Bioresource Technology 102 (2011) 80358039

Contents lists available at ScienceDirect

Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

The use of a white rot fungi (Pleurotus ostreatus) immobilized on Amberlite XAD-4
as a new biosorbent in trace metal determination
Sevgi Kocaoba a,, Munevver Arsoy b
a
Yildiz Technical University, Faculty of Art and Science, Department of Chemistry, Davutpasa Cad., No: 127, 34210-Davutpasa, Istanbul, Turkey
b
Ankara University, Faculty of Health Sciences, Department of Nutrition and Dietary, Altndag, Ankara, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: The present work proposes the use of Pleurotus ostreatus immobilized on Amberlite XAD-4 as new biosor-
Received 8 February 2011 bent in trace metal determination. The effects of experimental parameters, such as pH and ow rate of
Received in revised form 2 May 2011 sample solution, amount of solid phase, eluent type, and concentration on the recovery of the metal ions
Accepted 3 May 2011
were investigated. Maximum adsorption of Cr(III), Cd(II) and Cu(II) ions took place in the pH range 45.
Available online 7 May 2011
These metal ions can be desorbed with 1 M HCl (recovery 95100%). 0.2 g adsorbent amount and
2.5 mL min 1 ow rate was found to be optimum of all preconcentration experiments. The sorption
Keywords:
capacity after 10 cycles of sorption and desorption does not vary more than 2.0%. The inuences of the
Chromium(III)
Cadmium(II)
contaminant ions on the retentions of the analytes were also examined. The results showed that P. ostre-
Copper(II) atus immobilized on Amberlite XAD-4 can be considered as very promising material in trace metal
Preconcentration determination.
Pleurotus ostreatus Crown Copyright 2011 Published by Elsevier Ltd. All rights reserved.

1. Introduction
Toxic heavy metal contamination of industrial wastewater is an
important environmental problem. Many industries, such as auto-
motive, metal nishing, electroplating, battery manufacturing,
mining, electric cable manufacturing, tannery, steel and textile
industries, and release various concentrations of heavy metals like
lead, copper, chromium, zinc, cadmium, and nickel, in wastewa-
ters. In addition to their toxicity effects even at low concentrations,
heavy metals can accumulate throughout the food chain, which
leads to serious ecological and health hazards as a result of their
solubility and mobility (Kocaoba, 2007).
Chromium is released into the environment by a large number
of industries such as mining, iron sheet cleaning, chrome plating,
leather tanning and wood preservation (Krishna and Philip,
2005). According to US EPA, the discharge of Cr(VI) into surface
water is 0.5 mg L 1, while total Cr including Cr(III), Cr(VI) and its
other forms is regulated to below 2 mg L 1 (Baral and Engelken,
2002).
The major sources of cadmium release into the environment by
waste streams are electroplating, smelting, alloy manufacturing,
pigments, plastic, battery, mining and rening processes (Tsezos,
2001). The permissible limits of cadmium discharge in wastewater
and drinking water are 0.1 and 0.05 mg L 1, respectively.
Corresponding author. Tel.: +90 212 383 4152.
E-mail addresses: kocaoba@yildiz.edu.tr, kocaobasevgi@hotmail.com (S. Kocao-
ba).
Since copper is a widely used material, there are many actual or
potential sources of copper pollution. Copper may be found as a
contaminant in food, especially shellsh, liver, mushrooms, nuts,
and chocolate. Briey, any processing or container involving cop-
per material may contaminate the products such as food, water
or drink (Prasad and Freitas, 2000).
The permissible limits in Turkeys wastewaters are 0.5, 0.05 and
0.5 mg L 1, for drinking water are 0.05, 0.005 and 0.02 mg L 1
respectively, chromium, cadmium and copper (Yaramaz, 1992).
In order to minimize the adverse effects of these heavy metals,
authorities and environmental agencies all over the world enforced
stringent standards for the maximum allowable limits of heavy
metals discharge into open landscapes and water bodies. These
strict regulations and standards encouraged researchers to search
for new technologies, which are environmentally friendly and
can reduce heavy metals concentrations in the discharged waste-
waters to be within the maximum allowable limits. The conven-
tional methods used to treat wastewaters containing heavy
metals include chemical precipitation, coagulation, ion exchange,
solvent extraction, membrane processes, reverse osmosis, and
adsorption. However, most of these methods have several disad-
vantages, such as: the high cost of treatment, the need for contin-
uous input of chemicals, the production of toxic sludge, or the
concentration of heavy metals in the treated wastewater may not
be below the maximum allowable limit (Abu Al-Rub et al., 2004).
Biosorption, which involves the use of biomass or natural sub-
stances as sorbents, presents an attractive alternative to the tradi-
tional physicochemical means for removing toxic heavy metal

