Bioresource Technology 97 (2006) 512–521

Biosorption of dyes using dead macro fungi:

Effect of dye structure, ionic strength and pH
Nityanand Singh Maurya a, Atul Kumar Mittal a,*
, Peter Cornel b, Elmar Rother b

a
Department of Civil Engineering, Indian Institute of Technology Delhi, New Delhi 110 016, India
b
Institute WAR, Technische Universität Darmstadt, Petersenstraße 13, 64287 Darmstadt, Germany

Received 15 January 2004; received in revised form 13 December 2004; accepted 28 February 2005
Available online 20 April 2005

Abstract

Biosorbents prepared from dead macro fungi, namely Fomes fomentarius and Phellinus igniarius, were applied for the uptake of
Methylene Blue (MB) and Rhodamine B (RB). Equilibrium isotherm data could be well described by the Langmuir and Freundlich
models. Methylene Blue was found to be more adsorbable than Rhodamine B. Langmuir monolayer coverage was determined as
204.38–232.73 mg/g and 25.12–36.82 mg/g for MB and RB, respectively. Molecular structure and ionic radius of dyes were found to
be responsible for differences in their uptakes. Results showed that sorption of MB increased while that of RB decreased as pH of
respective dye solutions changed from 3 to 11. An increase in ionic strength also exhibited an adverse effect on dye sorption capacity.
Ionic strength and pH affected the sorption of MB more as compared to the sorption of RB. The presence of carboxylic (
ve) and
amino (+ve) groups in RB could explain the lower sorption of RB compared to MB.

2005 Elsevier Ltd. All rights reserved.

Keywords: Macro fungus; Biosorption; Cationic dye; Dye structure; Ionic strength and pH

1. Introduction food industries, pulp and paper, paint and electroplating

Colour is a visible pollution. Even a slight colour-
ation of water sources could make them unacceptable
to consumers though it may not be toxic to the same de-
gree. The source of such pollution lies in the rapid in-
crease in the use of synthetic dyes. More than 10,000
chemically different dyes are being manufactured. The
world dyestuff and dye intermediates production is esti-
mated to be around 7 · 108 kg per annum (Fu and
Viraraghavan, 2001; Toh et al., 2003). These are mainly
consumed in textiles, tanneries, pharmaceuticals, packed
*
Corresponding author. Tel.: +91 11 2659 1239; fax: +91 11 2658
1117.
E-mail address: akmittal@civil.iitd.ernet.in (A.K. Mittal).
industries. The effluents from industries, which include
both dye manufacturing and dye application, are highly
coloured. Besides dyes, such effluents contain a number
of other contaminants such as acids or alkalis, salts,
dissolved and suspended solids, and other toxic
compounds. Colour acquired by receiving water bodies
such as rivers or lakes inhibits growth of the desirable
aquatic life necessary for self-purification by reducing
penetration of sunlight, with a consequent reduction in
photosynthetic activity. Water treatment cost of the util-
ities located downstream of coloured water bodies is
high. Colour causing compounds can react with metal
ions to form substances which are very toxic to fish
and other aquatic life (Karthikeyan, 1989).
The majority of technologies presently employed for
colour removal are based on physicochemical processes

