in alkali wash, the adsorbed Cr(VI) would [20,23]. The region between 3200 and 3500 cm−1 represented
be released. Boddu et al. [20] also found that NaOH was an the overlapping peaks of stretching vibration of O–H and N–H
effective desorbent to elute Cr(VI) adsorbed on chitosan. The [24]. The region between 3000 and 2800 cm−1 exhibited the
least Cr(VI) recovery percentage by acid wash (0.5 M HCl, pH C–H stretching vibrations of –CH3 and CH2 functional groups,
70 X. Han et al. / Journal of Hazardous Materials 146 (2007) 65–72

or other reducing organic matters; (iii) release of Cr(III): part

of the bioreduced Cr(III) was released from the biomass.
To further confirm the above mechanism, two kinetic models,
the biosorption–bioreduction model and the direct bioreduction
model were developed and compared in the present study. The
chemical equation of the first model could be defined:
HCrO4 − + H+ + Biomass ⇔ HCrO4 − − H+ − Biomass
→ Cr3+ + H2 O + Biomass (oxidized) (1)
Since HCrO4 − biosorption on biomass was a fast step com-
pared to bioreduction, the reaction rate was determined by the
bioreduction step. If bioreduction of Cr(VI) on the biomass was
thought to be a pseudo-first order reaction, it could be defined:
Fig. 5. FTIR analysis of Cr(VI)-treated and control biomass (labels 1, 2, 3 and
4 represent the changes on the algal biomass after Cr(VI) treatment). dq
= −k1 q (2)
dt
and 1300–1470 cm−1 was the deformation stretching of C–H, where k1 is the apparent reaction rate constant for the
–CH3 , and CH2 functional groups [21]. At 1400 cm−1 , it was biosorption–bioreduction model and q is the Cr(VI) adsorbed
a characteristic peak of symmetric vibrational COO− frequen- on biomass.
cies of terminal amino acid on biomass [25]. The peaks of 1240, If Cr(VI) sorption equilibrium was thought to be present
1076 cm−1 represented P O and C–O bands of polysaccharides, during the whole process, q could be expressed by Langmuir
respectively [26]. isotherm:
After Cr(VI) treatment, four changes of the functional groups QbC
on the biomass were detected from the spectrum. The first q= (3)
bC + 1
change was the enhancement of the intensity at the region
3200–3500 cm−1 , indicating an increase of the free hydroxyl where Q is the maximum sorption capacity, b the sorption con-
group on the biomass (Fig. 5). This could be due to hydrolyzing stant of Cr(VI) and C is the concentration of Cr(VI) in solution.
of some polysaccharides on the cell wall to shorter saccharides By combining Eqs. (2) and (3), the following equation was
such as oligosaccharides, dioses, and monoses under acidic con- obtained:
dition [25]. The second change was the weakening of the peak dC
at 1400 cm−1 , which was typical of the complexation of the car- = −k1 C(bC + 1) (4)
dt
boxylate functional group by coordination with metal cations
[25]. In the present study, this might be due to the complexation with the initial conditions: t = 0, C = C0 , C0 is the initial concen-
of Cr(III) formed from bioreduction of Cr(VI) with carboxylate tration of Cr(VI). After a definite integral, we could get:
group. Our previous study confirmed that Cr(III) was mainly    
C0 C
sequestered by carboxylate group on C. miniata [27]. The third k1 t = ln − ln (5)
bC0 + 1 bC + 1
change was the shift of the peak at 1076–1044 cm−1 , which
could be due to the involvement of the C–O bond of polysaccha- In the direct bioreduction kinetic model, a pseudo-first order
rides in Cr(III) biosorption. Similar study on using grape stalks reaction was assumed due to its simple form as described by
to adsorb copper and nickel ions also suggested that lignin C–O Sparks [30], and the following equation could be defined:
might be involved in metal uptake [28]. The last change was the
dC
presence of a new peak at around 940 cm−1 in the Cr(VI) treated = −k2 C (6)
biomass, and it could be attributed to the presence of Cr(VI)–O dt
bond as suggested by Holman et al. [29]. where k2 is the apparent reaction rate constant for the direct
bioreduction. Then we could get:
3.5. Mechanism and kinetic modeling of Cr(VI) biosorption k2 t = ln(C0 ) − ln(C) (7)
and bioreduction
The parameters k1 , b in Eq. (5) and k2 in Eq. (7) were
Based on the above results, it is reasonable to conclude that estimated by a nonlinear regression using the Sigmaplot 8.0
the mechanism involved in the removal of Cr(VI) by C. miniata software and the results are listed in Table 1. The rate con-
was biosorption–bioreduction. The sequence was: (i) biosorp- stant k2 in the direct bioreduction model was larger than k1 in
tion of Cr(VI) at low pH: protons were adsorbed on the amino the biosorption–bioreduction model, which could be attributed
group of the algal biomass, the Cr(VI) ions were then adsorbed to the combined effect of biosorption and bioreduction in
on the protonated sites; (ii) bioreduction of Cr(VI) to Cr(III): direct bioreduction model. The higher R2 of the biosorption–
the Cr(VI) adsorbed on the biomass surface was bioreduced to bioreduction model than the direct bioreduction one indicated
Cr(III) by the reductants on the biomass such as polysaccharides the former model was more likely to be involved in the removal
 X. Han et al. / Journal of Hazardous Materials 146 (2007) 65–72 71