0960-8524/$ - see front matter Crown Copyright 2011 Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2011.05.004
8036 S. Kocaoba, M. Arsoy / Bioresource Technology 102 (2011) 80358039
from ground and wastewaters (Saeed et al., 2005; Gong et al.,
2005; Davis et al., 2003; Holan and Volesky, 1995; Xu and Liu,
2008; Pavasant et al., 2006).
Biosorbents may be viewed as natural ion-exchange materials
that primarily contain weakly acidic and basic groups. The metals
can be stripped from the biomaterial after loading by sulfuric or
hydrochloric acid, sodium hydroxide or complexing agents,
whether dead biomass or live bacteria are used. The sensitivity of
living cells to extremes of pH or high metal concentration and need
to furnish metabolic energy are some of the major constraints of
employing growing cells for bioremediation. The major advantages
of the biosorption technology are its effectiveness in reducing the
concentration of heavy metal ions to very low levels and the use
of inexpensive biosorbent materials such as naturally abundant al-
gae or by products of fermentation industries (Holan et al., 1993).
There are evidences that some phenomena for metal binding, such
as adsorption, ion exchange, chelating, precipitation and crystalli-
zation take place in cellular membrane of microorganisms (Pardo
et al., 2003). The external factors such as pH, temperature, con-
centration of metal ion, and contact time always affect the sorp-
tion of metal ions. Fungi is better suitable for the removal of
metals from wastewater than other microbes because of their great
tolerance towards heavy metals and other adverse conditions such
as low pH, high cell wall binding capacity and high intracellular
metal uptake capacity (Gadd, 1987). Several fungal biosorbents
(Aspergillus niger, Aspergillus avus, Rhizopus spp., Saccharomyces
spp., and Mucor spp.) were studied as a potential biosorbent in hea-
vy metals removal from aqueous solution (Kapoor and Viraragha-
van, 1997; Kyung Kim et al., 2003; Yan and Viraraghavan, 2003;
Akar and Tunali, 2006). These fungi could remove metal ions from
aqueous solution by adsorbing metal ions on their mycelium (Bay-
ramoglu et al., 2003).
Different types of biomass (Pinus sylvestris, Azolla liculoides,
Pleurotus ostreatus, Pleurotus sajor-caju, Volvariella volvacea, Armil-
laria mellea, Daedalea quercina, Spirulina platensis, Chlorella vulgaris,
Penicillium simplicissimum, Fucus vesiculosu) have been investigated
for the biosorption characteristics of chromium(III), copper(II) and
cadmium(II) ions from aqueous solutions (Taty-Costodes et al.,
2003; Rakhshaee et al., 2006; Baldrian, 2003; Calero et al., 2009;
Pan et al., 2009; Gokhale et al., 2008; Fan et al., 2008; Mata
et al., 2009; Farooq et al., 2010).
The main concern of this work was the use of the column
packed with P. ostreatus loaded on Amberlite XAD-4 resin for the
preconcentration of Cr(III), Cd(II) and Cu(II) in aqueous solutions.
P. ostreatus was chosen in this study because it is one of the most
common edible mushrooms in Turkey and its biomass is an inex-
pensive, effective and easily cultivable biosorbent for the removal
of Cr(III), Cd(II) and Cu(II) from aqueous solutions. The metal load-
ing capacity of modied biomass was determined as a function of
pH and ow rate of sample solution, amount of solid phase, eluent
type, and concentration.
2. Methods
2.1. Microorganism and growth conditions
White-rot basidiomycete P. ostreatus was used in the experi-
ments. The stock cultures of organism were maintained on the
malt agar slants. The culture medium is a modication given by
Eaton (1985).