0960-8524/$ - see front matter
2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2005.02.045
 N.S. Maurya et al. / Bioresource Technology 97 (2006) 512–521
such as dilution, adsorption, coagulation and floccula-
tion, chemical precipitation, oxidation, ion-exchange,
reverse osmosis and ultra filtration (Banat et al., 1996).
The other class of processes is based on the biodegrad-
ability of dyes. Generally, dyes are difficult to
biodegrade (Fu and Viraraghavan, 2000). Application
of biological processes to treat coloured wastewater is
yet to be fully established. Among treatment technol-
ogies, adsorption is rapidly gaining prominence as a
method of treating aqueous effluent. Activated carbon
is the most effective and widely used adsorbent (El-
Geundi, 1991). However, its high cost has prevented
its application, at least in developing countries, so it is
necessary to search for an alternative low-cost adsorbent
which can be used as a substitute for activated carbon. A
number of low cost adsorbents are reported in the
literature. These include bagasse pith (Nassar and El-
Geundi, 1991; McKay et al., 1988), maize cobs
(El-Geundi, 1991), sunflower (Sun and Xu, 1997), fly
ash (Gupta et al., 2000), peat (Ramakrishna and Vira-
raghavan, 1996) and saw dust (El-Nabarwy and Khedr,
2000). However, sorption potential of these low cost
sorbents is generally low. Since regeneration of these
sorbents is generally not possible, a large quantity of
spent sorbents need to be disposed of, which causes
the waste disposal problem of dye-bearing sorbents.
Therefore, there is a need to find an economical, easily
available (in nature) and highly effective sorbent, which
can be easily regenerated.
Biosorption can be defined as sequestering of organic
and inorganic species including metals, dyes and odor
causing substances using live or dead biomass or their
derivatives. This biomass may be bacteria, fungi, algae,
sludge from biological wastewater treatment plants, by-
products from fermentation industries or seaweeds. In
this process, adsorbents are biological materials, and
the removal mechanism is mainly sorption. Since the
1980s, biosorption has been continuously studied for
the removal of heavy metals and other pollutants from
wastewater, so it could be a promising alternative to
replace or supplement present dye bearing wastewater
treatment processes. Biosorption, if compared with
other available technologies such as precipitation, ion
exchange, reverse-osmosis and adsorption, gives compa-
rable performance at a very low cost. Apart from cost
effectiveness and competitive performance, other advan-
tages are possible regeneration at low cost, availability
of known process equipment, sludge free operation
and recovery of the sorbate (Volesky, 1999).
The biosorption capacity of a biomass depends on
several factors. It includes type of biomass (species,
age), type of sorbates, presence of other competing ions
and method of biomass preparation (culture condition
for live biomass), along with several physico-chemical
factors (temperature, pH, ionic concentration). In
applying biosorption as a process in an industrial envi-
513
ronment, a choice has to be made between live and dead
biomass. The use of dead biomass appears to be more
attractive compared with the live biomass, because the
toxic nature of ions or pollutants cannot have any effect
on the sorption process. Besides, there is no requirement
for either having any nutrients or maintaining a growth
environment. Apart from these factors, researchers have
also reported that dead biomass is more effective in
adsorbing organic pollutants than live biomass (Tsezos
and Bell, 1989; Hong et al., 2000). Other studies with
some organic compounds have also suggested that the
fluidity of the membrane that can be affected by temper-
ature could be one of the factors determining the
adsorption ratio of the organic pollutants (Wiegert
and Harrie, 1995).
Over the past few decades, a number of biological
materials have been studied for the sorption of different
types of pollutants, including heavy metals and organic
materials. Literature shows that a wide variety of micro-
organisms, including bacteria, algae and fungi, are cap-
able of sorbing a number of pollutants (Ju et al., 1997;
Fu and Viraraghavan, 2000; Walker and Weatherley,
2000; Schiewer and Wong, 2000). Among these micro-
organisms, fungal biomass seems to be a good sorption
material, because it can be produced economically using
simple fermentation techniques and economical growth
media. Fungal biomass is also available as a by-product
or waste material from various industrial processes (Fu
and Viraraghavan, 2000).
Among the commonly available sorbents, macro fun-
gus possesses the characteristics of an ideal biosorbent.
It is easily and economically available anywhere, espe-
cially in locations with hot and humid climates such as
India. A macro fungus-based biosorbent is chemically
stable in most alkaline and acidic conditions. It also pos-
sesses good mechanical properties against abrasion
(Mittal and Gupta, 1996).
In the present study, two different types of dead
macro fungi, which were collected from forests in Ger-