Table 1
Regression parameters of Cr(VI) removal under different Cr(VI) initial concentration by Chlorella miniata
Cr(VI) initial Parameters in biosorption–bioreduction model Parameters in direct bioreduction model
concentration (mg L−1 )
k1 × 102 (h−1 ) b × 102 (L mg−1 ) R2 k2 × 102 (h−1 ) R2

100 2.15 6.99 0.973 4.78 0.836

60 5.58 9.15 0.889 7.54 0.847
20 15.24 20.25 0.982 26.27 0.872

of Cr(VI) by C. miniata. It further confirmed that Cr(VI) biosorp- [6] A.M.Y. Chong, Y.S. Wong, N.F.Y. Tam, Performance of different microal-
tion on the biomass was the first step and it was then bioreduced gal species in removing nickel and zinc from industrial wastewater,
to Cr(III). The increase of k1 and b values with decreases of Chemosphere 41 (2000) 251–257.
[7] J.P.K. Wong, Y.S. Wong, N.F.Y. Tam, Nickel biosorption by two Chlorella
initial Cr(VI) concentrations (Table 1) could be explained by species, C. vulgaris (a commercial species) and C. miniata (a local isolate),
the fact that less proton was consumed in water with lower Bioresour. Technol. 73 (2000) 133–137.
Cr(VI) contamination and the pH was maintained at lower level [8] N.F.Y. Tam, J.P.K. Wong, Y.S. Wong, Repeated use of two Chlorella
throughout the study, thus was easier for Cr(VI) biosorption and species, C. vulgaris and WW1 for cyclic nickel biosorption, Environ. Pollut.
bioreduction. 114 (2001) 85–92.
[9] G. Donmez, Z. Aksu, Removal of chromium(VI) from saline wastewaters
by Dunaliella species, Process Biochem. 38 (2002) 751–762.
4. Conclusions [10] D.C. Sharma, C.F. Forster, Removal of hexavalent chromium using Sphag-
num moss peat, Water Res. 27 (1993) 1201–1208.
The present study shows that the green microalgal isolate, [11] M. Zhao, J.R. Duncan, Batch removal of sexivalent chromium by Azolla
filiculoides, Biotechnol. Appl. Biochem. 26 (1997) 179–182.
C. miniata was capable of removing Cr(VI) from the contam-
[12] D. Kratochvil, P. Pimentel, B. Volesky, Removal of trivalent and hexava-
inated water. At an initial pH of 2.0 and biomass of 5.0 g L−1 , lent chromium by seaweed biosorbent, Environ. Sci. Technol. 32 (1998)
65% Cr(VI) was removed from the contaminated water con- 2693–2698.
taining 100 mg Cr(VI) L−1 in the first 2 h, while a complete [13] L.K. Cabatingan, R.C. Agapay, J.L.L. Rakels, M. Ottens, L.A.M. van der
Cr(VI) removal was obtained at 150 h. The main adsorption Wielen, Potential of biosorption for the recovery of chromate in industrial
wastewaters, Ind. Eng. Chem. Res. 40 (2001) 2302–2309.
site for Cr(VI) in acidic pH was amino group, and the reduced
[14] D. Park, Y.S. Yun, J.H. Jo, J.M. Park, Mechanism of hexavalent chromium
Cr(III) was mainly sequestered by carboxylate group on the removal by dead fungal biomass of Aspergillus niger, Water Res. 39 (2005)
algal biomass. The kinetic model developed based on the 533–540.
biosorption–bioreduction mechanism had a significantly higher [15] D. Mohan, K.P. Singh, V.K. Singh, Removal of hexavalent chromium from
R2 than that of the direct bioreduction model, further confirmed aqueous solution using low-cost activated carbons derived from agricultural
waste materials and activated carbon fabric cloth, Ind. Eng. Chem. Res. 44
that the removal of Cr(VI) was based on Cr(VI) biosorption,
(2005) 1027–1042.
bioreduction of Cr(VI) to Cr(III) and Cr(III) biosorption on the [16] D. Park, Y.S. Yun, J.M. Park, Use of dead fungal biomass for the detoxifi-
biomass. cation of hexavalent chromium: screening and kinetics, Process Biochem.
40 (2005) 2559–2565.
Acknowledgements [17] L.S. Clesceri, A.E. Greenberg, A.D. Eaton, Standard Methods for the
Examination of Water and Wastewater, 20th ed., American Public Health
Association, Washington, DC, 1998, pp. 366–368.
The research work was supported by the Areas of Excellence [18] E. Nieboer, A.A. Jusys, Biologic chemistry of chromium, in: J.O. Nriagu,
Scheme established under the University Grants Committee of E. Nieboer (Eds.), Chromium in Natural and Human Environments, Wiley
the HKSAR (Project No. AoE/P-04/2004). The funding support Interscience, New York, 1988, pp. 21–81.
of the Research Centre for Coastal Pollution and Conservation, [19] H. Niu, B. Volesky, Characteristics of anionic metal species biosorption
with waste crab shells, Hydrometallurgy 71 (2003) 209–215.
City University of Hong Kong, was also acknowledged.
[20] V.M. Boddu, K. Abburi, J.L. Talbott, E.D. Smith, Removal of hexavalent
chromium from wastewater using a new composite chitosan biosorbent,
References Environ. Sci. Technol. 37 (2003) 4449–4456.
[21] N. Yee, L.G. Benning, V.R. Phoenix, F.G. Ferris, Characterization of
[1] E. Dermou, A. Velissariou, D. Xenos, D. Vayenas, Biological chro- metal-cyanobacteria sorption reactions: a combined macroscopic and
mium(VI) reduction using a trickling filter, J. Hazard. Mater. 126 (2005) infrared spectroscopic investigation, Environ. Sci. Technol. 38 (2004) 775–
78–85. 782.
[2] J. Kotas, Z. Stasicka, Chromium occurrence in the environment and meth- [22] H. Takeda, Diversity of cell-wall chemical composition and the taxonomy
ods of its speciation, Environ. Pollut. 107 (2000) 263–283. of algae, in: B.R. Chaudhary, S.B. Agrawal (Eds.), Cytology, Genetics
[3] D. Park, Y.S. Yun, J.M. Park, Reduction of hexavalent chromium with and Molecular Biology of Algae, SPB Academic, Amsterdam, 1996, pp.
the brown seaweed Ecklonia biomass, Environ. Sci. Technol. 38 (2004) 291–301.
4860–4864. [23] R.S. Bai, T.E. Abraham, Studies on enhancement of Cr(VI) biosorption by
[4] A. Kapoor, T. Viraraghavan, Fungal biosorption—an alternative treatment chemically modified biomass of Rhizopus nigricans, Water Res. 36 (2002)
option for heavy metal bearing wastewaters: a review, Bioresour. Technol. 1224–1236.
53 (1995) 195–206. [24] S. Deng, Y.-P. Ting, Characterization of PEI-modified biomass and
[5] T.A. Davis, B. Volesky, A. Mucci, A review of the biochemistry of heavy biosorption of Cu(II), Pb(II) and Ni(II), Water Res. 39 (2005) 2167–
metal biosorption by brown algae, Water Res. 37 (2003) 4311–4330. 2177.
72 X. Han et al. / Journal of Hazardous Materials 146 (2007) 65–72