The culture medium contained (g L 1) 10 glucose; 0.12 NH4Cl;
2.6 K2HPO4; 0.5 MgSO4; 0.1 CaCl2; 0.001 thiamine. The pH value of
medium was 4.5. The culture was incubated at 30 C, 150 rpm for
15 days. After 15 days of growth, the harvested cells were washed
twice with deionized distilled water and in order to improve the
metal binding properties, intact cells were boiled in 0.5 N NaOH
solution for 15 min and then washed with generous amounts of
deionized distilled water till the pH of the wash solution was in
the near neutral range. Then, it was dried at 60 C for 24 h before
use.
2.2. Reagents
Analytical reagent grade CrCl3.6H2O, CdCl2.H2O, CuSO4.5H2O,
HCl, NaOH, chemicals from Merck (Germany), Cr(III), Cd(II) and
Cu(II) atomic absorption spectrometry standard solutions
(1000 mg L 1) (Fluka Chemicals) were used. Working solutions of
the metal ions were prepared by a suitable dilution of a stock solu-
tion with doubly distilled water.
Amberlite XAD-4 resin was used as a support material for the
immobilization of P. ostreatus. Amberlite XAD-4 (surface area
725 m2 g 1, pore diameter 4 nm and bead size 2060 mesh) was
supplied by Sigma. XAD-4 resin obtained from the supplier con-
tained organic and inorganic impurities. To remove the contami-
nants, it was treated with 4 M HCl. The resin was rstly rinsed
with distilled water until its pH was neutral, secondly with an eth-
anolwater (1:1) solution and nally with distilled water again.
Then, it was stored in a polyethylene bottle.
2.3. Immobilization of the microorganism onto Amberlite XAD-4
The immobilization of P. ostreatus was performed according to
the procedure recommended by Mahan and Holcombe (1992).
200 mg of dry P. ostreatus was mixed with 2 g of Amberlite XAD-
4 resin. The mixture was wetted with 2 mL of doubly distilled
deionized water and thoroughly mixed. After mixing, the paste
was heated in an oven at 80 C for 24 h to dry the mixture. The
wetting and drying step was repeated to maximize the contact be-
tween fungi and resin, thereby improving the immobilization ef-
ciency. Then, the product obtained was ground to get original size
(2040 mesh) and used as an adsorbent.
2.4. Preparation of the column
P. ostreatus immobilized resin (0.2 g) was packed in a glass col-
umn (10 mm internal diameter and 200 mm length). Before use,
1 mol L 1 HCl solution and doubly distilled deionized water were
passed through the column in order to condition and clean it. Then,
the column was conditioned to studied pH by passing an aqueous
solution of HCl or NaOH having the same pH as that of sample solu-
tion through the column, prior to passage of the sample solution.
The experiments were conducted by pumping a metal solution in
down-ow mode through the column with a peristaltic pump.
2.5. General procedure for sorption studies
An off-line column-SPE (Solid phase extraction) procedure was
applied for preconcentration. An aliquot of the sample solution
containing 2.525 lg Cr, Cd and Cu, in 100 mL was taken and the
pH was adjusted to the optimum value, determined experimen-
tally with HCl or NaOH. The samples at the chosen pH were passed
through the column at appropriate ow rates determined experi-
mentally. After passing this solution completely, the retained me-
tal ions were than eluted from the solid phase with a suitable
eluent, determined experimentally (Table 1). The concentration
of the metal ions in the eluate was determined by atomic absorp-
tion spectrometry (Analitik Jena-Nova 300).
P. ostreatus immobilized resin was used repeatedly after wash-
ing with 1 M HCl solution and than doubly distilled water.
 S. Kocaoba, M. Arsoy / Bioresource Technology 102 (2011) 80358039 8037