many (Atleiningen and Darmstadt), were used. Methy-
lene Blue and Rhodamine B were selected as the
sample cationic dyes to evaluate the sorption potential
of the biosorbents. These dyes, compared to other dyes,
have high brilliance and colour intensity, resulting in
high visible colour, even at a very low dye concentration
(Fu and Viraraghavan, 2000). Besides, sorption of these
dyes has been reported by other researchers using differ-
ent sorbents (Kannan and Sundaram, 2001; Namasiva-
yam et al., 2001; Miguel et al., 2002; Annadurai et al.,
2002; Fu and Viraraghavan, 2002; Rozada et al., 2003;
Reyad and Maha, 2003; Pavel et al., 2003). So, the sorp-
tion parameters obtained using the present biosorbents
could readily be compared with the ones presented in
the literature. The effects of pH and ionic strength were
also investigated, as well as the sorption mechanism of
dye uptake.
514 N.S. Maurya et al. / Bioresource Technology 97 (2006) 512–521
2. Methods
2.1. Materials
2.1.1. Sorbents
The biosorbents used in the study were prepared from
dead macro fungi (mushroom). These mushrooms (Phel-
linus igniarius and Fomes fomentarius) were collected
from forests (Atleiningen and Darmstadt, Germany).
The biosorbent prepared from Phellinus igniarius is
abbreviated as ‘‘AM’’ and that from Fomes fomentarius
is ‘‘BM’’.
The macro fungi were picked manually, washed in
water, cut into small pieces and dried in an oven at 40
C for 48 h. The dried adsorbents were pulverised in
an electric ball mill. The pulverised sorbents were passed
through a 600 lm sieve to discard bigger particles. The
sieved adsorbents were kept in an oven at 40 C for 24
h and then stored in a desiccator without any further
chemical or physical treatment.
2.1.2. Adsorbates
Two cationic dyes, namely Methylene Blue (MB) and
Rhodamine B (RB) were selected as the representa-
tive cationic dyes. MB is the representative of a wholly
basic cationic dye group, whereas RB is the representa-
tive of cationic dye having acidic colligator. RB can be
assumed as the connection between basic and acid dye.
Aqueous dye solutions of 100 and 200 mg/L were pre-
pared in extraction water. The extraction water was pre-
pared using ultra pure (Millipore) water and 0.5, 0.3 and
0.2 mmol/l NaHCO3, CaCl2 Æ 2H2O and MgSO4 Æ 7H2O,
respectively. The pH of extraction water was adjusted to
7.5 ± 0.2. Information on dyes is given in Table 1.
2.2. Methods
2.2.1. Measurement of dye concentration
Dye concentration was measured spectrophotometri-
cally using a PMQ3 Spectrophotometer (Zeiss, West
Germany). All absorbance determinations were carried
out at the wavelengths corresponding to the respective
maximum absorbance, kmax, and at 10 mm optical path
Table 1
Dyes employed in the study
Commercial name Abbreviation Application class Chemical structure
Methylene blue MB Basic blue 9
Rhodamine B RB Basic violet 10
length. Measurements were conducted in the optical
density range of 0.080–0.230.
2.2.2. Equilibrium studies
Bottle point isotherm experiments were conducted
using the two biosorbents AM and BM, and two sor-
bates, MB and RB. Fifty millilitres aqueous dye solu-
tions were added to 100 mL bottles and different doses
(0.07–2.10 g/L for MB, 0.20–7.80 g/L for RB) of sorbent
were added to the dye solution. All bottles were loaded
on an end-to-end rotary shaker. Experiments were con-
ducted in duplicate with two controls (having no biosor-
bent). Controls were employed to ensure that sorption
was by fungal biomass only, and any effect of sorption
of dye onto the wall of the reaction bottles could be
ruled out. Bottles were rotated at 30 rpm for 48 h at
20 C. Upon equilibration, the shaking was stopped,
and samples from each bottle were withdrawn with a
glass syringe. The dye solution was separated from the
sorbent by filtering through a 0.45 lm membrane filter
(Schleicher and Schuell, Germany). The effect of dye
sorption by the membrane filter during the filtration
process was minimised by discarding the first 25 mL
portion of the dye solution. The subsequent portion
was used for the measurement of absorbance for the
determination of dye concentration. Dye control solu-
tions (dye solution without sorbent) were also filtered
using the same procedure and the dye concentration
was determined using a spectrophotometer. The dye
concentration of the control was used as the initial con-
centration for calculating the quantity of dye transferred
from the dye solution to the sorbent.
2.2.3. Effect of ionic strength
Aqueous dye solutions of 100 mg/L dye concen-
tration were prepared. Ionic strengths of these dye
solutions were varied using NaCl, CaCl2 Æ 2H2O and
MgCl2 Æ 6H2O. Fifty millilitres dye solutions having dif-
ferent ionic strengths (<0.001, 0.1, 1, 10 and 100 mmol/l)
were added to 100 mL bottles and were contacted with
biosorbents for 48 h. Equilibrium dye concentrations
were measured using the spectrophotometer after sepa-
ration of sorbent from the dye-sorbent mixture.
Color index Molecular wt. kmax (nm) Chromopore
52015 319.86 665 Thiazine
group
45170 479.03 555 Xanthene
group
 N.S. Maurya et al. / Bioresource Technology 97 (2006) 512–521 515