[25] Z.Y. Lin, J.M. Wu, R. Xue, Y. Yang, Spectroscopic characterization of Au3+ [28] I. Villaescusa, N. Fiol, M. Martinez, N. Miralles, J. Poch, J. Serarols,
biosorption by waste biomass of Saccharomyces cerevisiae, Spectrochim. Removal of copper and nickel ions from aqueous solutions by grape stalks
Acta, Part A 61 (2005) 761–765. wastes, Water Res. 38 (2004) 992–1002.
[26] G. Benning, V.R. Phoenix, N. Yee, M.J. Tobin, Molecular charac- [29] H.Y.N. Holman, D.L. Perry, M.C. Martin, G.M. Lamble, W.R. McKinney,
terization of cyanobacterial silicification using synchrotron infrared J.C. Hunter-Cevera, Real-time characterization of biogeochemical reduc-
micro-spectroscopy, Geochim. Cosmochim. Acta 68 (2004) 729–741. tion of Cr(VI) on basalt surfaces by SR-FTIR imaging, Geomicrobiol. J.
[27] X. Han, Y.S. Wong, N.F.Y. Tam, Surface complexation mechanism and 16 (1999) 307–324.
modeling in Cr(III) biosorption by a microalgal isolate, Chlorella miniata, [30] D.L. Sparks, Kinetics of Soil Chemical Process, Academic Press, New
J. Colloid Interface Sci. 303 (2006) 365–371. York, 1989, 210 pp.