Pak. J. Bot., 43(6): 2997-3000, 2011. REMOVAL OF Pb(II), Cu(II) AND Cd(II) FROM AQUEOUS SOLUTION BY SOME FUNGI AND NATURAL ADSORBENTS IN SINGLE AND MULTIPLE METAL SYSTEMS AMNA SHOAIB

Pak. J. Bot., 43(6): 2997-3000, 2011. 
REMOVAL OF Pb(II), Cu(II) AND Cd(II) FROM AQUEOUS SOLUTION BY SOME 
FUNGI AND NATURAL ADSORBENTS IN SINGLE AND MULTIPLE METAL SYSTEMS 
AMNA SHOAIB
*
, TASKEEN BADAR AND NABILA ASLAM 
Institute of Agricultural Sciences, University of the Punjab, Quaid-e-Azam Campus, Lahore-54590, Pakistan. 
*Corresponding author E-mail: aamnaa29@yahoo.com 
Abstract
Six fungal and 10 natural biosorbents were analyzed for their Cu(II), Cd(II) and Pb(II) uptake capacity from single, 
binary and ternary metal ion system. Preliminary screening biosorption of assays revealed 2 fungi (Aspergillus niger and 
Cunninghamella echinulata) and three natural [Cicer arietinum husk, Moringa oleifera flower & soil (clay)] adsorbents hold 
considerable high adsorption efficiency and capacity for 3 metal ions amongst the adsorbents. Further biosorption trials with 
five elected adsorbents showed a considerable reduction in metal uptake capability of adsorbents in binary- and ternary 
systems as compared to singly metal system. Cd(II) manifested the highest inhibitory effect on the biosorption of other metal 
ions, followed by Pb(II) and Cu(II). On account of metal preference, the selectivity order for metal ion towards the studied 
biomass matrices was Pb(II) (40-90%) > Cd(II) (2-53%) > Cu(II) (2-30%). 
Introduction 
Biosorption is a practice that utilizes inexpensive 
biomass to sequester toxic heavy metals and is 
particularly useful for the removal of trace levels of 
contaminants from industrial effluents (Volesky, 2003). 
Since 1970s, after discovery of biosorption technique, 
scientists from all over the world have been engaged in 
identifying promising biosorbent owing to availability 
several kinds of adsorbents including microorganisms i.e., 
fungi (Zhang, 2009), bacteria (Hamzah et al., 2009), algae 
(Qunaibit et al., 2005) and plants (Melcakova and 
Ruzovic, 2010). After few decades, scientists have 
diverted their attentions to explore some low cost, 
naturally abundant and environment friendly adsorbents. 
In this case they have noticed tremendous metal 
sequestering ability of various type of agro waste (Qaiser 
et al., 2009). It is however imperative to notice, that all 
studies on biosorption have provided insight into the 
identification of several microbial biomass types for 
single-metal-ion solutions. The fact cannot be ignored that 
single toxic metallic species rarely exist in wastewaters. 
Infact, the presence of multiple metal ions often causes an 
interactive effect while insufficient attention seems to 
have been given to this problem (Choy  et al., 2000, 
Hammaini et al., 2002, Sheng et al., 2007).  
Relatively few studies on multimetal systems have 
been reported, though multimetal competitive interactions 
in solution with the sorbent material are amongst the basic 
factors affecting the degree of metal removal by 
biosorption (Ceribasi & Yetis, 2001, Javaid, 2008). For 
instance, Yan & Viraraghavan (2001) explored the 
biosorption ability of  Mucor rouxii for Pb(II), Cd(II), 
Ni(II) and Zn(II) in binary and multi-metal ion solution. 