All the experiments related to the effect of ionic
strength were carried out in duplicate and mean values
are presented. Maximum deviation was ±1%.
2.2.4. Effect of pH
The effect of pH on dye uptake was evaluated by con-
tacting a predetermined amount of biosorbent (0.3 g/L
for MB and 0.8 g/L for RB) with dye solutions at differ-
ent initial pH (3, 5, 7.5, 9 and 11), keeping other param-
eters constant. No buffer was used to adjust the pH since
a single buffer could not effectively cover the whole
range. Furthermore, different components used to make
various buffers may themselves affect the sorption to dif-
fering extents. Fifty millilitres dye solution with 100-mg/
L concentration was added to a 100 mL bottle and pH
was varied using 0.1N HCl/NaOH. An equal amount
of sorbent (0.3 g/L for MB and 0.8 g/L for RB) was
added to each sample and contacted for 48 h on a rotary
shaker at 20 C. Initial and equilibrium pH of dye solu-
tions were measured.
3. Results and discussion
3.1. Biosorption isotherms
Biosorption data are most commonly represented by
an equilibrium isotherm, which is a plot of the quantity
of sorbate retained on biosorbent Q (solid phase concen-
tration of the sorbate) as a function of the concentration
of the sorbate in the liquid phase Ce (liquid phase concen-
tration of the sorbate at equilibrium). At equilibrium, a
dynamic equilibrium is established in the concentration
of sorbate between two phases. The equilibrium isotherm
is of fundamental importance for the design and optimi-
zation of the adsorption system for the removal of dye
from aqueous solution. Therefore, it is necessary to estab-
lish the most appropriate correlation for the equilibrium
curves. Presently several isotherm models are available.
In the present study, two of the most commonly used
models, namely Langmuir and Freundlich isotherms,
were used to describe the biosorption equilibrium.
The Langmuir isotherm model is an analytical equa-
tion basically developed for gas phase adsorption onto
the homogeneous surface of glass and metals. This
model is one of the ideal limiting-condition type models.
It is predicted on a uniform surface affinity for the solute
and prescribes a non-linear asymptotic approach to a
single maximum sorption capacity (Langmuir, 1918;
Weber Jr., 1972; Faust and Aly, 1987). The Freundlich
isotherm model is an empirical expression that encom-
passes the heterogeneity of the surface and the exponen-
tial distribution of sites and their energies (Faust and
Aly, 1987). It is the most widely used mathematical
description of sorption in aqueous-solid systems. The
Langmuir model has the form:
Qm bC e
Q¼ ð1Þ
1 þ bC e
The linear form of the Langmuir equation (1) is
     