Table 1 100
Effect of the type and volume of elution solutions on the recovery of metals.

Element Type of elution Volume Concentration Recovery 80

solution (mL) (mol L 1) (%)

Recovery (%)
Cr(III) HCl 5 1 84.30 60
HCl 10 1 92.50
HCl 15 1 90.20 40 Cr
HNO3 5 1 78.40
Cd
HNO3 10 1 83.50 20
HNO3 15 1 84.00 Cu
Cd(II) HCl 5 1 87.30
HCl 10 1 94.70 0
0 2 4 6 8
HCl 15 1 90.30
HNO3 5 1 80.50 pH
HNO3 10 1 84.50
HNO3 15 1 87.40 Fig. 1. Effect of pH on the recovery of chromium, cadmium and copper by Pleurotus
Cu(II) HCl 5 1 95.60 ostreatus immobilized on Amberlite XAD-4 resin.
HCl 10 1 98.40
HCl 15 1 98.00
HNO3 5 1 87.50 resin, which varied from 0.05 to 0.4 g. The results are shown in
HNO3 10 1 90.10
Fig. 2. It was found that above 0.1 g of adsorbent the recovery of
HNO3 15 1 91.20
metals gradually increased, but about 0.2 g of adsorbent reached
plateau. Therefore, 0.2 g adsorbent was found to be optimum of
3. Results and discussion all preconcentration experiments.

Although there are many studies on the effects of toxic heavy
metals on microorganisms, in terms of their growth and propaga-
tion as well as their ability to accumulate these metals in living and
non-living biomass, relatively little information has been pub-
lished on the preconcentration of trace metals by using microor-
ganisms as biosorbent materials. Column procedures which are
time consuming and allow easier regeneration of the biosorbent
have been already used to immobilize fungi, yeast and algae
(Godlewska-Zylkiewicz, 2006).
In this paper, the effects of experimental parameters, such as
pH, amount of adsorbent, ow rate of sample solution, eluent type
and volume, and electrolytes and foreign ions on the recovery of
the metal ions were investigated.
3.1. The effect of pH
The pH of the sample solution plays an important role in micro-
bial biosorption. Thus the effect of the pH on the ability of the col-
umn containing P. ostreatus immobilized on Amberlite XAD-4 to
preconcentrate the metal ions was studied. The pH value of the
sample solutions was adjusted to a range of 18 with HCl or NaOH.
The obtained solutions were passed through the column at a ow
rate at about 2 mL min 1. The metal ions were then eluted by 1 M
HCl. The results are given in Fig. 1. The uptake values were in-
creased with increasing pH values for all metals. From the results
shown in Fig. 1, it could be concluded that the cell surface becomes
positively charged at low pH values which decrease the attraction
between metal ions and the functional groups on the cell wall,
whereas the cell surface becomes negatively charged at high pH
values, increasing the attraction until a maximum is reached after
pH 45. For pH values higher than the optimum values, the reten-
tion decreases again due to the competition between the hydroxyl-
ated complexes of the metal and active sites of the cell. For
3.3. The effect of ow rates of sample solutions
The ow rate of the sample solution affects the mass transfer
from the solution to the binding sites on the cell wall of microor-
ganism. For that reason, the retention of elements on an adsorbent
depends upon the ow rate of the metal solution. Therefore, the ef-
fect of the ow rate of sample solution was examined under opti-
mum conditions (pH, eluent type, etc.) by using a peristaltic pump.
The solution was passed through the column with the ow rates
adjusted in a range of 17.5 mL min 1. Flow rates slower than
1 mL min 1 were not studied to avoid long analysis times. How-
ever, at a ow rate greater than 3 mL min 1, there was a decrease
in the percentage of sorption, as the metal ions could not equili-
brate properly with the resin bed. The reason for this decrease is
probably insufcient contact of the metal ions and the adsorbent
to reach equilibrium. As shown in Fig. 3, the optimum ow rate
was found as 2.5 mL min 1 for all metal ions. Therefore,
2.5 mL min 1 was used as a ow rate for subsequent experiments.
3.4. The effect of the type and volume of elution solutions
The other important factor, that affects the preconcentration
technique, is the type and concentration of the eluent used for
the release of metal ions from the bacterial surface. The concentra-
tion of the acid used as an eluent must be the lowest possible level
in order to prevent degradation of the biomass (Bag et al., 2000).
Optimization of the elution conditions was performed in order to
100
80
Recovery (%)