Mahvi et al., (2005) recorded 60-90% removal for Pb(II), 
Ni(II) and Cd(II) through application of tea waste from 
single metal solution and noticed a 3.5% decrease for 
Pb(II) adsorption whereas 12.2% for Ni(II) in mixture 
form Akar et al., (2006) explored the greater biosorption 
potential (22.79 mg g
-1
) of Ganoderma carnosum for the 
removal of Pb(II) ion from aqueous solution and industrial 
effluents. In a similar investigation, the biosorption of 8 
different metal ions i.e.,  Fe(II), Ni(II), Mn(II), Cu(II), 
Cd(II), Cr(III), Pb(II) and Zn(II) from aqueous solutions 
of combined industrial effluent by  Phanerochaete 
chrysosporium was investigated with following affinity 
order: Cd(II) >Zn(II)> Mn(II)> Fe(II)> Ni(II)> Cr(III)> 
Cu(II)> Pb(II) by the test fungus (Pogaku & Kulkarni, 
2006). Javaid, (2008) in her experimentation found 
potential capability of  Pleurotus ostreatus,  Ganoderma 
lucidum and Schizophyllum commune in adsorbing Cu(II), 
Ni(II), Cr(VI) and Zn(II) in single, multiple and real 
industrial solutions. However, like most of previous 
findings she noticed considerable reduction in sorption 
capacity of three fungi in competitive form and due to 
change in ionic strength rather than competition between 
the heavy metals. The present study aims to investigate 
the biosorption of divalent metal ions viz., Pb, Cd and Cu 
from single, binary and multi-component aqueous 
solutions using biomass of some fungi and plants.  
Materials and Methods 
Preparation of adsorbents:  Six fungal and  10 natural 
adsorbents were selected for biosorption trials. The pure 
cultures of six fungal species viz.,  Aspergillus niger
(FCBP 0074), Aspergillus terreus (FCBP 0058), Rhizopus 
arrhizus (FCBP 800),  Fusarium  sp., (FCBP 734), 
Trichoderma harzianum (FCBP 0139) and 
Cunninghamella echinulata (FCBP 0104) were provided 
by First Fungal Culture Bank (FCBP), in the Institute of 
Plant Pathology (IPP). A medium with 2 g L
-1 
malt extract 
was used for the cultivation  of fungi  and biomass 
preparation (Javaid & Bajwa, 2008). 
Amongst natural,  Azadirachta indica leaves,  Pinus 
sp. bark and  Moringa oleifera flower and leaves were 
collected from Botanical garden of Punjab University. 
However, Citrus reticula peels, Oryza sativa straw, Luffa 
cylindrica dried fruit and  Cicer arietinum husk was 
acquired from local market. All adsorbents were washed 
thoroughly with tap water to remove dust and twice with 
distilled water. Finally, these candidate biosorbents were 
dried in oven at 100
o 
C for 24 hours and homogenized in a 
blender to break the cell aggregates into smaller fragments 
of 0.5-1 mm diameter (mesh size 150  μm). Waste 
charcoal was taken from Natural Product laboratory of 
Herbal Heritage Centre, IPP, Punjab University, and soil 
(clay) was collected from IPP lawn. Both of the 
biosorbents were dried at 60°C in an oven for 24 hours 
and sieved (mesh size 150μm) . The dried biomass of each 
adsorbent was utilized in biosorption experiments. 
Batch screening experiments: The stock metal solutions 
at various concentrations were prepared by using nitrate 2998  AMNA SHOAIB ET AL., 
salts of Pb(II), Cu(II) and Cd(II) (Merck, Germany). 
Preliminary screening batch experiments were conducted 
with 6 fungal and 10 natural biosorbents for each of three 
metal ions.  Metal biosorption experiments were carried out 
in a 250mL flask at 25 ± 1°C in a rotary shaker at 150rpm. 
The flask was filled with 100mL (1000mg L
-1
) of 
previously prepared solutions of each metal. An arbitrary 
amount of biosorbent biomass (0.1 g of dry cell weight mL
-
1
) was used. Each experiment was conducted for 4 hours, 
which was enough time to achieve steady state biosorption. 
The pH of each reaction mixture in flask was adjusted to 
the 4.5 using 0.1 N HNO3 or 0.1 N NaOH. After desired 
contact time, the mixture was filtered through Whatmann 
filter paper No.1 and filtrate  was analyzed using atomic 
absorption spectrophotometer (BMS-100 SERIES) to find 
out the amount of metal left after sorption. 