Ce Ce 1
¼ þ ð2Þ
Q Qm bQm
The Freundlich model can be written as:
1
Q ¼ KC ne ð3Þ
Its linear form is given below:
1
log Q ¼ log K þ log C e ð4Þ
n
Here, Q is solid phase sorbate concentration (mg/g), Ce
is the aqueous phase sorbate concentration (mg/L), Qm
and b are the Langmuir model constants and K and n
are the Freundlich model constants.
For the determination of model constants, the Lang-
muir and Freundlich models could be analyzed using
linear forms as described in Eqs. (2) and (4). However,
determination of model constants using these linearised
forms of the isotherm models could be erroneous. Such
data transformations may alter the error structure and
could also violate the error variance and normality
assumptions of standard least squares (Myers, 1990;
Ratkowski, 1990; Allen et al., 2003). Therefore, in the
present study, model constants were determined using
non-linear regression analysis of the batch biosorption
data. Table 2 shows the model constants along with cor-
relation coefficients for biosorption of MB and RB onto
AM and BM. The comparisons between experimen-
tal data and theoretical plots of Langmuir and Freund-
lich isotherm models are shown in Figs. 1–4. For the

Table 2
Isotherm model constants
Biosorbent Dyes Langmuir model Freundlich model
2
Qm b R K n R2
Average 95% (±) Average 95% (±) Average 95% (±) Average 95% (±)
AM MB 204.38 7.13 0.184 0.033 0.987 72.26 12.54 4.472 0.863 0.952
RB 36.82 4.01 0.027 0.007 0.975 3.66 1.18 2.270 0.403 0.961
BM MB 232.73 21.47 0.140 0.054 0.974 57.00 7.26 3.129 0.329 0.992
RB 25.12 1.29 0.050 0.009 0.994 4.90 1.05 3.080 0.505 0.986
516 N.S. Maurya et al. / Bioresource Technology 97 (2006) 512–521

250 30

200
25
Qe (mg/g)

150
20
Expt Data (AM)
100

Qe (mg/g)
Langmuir (AM)
50 15
Freundlich (AM)
Expt Data (AM)
0 10
0 50 100 150 200 Langmuir (AM)
Ce (mg/L) Freundlich (AM)
5
Fig. 1. Biosorption of MB onto AM at 20 C, biosorbent dose
0.07–2.10 g/L.
0
0 20 40 60 80 100
300 Ce (mg/L)

250 Fig. 3. Biosorption of RB onto AM at 20 C, biosorbent dose

0.20–7.80 g/L.
200
Qe (mg/g)

150 25
Expt Data (BM)
100 Langmuir (BM) 20

50
Freundlich (BM)
Qe (mg/g) 15

10 Expt Data (BM)

0
0 20 40 60 80 100 Langmuir (BM)
Ce (mg/L) 5 Freundlich (BM)

Fig. 2. Biosorption of MB onto BM at 20 C, biosorbent dose 0

0 20 40 60 80 100
0.07–2.10 g/L.
Ce (mg/L)