avoiding precipitation of metals high pH values were not prefera- 60

ble and the optimum pH was chosen as pH 5 for all metals. Previ-
40
ous studies on heavy metal biosorption have also shown that pH Cr
was the single most important parameter affecting the biosorption 20
Cd
process (Kapoor et al., 1999; Kacar et al., 2002; Dogru et al., 2007). Cu
0
0 100 200 300 400 500
3.2. The effect of the amount of adsorbent
Adsorbent amount (mg)

The retention of the elements studied was examined in relation Fig. 2. Effect of amount of adsorbent on the recovery of chromium, cadmium and
to the amount of P. ostreatus immobilized on Amberlite XAD-4 copper by Pleurotus ostreatus immobilized on Amberlite XAD-4 resin.
8038 S. Kocaoba, M. Arsoy / Bioresource Technology 102 (2011) 80358039

100 Table 2
Precision of the method.

Element Recoverya (%) N

90
Recovery (%)

Cr 98.6 0.4 5
Cd 97.5 0.3 5
80 Cu 95.8 0.4 5
a
Cr Average of N determinations with 95% condence level.
70 Cd
Cu
Amberlite XAD-4 loaded with P. ostreatus several times using a
60
0 2 4 6 8 solution (50 mL) having a concentration of 550 lg mL 1 under
Flow rate (mLmin-1) optimum experimental conditions. It was found that the sorption
capacity after 10 cycles of sorption and desorption does not vary
Fig. 3. Effect of ow rate on the recovery of chromium, cadmium and copper by
more than 2.0%. The columns seem to be relatively stable up to
Pleurotus ostreatus immobilized on Amberlite XAD-4 resin.
10 cycles. Therefore, repeated use of the resin is possible.

obtain the maximum recovery with the minimal concentration and
volume of the elution solution. The effect of different volumes of
nitric acid and hydrochloric acid in water was tested to remove
the bound metal ions from the bacterial biomass loaded onto the
column. The elution studies were performed with 1 mol L 1 HCl
and HNO3 solutions. The eluate volume was 5, 10 and 15 mL. As
can be seen from Table 1 and 10 mL of 1 mol L 1 HCl solution
was found to be satisfactory for all elements studied.
3.7. Precision of the method
For evaluation of the precision of the method, the optimum con-
ditions mentioned above were used, and successive retention and
elution cycles (with 25 lg of Cr, Cd and Cu) were performed. As
can be seen in Table 2, the precision of the method is very good,
and the recoveries of analytes are quantitative (above 95%).

3.8. Effect of electrolytes and foreign ions

3.5. Effect of the volume of sample solutions
The effect of changes in the volume of sample solution passed
through the column on the recovery of Cr(III), Cd(II) and Cu(II)
was investigated in order to determine an applicable sample vol-
ume or a minimum analyte concentration. For this purpose, 50,
100, 250, 500, 750 and 1000 mL volume solution containing 0.25,
0.10, 0.05, 0.033 and 0.025 lg mL 1 of Cr, Cd and Cu were passed
through the column under the optimum conditions. It was found
that all metal ions could be recovered quantitatively (P95%) up
to 250 mL of the sample solution (Fig. 4). At the higher volumes,
the recoveries for analytes decreased. In this study, the elution vol-
ume was 10 mL, therefore the preconcentration factors were 25 for
the analytes. These results show that Cr, Cd and Cu could be deter-
mined in low concentrations by the proposed method, which could
not be determined directly by AAS.
One of the main problem in the atomic absorption spectromet-
ric determination of the heavy metal ions is interference from the
matrix. In this study, the inuences of the some ions such as NaCl,
KBr, KI, NaNO3, Na2SO4, Na3PO4, Ca(II) and Mg(II), which are
known as interferic ions in the AAS determination, on the sorption
of Cr(III), Cd(II) and Cu(II) metal ions by P. ostreatus loaded onto
Amberlite XAD-4 in the column were investigated. The tolerance
limits of the electrolytes or cations are given in Table 3. The re-
ported tolerance limit is dened as the electrolyte ion concentra-
tion causing an error of 5%. These results show that the ions
present in water at their normal concentration levels do not inter-
fere under the experimental conditions used. It means that the pro-
posed preconcentration method could be applied to natural water
samples that contain such ions at the tolerable levels given in Table
3.