Fig. 4. Biosorption of RB onto BM at 20 C, biosorbent dose

Langmuir model, correlation coefficient, R2 (Table 2)
varied from 0.974 to 0.994, while for Freundlich, it var-
ied from 0.952 to 0.992.
Students t-tests were also performed between experi-
mental data and data predicted using the Langmuir and
Freundlich models. The Paired or Dependent Students
test was used since the same data set was employed. Sta-
tistically, both models provided significant correlations
(at 95%, p > 0.05) as shown in Table 3. The last column
of Table 3 shows the probability of significance of the
students t-test. For a t-test to be significant, the proba-
bility should be less than 0.05. However, in all cases, it
0.20–7.80 g/L.
was more than 0.05, which indicated that the observed
values and predicted values were not significantly
different.
Both the Langmuir and the Freundlich models pro-
vided good correlations, since all the correlation coeffi-
cients were more than 0.95. However, there was a
slight difference between different correlation coeffi-
cients. Students t-test is used to check whether differ-
ent values of coefficient of correlations obtained are

Table 3
Statistical significance for the experimental and model predicted isotherm data
Biosorbent Dye Model Q t df Sig (2 tailed)
Std. deviation 95% Confidence interval
of the difference
Lower Upper
AM MB Langmuir 6.5430
2.2806 3.2451 0.361 23 0.721
AM MB Frenudlich 12.3711
5.6783 4.7694
0.180 23 0.859
BM MB Langmuir 12.2642
6.2679 10.2104 0.533 10 0.606
BM MB Freundlich 6.9111
5.0128 4.2732
0.177 10 0.863
AM RB Langmuir 1.0634
0.5876 0.4700
0.235 17 0.817
AM RB Freundlich 1.3323
0.6737 0.6513
0.036 17 0.972
BM RB Langmuir 0.4479
0.2934 0.2479
0.183 12 0.858
BM RB Freundlich 0.6708
0.4284 0.3823
0.124 12 0.903
 N.S. Maurya et al. / Bioresource Technology 97 (2006) 512–521
Table 4
Statistical significance for the isotherm data predicted by Langmuir and Freundlich isotherms
Biosorbent Dye Q t
Std. deviation 95% Confidence interval
of the difference
Lower
AM MB 16.3155
7.8262
BM MB 16.4745
13.4088
AM RB 0.5780
0.2398
BM RB 0.4689
0.2837
significantly different or not. Students t-test (paired or
dependent) was performed using the Q values predicted
by the two models, i.e., the Langmuir and Freundlich
corresponding to the same Ce value. Table 4 presents
the t-test analysis. It is evident that for all systems that
there was no significant difference between the Q values
predicted by the Langmuir and the Freundlich models
(at 95%, p > 0.05).
3.1.1. Quantification of the equilibrium isotherm data
The model constants obtained from the isotherm
models, along with their correlation coefficients (R2),
are presented in Table 2. Both the Langmuir and Fre-
undlich models provided a good fit (R2 > 0.975 for the
Langmuir model and R2 > 0.952 for the Freundlich
model) for all the sorbate and sorbent systems used in
the study. The high correlation coefficients showed the
applicability of both models for describing the sorption
equilibrium. The Langmuir and Freundlich constants
can be used to compare the sorption capacity of sor-
bents for MB and RB. For MB, Langmuir sorption
capacities, Qm, also known as monolayer sorption
capacity of sorbent, were 204.38 and 232.73 mg/g for
AM and BM biosorbents, respectively. Varying values
of the Langmuir sorption capacity Qm (20–340 mg/g)
for different sorbents have been reported in the litera-
ture. For dead fungal biomass A. niger the reported
sorption capacity is 54.67 mg/g (Fu and Viraraghavan,
2000). Sun and Xu (1997) have reported Qm for Sun-
flower stalk as 205.4 mg/g. Lee et al. (1997) have re-
ported the Qm values as 94.5, 99.0, 120.5, 128.9, 198.0
and 240.0 mg/g for the uptake of MB by carbonized
spent bleaching earth, coconut husk, hexane-extracted
spent bleaching earth, water hyacinth root, hydrilla ver-
ticillata and cotton waste, respectively. The Qm values
obtained for MB are comparable with those reported
in the literature. High Qm values (304.6–396.57 mg/g)
for chitin have been reported in the literature (Anna-
durai and Krishnan, 1996). However, chitin is obtained
after chemical and physical processing of various mate-
rials, while the present biosorbent is employed without
any chemical processing.
The monolayer sorption capacities for RB were
36.82 and 25.12 mg/g for AM and BM, respectively.
517
df Sig (2 tailed)
Upper
5.9527
0.281 23 0.781
8.7267
0.471 10 0.648
0.3350 0.349 17 0.731
0.2830
0.003 12 0.998
The reported biosorption capacities of orange peel
(Namasivayam et al., 1996), banana pith (Namasivayam
et al., 1993), banana peel (Annadurai et al., 2002) and
coir pith (Namasivayam et al., 2001) for the uptake of
RB are 3.2, 8.5, 20.6 and 203.2 mg/g, respectively. The
biosorbents investigated in the present study provided
higher sorption capacities than all of these except for
the coir pith.
MB is more adsorbable than RB. Both MB and RB
are chloride salts, but they have different molecular
structures (Table 1). MB contains three benzene rings
in a linear structure having no side chain, while RB con-
tains four benzene rings having one side chain with
acidic colligator, i.e. a carboxylic group (–COOH).
Thus, MB is grouped as a cationic dye having wholly
basic nature, while RB is known as a cationic dye having
acidic colligator (Gurr, 1971). RB is a more aromatic
compound than MB and contains a carboxylic group
(–COOH), which has pKa equal to 4.2. As pH is raised
above 4.2, there is a decrease in the overall charge of
RB. Thus at pH 7.5, at which isotherm experiments were
carried out, RB was almost neutral, whereas MB carried
a net positive charge. From the present results, it seems
that the presence of a carboxyl group, a higher number
of benzene rings and a side chain of benzene ring in the
RB molecule make it less adsorbable than MB. McCre-
ary and Snoeyink (1980) also found similar results. Be-
sides, RB has higher molecular weight and larger ionic
radius than MB, which further makes RB less amenable
for biosorption compared to MB. The ionic radii of MB
and RB are 15 Å and 23.3 Å, respectively (Haglund
et al., 1996), so only pores larger than 23.3 Å (ionic
radius of RB at pH 7) are accessible to RB. Several
researchers have also reported similar results (DeJohn,
1976; McCreary and Snoeyink, 1980; Carlson and
Silverstein, 1998).
3.2. Effect of ionic strength
Wastewaters from textile-manufacturing or dye-pro-
ducing industries contain various types of suspended
and dissolved compounds apart from the dyes which
can be considered as impurities in the dye removal pro-
cess. These impurities could be acids, alkalis, salts or
518 N.S. Maurya et al. / Bioresource Technology 97 (2006) 512–521

metal ions. Cations such as Na+, K+, Cu2+, Ca2+ and 70

Cr3+ are the most common metal ions present in dye 60
bearing wastewater (Correia et al., 1994). The presence
50
of ions leads to high ionic strength, which may signifi-

% Removal
NaCl-AM-RB
cantly affect the performance of the biosorption process 40
NaCl-BM-RB
(Greene et al., 1987; Ramelow et al., 1992). 30
MgCl2-AM-RB
MgCl2-BM-RB
Figs. 5–8 present the effect of ionic strength on the CaCl2-AM-RB
20 CaCl2-BM-RB
uptake of dyes. It was observed that for both biosor-
10

0
<0.00001 0.0001 0.0002 0.001 0.002 0.01 0.02 0.1 0.2
80
Ionic Strength (mol/l)

Fig. 8. Effect of ionic strength on dye uptake, biosorbent-macro

60 NaCl AM
NaCl BM
fungus based sorbent (2.8 g/L), sorbate-RB (100 mg/L), pH-7.5.
% Removal

40
bents, sorption potential decreased with increase in ionic
strength. For biosorption by AM, as ionic strength
20 changed from 0.00001 to 0.1 mol/l, the removal of MB
decreased by 19% (from 56% to 37%, Fig. 5); however,
the removal of RB decreased by 7% (from 66% to
0
0 0.1 mM 1 10 100
59%, Fig. 6). In the case of biosorption on BM, the up-
take of MB decreased by 12% (from 55% to 43%, Fig.
+
Fig. 5. Effect of Na on dye uptake, biosorbent-macro fungus based 5), while the uptake of RB decreased by 6% (from
sorbent (0.3 g/L), sorbate-MB (100 mg/L), pH-7.5.
64% to 58%, Fig. 6) as ionic strength varied from
0.00001 to 0.1 mol/l. Thus, increased ionic strength led
to a decrease in the sorption potential of biosorbents
80
NaCl AM
for dye removal. The adverse effect of ionic strength
NaCl BM on dye uptake suggests the possibility of ion exchange
60 mechanisms being in operation in the biosorption pro-
% Removal