3.6. The effect of column reuse 3.9. Application

The stability and potential recyclability of the column were as- The proposed method was applied to the determination of chro-
sessed by monitoring the change in the recoveries of metal ions mium, cadmium and copper in tap water, employing standard
through several adsorptionelution cycles. In order to study these addition. An appropriate volume of sample solutions was adjusted
effects, the metal ions were sorbed and desorbed on 200 mg of the to the optimum pH and subjected to the recommended column
procedure for the preconcentration and determination of metal
ions.
100
The results reported in Table 4, with a condence interval for
Cr the 95% condence level, show the applicability of the proposed
95
Cd method to water analysis. The analytes were determined with a
Recovery (%)

Cu relative error lower than 10% in all samples. When the results com-
90
pared with permissible limits of these metals in tap water, it shows
85

Table 3
80 Tolerance limits of electrolytes on the sorption of metal ions on resin.
1
Metal Electrolytes or metal ions (mol L )
75
0 250 500 750 1000 NaCl KBr KI NaNO3 Na2SO4 Na3PO4 Ca(II) Mg(II)
Volume of sample solution (mL) Cr(III) 0.15 0.15 0.12 0.10 0.07 0.05 0.12 0.12
Cd(II) 0.15 0.12 0.10 0.05 0.05 0.05 0.10 0.12
Fig. 4. Effect of volume of sample solutions on the recovery of chromium, cadmium Cu(II) 0.15 0.13 0.10 0.10 0.06 0.05 0.12 0.10
and copper by Pleurotus ostreatus immobilized on Amberlite XAD-4 resin.
 S. Kocaoba, M. Arsoy / Bioresource Technology 102 (2011) 80358039 8039

Table 4 Calero, M., Hernainz, F., Blazquez, G., Tenorio, G., Martin-Lara, M.A., 2009. Study of
Determination of chromium, cadmium and copper in tap Cr (III) biosorption in a xed-bed column. J. Hazard. Mater. 171, 886893.
water. Davis, T., Volesky, B., Mucci, A., 2003. A review of the biochemistry of heavy metal
biosorption by brown algae. Water Res. 37, 43114330.
Element Added (lg) Recoveryb (%) Dogru, M., Guven, R., Erdogan, S., 2007. The use of Bacillus subtilis immobilized on
Amberlite XAD-4 as a new biosorbent in trace metal determination. J. Hazard.
Cr 10 96.5 0.4
Mater. 149, 166173.
20 97.3 0.4 Eaton, D.C., 1985. Mineralization of polychlorinated-biphenyls by Phanerochaete-
30 98.4 0.3 Chrysosporium-A ligninolytic fungus. Enzym. Microb. Technol. 7 (5), 194196.
Cd 10 95.4 0.4 Fan, T., Liu, Y., Feng, B., Zeng, G., Yang, C., Zhou, M., Zhou, H., Tan, Z., Wang, X., 2008.
20 96.6 0.3 Biosorption of cadmium(II), zinc(II) and lead(II) by Penicillium simplicissimum:
30 97.5 0.3 isotherms, kinetics and thermodynamics. J. Hazard. Mater. 160, 655661.
Cu 10 93.8 0.4 Farooq, U., Kozinski, J.A., Khan, M.A., Athar, M., 2010. Biosorption of heavy metal
20 94.6 0.4 ions using wheat based biosorbents a review of the recent literature.
30 95.2 0.3 Bioresource Technol. 101, 50435053.
Gadd, G.M., 1987. Fungal Response Towards Heavy Metals. In: Herbert, R.A., Codd,
b
Mean of 5 determinations with 95% condence level. G.A. (Eds.), Microbes in Extreme Environmentals. Academic Press, London U.K.
Godlewska-Zylkiewicz, B., 2006. Microorganisms in inorganic chemical analysis.
Anal. Bioanal. Chem. 384, 114123.
that the method is very convenient for determination of Cr(III), Gokhale, S.V., Jyoti, K.K., Lele, S.S., 2008. Kinetic and equilibrium modeling of
Cd(II) and Cu(II). chromium (VI) biosorption on fresh and spent Spirulina platensis/Chlorella
vulgaris biomass. Bioresource Technol. 99, 36003608.
Gong, R., Ding, Y.D., Liu, H., Chen, Q., Liu, Z., 2005. Lead biosorption by intact and
pretreated Spirulina maxima biomass. Chemosphere 58, 125130.
4. Conclusion Holan, Z.R., Volesky, B., Prasetyo, I., 1993. Biosorption of cadmium by biomass of
marine algae. Biotechnol. Bioeng. 41, 819825.
The results presented in this paper demonstrate the usability of Holan, Z.R., Volesky, B., 1995. Accumulation of cadmium, lead, and nickel by fungal
and wood biosorbents. Appl. Biochem. Biotechnol. 53, 133146.
a new biosorbent, P. ostreatus, for the preconcentration of heavy Kacar, Y., Arpa, C., Tan, S., Denizli, A., Genc, O., Arca, M.Y., 2002. Biosorption of Hg(II)
metals. The biomass, with higher metal sorption capacity and and Cd(II) from aqueous solutions: comparison of biosorptive capacity of
mechanical strength after immobilized on Amberlite XAD-4, may alginate and immobilized live and heat inactivated Phanerochaete
chrysosporium. Process Biochem. 37, 601610.
nd potential application in wastewater treatment for metal ions Kapoor, A., Viraraghavan, T., 1997. Heavy metal biosorption sites in Aspergillus niger.
removal. The analytes could be preconcentrated directly by using Bioresource Technol. 61, 221227.
the proposed method without using any chelating or complexing Kapoor, A., Viraraghavan, T., Cullimore, D.R., 1999. Removal of heavy metals using
the fungus Aspergillus niger. Bioresource Technol. 70 (1), 95104.
agent which usually used to increase retention. The preconcentra-
Kocaoba, S., 2007. Comparison of Amberlite IR 120 and dolomites performances for
tion technique used in this work is also cheap. Only 0.2 g of adsor- removal of heavy metals. J. Hazard. Mater. 147, 488496.
bent is needed and repeated use is possible. The optimum pH was Krishna, K.R., Philip, L., 2005. Bioremediation of Cr(VI) in contaminated soils. J.
Hazard. Mater. 121, 109117.
chosen as pH 5 for all metals. The optimum ow rate was found as
Kyung Kim, S., Park, C.B., Koo, Y.M., Yun, H.S., 2003. Biosorption of cadmium and