cess. It may be due to competition of Na+ (present in

salt used to change the ionic strength of dye solution)
40 with positively charged dye molecules for the same bind-
ing sites on the biosorbent surface. Several authors
20 (Ramelow et al., 1992; Osman and Bandyopadhyay,
1996; Donmez and Aksu, 2002) have reported similar re-
sults for the sorption of heavy metals onto the biosor-
0
0 0.1 1 10 100 bent. However, in the case of sorption of RB, ionic
mM
strength had a lower impact on its uptake compared
Fig. 6. Effect of Na+ on dye uptake, biosorbent-macro fungus based to the uptake of MB. Bohrer et al. (2002) have also
sorbent (2.8 g/L), sorbate-RB (100 mg/L), pH-7.5. made similar observations. This may be due to the pres-
ence of a positive electrical charge on the chromophore
of MB. There is a carboxylic group (–COOH) in the RB
70
molecule, which imparts negative charge to the chromo-
60 phore. The positive charge is contributed by the amino
50 group. There are two amino groups in the RB molecule.
% Removal

NaCl-AM-MB
40 NaCl-BM-MB The net positive charge on the RB dye molecule is due to
MgCl2-AM-MB resonance of two RB structures as shown in Fig. 9. Be-
30 MgCl2-BM-MB
CaCl2-AM-MB
sides, the presence of a carboxylic group in the RB mol-
20 ecule adds to the negative charge of the RB molecule
CaCl2-BM-MB
10 above pH 4.2. Thus, RB in aqueous solution above
0 pH 4.2 is zwitterionic. As a result of the combination
<0.00001 0.0001 0.0002 0.001 0.002 0.01 0.02 0.1 0.2 of positive charge from the amino group, and negative
Ionic Strength (mol/l) charge from the carboxylic group, the net electrical
Fig. 7. Effect of ionic strength on dye uptake, biosorbent-macro character of the RB molecule in aqueous phase at pH
fungus based sorbent (0.3 g/L), sorbate-MB (100 mg/L), pH-7.5. 7.5 is almost neutral (Grauer et al., 1987). Therefore,
 N.S. Maurya et al. / Bioresource Technology 97 (2006) 512–521
+ +
(C2H5)2 N O N(C2H5)2
_ _
Cl
C C
COOH
Fig. 9. Resonance structure of RB.
biosorption of the RB molecule is less affected by ionic
strength in comparison to fully ionized MB (Randtke
and Jepsen, 1982).
Figs. 7 and 8 show the effect of monovalent and diva-
lent ions on the sorption potential of biosorbents. For
monovalent electrolyte, i.e., NaCl, at a lower ionic
strength, i.e. <0.001 mol/l, the uptake of both MB and
RB varied by less than 2%. For the divalent electrolytes,
i.e. CaCl2 and MgCl2, this variation was 7% for MB and
6% for RB (Figs. 7 and 8). It indicated that there was
very little variation in the dye uptake capacity of biosor-
bents for both dyes. However, at a higher ionic strength,
i.e., 0.001 to 0.2 mol/l, uptake of MB and RB varied by
13% and 9%, respectively, for monovalent ion concen-
tration. For divalent cations, when concentration of
divalent ion changed from 0.002 to 0.2 mol/l, the dye up-
take changed by 44% for MB and by 3% for RB. Thus,
at a higher molar strength (more than 0.001mol/l) the
effect of divalent ions was more prominent than that
of monovalent ions for MB. However, it was difficult
to distinguish between the effects of the divalent cations
Ca2+ and Mg2+, although it was expected theoretically
that the effect of Ca2+ would be greater than that of
Mg2+ due to its smaller hydrated radius and higher
atomic number (Helms, 1973).
3.3. Effect of pH
The pH of dye bearing wastewater varies greatly. The
interaction between sorbate and sorbent is affected by
the pH of an aqueous medium in two ways: firstly, since
dyes are complex aromatic organic compounds having
different functional groups and unsaturated bonds, they
have different ionization potentials at different pH,
resulting in the pH dependent net charge on dye mole-
cules. Secondly, the surface of the biosorbent consists
of biopolymers with many functional groups, so the
net charge on biosorbent, which could be measured in
the form of zeta potential, is also pH dependent. There-
fore, the interaction between dye molecules and biosor-
bent is basically a combined result of charges on dye
molecules and the surface of the biosorbent. The effect
of pH on the sorption of metals has been reported in de-
tail elsewhere (Schiewer and Wong, 2000; Greene et al.,
1987; Ramelow et al., 1992; Schiewer and Volesky,
1995). However, in the case of dyes, limited information
is available.
519
(C2H5)2 N O N(C2H5)2
Cl
COOH
100
AM-MB
80
AM-RB
BM-MB
% Removal
60 BM-RB
40
20
0
1 3 5 7 9 11
Initial pH
Fig. 10. Effect of initial pH on dye uptake, sorbents-BM and AM (for
MB-0.3 g/L, for RB-0.8 g/L), initial dye concentration-100 mg/L.
Fig. 10 presents the effect of pH on dye uptake using
AM and BM. In the case of MB, as pH increased from 3
to 11, uptake increased from 18% to 75% for BM, and
from 16% to 79% for AM. It is evident that in the case
of MB, uptake increased consistently with pH (Fig. 10).
This behavior of dye sorption can be explained on the
basis of surface charge of the biosorbents. It has been re-
ported that such biosorbents have net negative charge in
the aqueous phase (Kapoor et al., 1999). With increase
in pH, the net electronegativity of the biosorbent
increases due to deprotonation of different functional
groups (Schiewer and Volesky, 1995; Kapoor et al.,
1999; Harris and Ramelow, 1990). Therefore, there is
an increase in attractive electrostatic force between the
negatively charged biosorbent particle and positively
charged MB dye ion. However, uptake of RB was not
affected by the increased pH to a large extent. It de-
creased from 25% to 5% for BM, and from 22% to 9%
for AM, as pH increased from 3 to 11. In common with
MB, RB is a cationic dye, but it is an aromatic amino
acid. It possesses both acidic and basic character due
to the presence of the carboxylic group (–COOH) (Gurr,
1971). RB is an aromatic amino acid, which dissociates
in aqueous solution leaving a positive charge on one of
the nitrogens (Haglund et al., 1996). Thus, RB changes
its basicity or acidity with a change in pH, since the
pKa value for the aromatic carboxylic acid group pres-
ent on the RB molecule is 4.2 (Perrin et al., 1981). So
below pH 4.2, the RB ion contains one positive charge
on any of the nitrogens and the carboxylic group is un-
ionized (Table 2). At a higher pH, i.e. above 4.2, there is
an increase in the net negative charge on the biosorbent
520 N.S. Maurya et al. / Bioresource Technology 97 (2006) 512–521
surface, and the carboxylic group ionizes, leaving a neg-
ative charge on the RB molecule. Net positive charge on
RB gets reduced. Thus, at a higher pH, a significantly
lower electrostatic attraction exists between almost neu-
tral RB and negatively charged biosorbent, so less sorp-
tion of RB is expected. Both biosorbents showed the
same pattern (increasing for MB and decreasing for
RB) for both the dyes as pH increased. Uptake capacity
of MB increased and that of RB decreased as the initial
pH of the dye solution increased. Previous researchers
have reported similar observations for the sorption of
MB (Gupta et al., 2000; Ramakrishna and Viraragha-
van, 1996; El-Nabarwy and Khedr, 2000; Kamel,
1993) and RB (Gupta et al., 2000; Namasivayam
et al., 1996).
Since the dye solution was not buffered, a change in
pH of the dye solution is expected during the sorption
process. So, both initial and equilibrium pH were
measured. It was observed that the final pH was lower
than the initial pH for the biosorption of MB onto both
of the biosorbents. This indicated release of H+ ions
during the biosorption of MB. The release of H+ ions
may be indicative of an ion exchange type of sorption
mechanism. However, in the case of RB, at initial pH
values between 3 and 5, final pH was higher than the
initial pH, indicating the possibility of either release
of OH
ions into solution or the uptake of H+ ions
from the solution.
4. Conclusions
In the present study, macro fungus-based biosorbents
were applied successfully for the sorption of cationic
dyes, namely Methylene Blue and Rhodamine B. Equi-
librium sorption data were modeled using the Langmuir
and Freundlich equations. Although experimental equi-
librium data obtained for all sorbent-sorbate systems
were well described by both models, the Langmuir equa-
tion provided a better fit than the Freundlich. For MB,
monolayer sorption capacities were 202.38 mg/g and
232.73 mg/g and for RB, 36.82 mg/g and 25.12 mg/g,
using biosorbents AM and BM respectively. MB was
more adsorbable than RB. Higher molecular weight, lar-
ger ionic size and presence of carboxylic group were the
probable reasons for the lower sorption of RB. Sorption
of MB was dependent on initial pH of solution and the
presence of cations. The presence of cations and in-
creased ionic strength had adverse effects on dye sorp-
tion. Sorption of MB was more affected by the ionic
strength than RB. The presence of a carboxylic group
and its dissociation behavior was probably the reason
why RBs sorption was less affected by pH and increased
ionic strength. Sorption of MB increased whereas sorp-
tion of RB decreased with an increase in pH from 3 to
11.
Acknowledgements
The authors are grateful to the German Academic
Exchange Service (DAAD) for providing financial sup-
port to Mr. N.S. Maurya to visit the Institute WAR,
Technische Universität Darmstadt, Germany to carry
out part of this work.
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