2.5 mL min 1 for all metal ions. The effect of different volumes of copper ions by Tricoderma reesei RUT C30. J. Ind. Eng. Chem. 9, 403406.
nitric acid and hydrochloric acid in water was tested to remove Mahan, C.A., Holcombe, J.A., 1992. Immobilization of algae cells on silica gel and
the bound metal ions and 10 mL of 1 mol L 1 HCl solution was their characterization for trace metal preconcentration. Anal. Chem. 64 (17),
19331939.
found to be satisfactory for all elements studied. It was found that Mata, Y.N., Blazquez, M.L., Ballester, A., Gonzalez, F., Munoz, J.A., 2009. Biosorption
all metal ions could be recovered quantitatively (P95%) up to of cadmium, lead and copper with calcium alginate xerogels and immobilized
250 mL of the sample solution. The procedure shows improved Fucus vesiculosus. J. Hazard. Mater. 163, 555562.
Pan, R., Cao, L.X., Zhang, R.D., 2009. Combined effects of Cu, Cd, Pb, and Zn on the
retention of analytes, better repeatability, and multiple applica- growth and uptake of consortium of Cu-resistant Penicillium sp. A1 and Cd-
tions possibilities of the column (about 10 runs). The matrix effects resistant Fusarium sp. A19. J. Hazard Mater. 171, 761766.
were reasonably tolerable. Pardo, R., Herguedas, M., Barrado, E., Vega, M., 2003. Biosorption of cadmium,
copper, lead and zinc by inactive biomass of Pseudomonas putida. Anal. Bioanal.
In conclusion, the proposed method is excellent as regards sim- Chem. 376, 2632.
plicity, sensitivity, selectivity, precision, accuracy and column Pavasant, P., Apiratikul, R., Sungkhum, V., Suthiparinyanont, P., Wattanachira, S.,
stability. Marhaba, T.F., 2006. Biosorption of Cu2+, Cd2+, Pb2+, and Zn2+ using dried marine
green macroalga Caulerpa lentillifera. Bioresource Technol. 97, 23212329.
Prasad, M.N.V., Freitas, H., 2000. Removal of toxic metals from solution by leaf, stem
References and root phytomass of Quercus ilex L. (holly oak). Environ. Pollut. 110, 277283.
Rakhshaee, R., Khosravi, M., Ganji, M.T., 2006. Kinetic modeling and thermodynamic
study to remove Pb(II), Cd(II), Ni(II) and Zn(II) from aqueous solution using dead
Abu Al-Rub, F.A., El-Naas, M.H., Benyahia, F., Ashour, I., 2004. Biosorption of nickel
and living Azolla liculoides. J. Hazard. Mater. 134, 120129.
on blank alginate beads, free and immobilized algal cells. Process Biochem. 39,
Saeed, A., Iqbal, M., Akhtar, M.W., 2005. Removal and recovery of lead(II) from single
17671773.
and multimetal (Cd, Cu, Ni, Zn) solutions by crop milling waste (black gram
Akar, T., Tunali, S., 2006. Biosorption characteristics of Aspergillus avus biomass for
husk). J. Hazard. Mater. 117, 6573.
removal of Pb(II) and Cu(II) ions from an aqueous solution. Bioresource Technol.
Taty-Costodes, C.V., Fauduet, H., Porte, C., Delacroix, A., 2003. Removal of Cd(II) and
97, 17801787.
Pb(II) ions, from aqueous solutions, by adsorption onto sawdust of Pinus
Bag, H., Turker, A.R., Lale, M., 2000. Determination of Cu, Zn, Fe, Ni and Cd by ame
sylvestris. J. Hazard. Mater. 105, 121142.
atomic absorption spectrophotometry after preconcentration by Escherichia coli
Tsezos, M., 2001. Biosorption of metals. The experience accumulated and the
immobilized on sepiolite. Talanta 51, 10351043.
outlook for technology development. Hydrometallurgy 59, 241243.
Baldrian, P., 2003. Interactions of heavy metals with white-rot fungie. Enzyme
Xu, H., Liu, Y., 2008. Mechanisms of Cd2+, Cu2+ and Ni2+ biosorption by aerobic
Microb. Technol. 32, 7891.
granules. Sep. Purif. Technol. 58, 400411.
Baral, A., Engelken, R.D., 2002. Chromium-based regulation and greening in metal
Yan, G., Viraraghavan, T., 2003. Heavy metal removal from aqueous solution by
nishing industries in the USA. Environ. Sci. Pollut. 5, 121133.
fungus Mucor rouxii. Water Res. 37, 44864496.
Bayramoglu, G., Bektas, S., Arica, M.Y., 2003. Biosorption of heavy metals on _
Yaramaz, O., 1992. evre ve Su Kirliligi Ege niversitesi, Su rnleri Fakltesi. Izmir.
immobilized white-rot fungus Trametes versicolor. J. Hazard. Mater. 101, 285
42, 192.
300.