Batch Experiments for binary- and multiple metal 
systems: Potential adsorbents obtained through biosorption 
screening trials were further subjected to batch 
examinations in binary and multiple metal mixture. For this 
purpose each metal ions [Cu(II), Cd(II) and Pb(II)] was 
taken in the equivalent concentration within the range of 
100 mg L
-1
. The proposed binary mixtures were in 
following combinations: Cu-Cd, Cu-Pb & Cd-Pb, whereas 
a grouping of Cu-Cd-Pb was taken as ternary aqueous 
phase. The adsorption experiment was carried out in a 
similar fashion as was performed for single metal cases. 
Biosorption data evaluation: The amount of metallic ion 
biosorbed per gram of biomass (q) and the efficiency of 
biosorption (E) were calculated using following 
equations: 
q = 
⎝
⎛
⎠
Ci
⎞
-Cf
m
 V     and    E = 
⎝
⎛
⎠
Ci
⎞
-Cf
Ci
 x 100  
where, Ci
 = initial concentration of the metallic ion (mg L
-
1
); Cf
 = final concentration of metallic ion (mg L
-1
); m = 
dried mass of the biosorbent in the reaction mixture (g) 
and V = volume of reaction mixture (mL). 
       
Results and Discussion 
Quantitative screening of efficient biosorbents:  Data 
presented in Table 1 reveals residual metal ion 
concentration (Cf) and biosorption efficiency (E) of 
different biosorbents with respect to their metal 
sequestering potential. Results presented clearly indicates 
that all selected adsorbents exhibited the highest removal 
efficiency for Pb(II) (40-90 %) in comparison to Cd(II) 
(2-53 %) and Cu(II) (2-30 %). In case of Cu(II) trend of 
biosorption by various adsorbents was recorded to follow 
the sequence of:  M. oliefera flowers (50%) >  C. 
echinulata and soil (30%) >  A. niger and  C. arietinum
(21%). Assorted adsorbents possess following 
predilection for Cd(II) ions: M. oliefera flowers (53%) > 
C. echinulta, soil(clay)  Citrus reticula peels, M. oliefera
and  A. indica leaves (50%) >  A. niger and  A. terreus
(43%). For Pb(II) ions different metal sequesters tag along 
the following selectivity order : six fungal adsorbents (80-
90%) >  M. oliefera (leaves & flowers),Citrus reticula
peels, Pinus bark and soil (80%) > O. sativa husk (60%). 
Thus, screening experiments demonstrated that two fungal 
species viz.  A. niger and  C. echinulta and three natural 
adsorbents i.e. M. oliefera flowers, C. arietinum husk and 
soil (clay) hold considerable greater biosorption efficiency 
and capacity for three metal ions (Cu, Cd & Pb) in 
comparison to rest of adsorbents. Therefore, these five 
efficient adsorbents were chosen for further biosorption 
assays. The assessment regarding screening of potent 
adsorbents for Cu(II), Cd(II) and Pb(II) revealed variable 
biosorption capacity of test species. Disparity in 
biosorption capacity of different adsorbents may be 
ascribed to the intrinsic ability of organism, its chemical 
composition of cell wall leading various types of 
interaction of metals with adsorbents (Gadd, 1993).  
Table 1. Comparative representation of biosorption efficiency (%) of various biosorbents for metal ions. 
Biosorption conditions: biosorbents concentration, 0.1g 100 mL
-1
; pH, 4.5; 150 rpm at 25
°
C for 4 hours. 
Pb(II) Cu(II) Cd(II) 
Biosorbents Co g L
-1
Ce g L
-1
 E % Ce g L
-1
 E % Ce g L
-1
 E % 
1.  Aspergillus  niger  100 10 90 79 21 57 43 
2.  Rhizopus arrhizus  100 13 87 92 8 82 18 
3.  Cunninghamella echinulata  100 15 85 70 30 50 50 
4.  Fusarium sp.  100 17 83 90 10 60 40 
5.  Trichoderma harzianum  100 18 82 98 2 85 15 
6.  Aspergillus terreus  100 19 81 90 10 57 43 
7.  Citrus reticula peels 100 20 80 92 8 50 50 
8. Pinus sp. bark 100 20 80 90 10 98 2 
9.  Moringa oleifera flowers 100 20 80 50 50 50 53 
10.  Moringa oleifera leaves 100 20 80 90 10 50 50 
11. Soil (clay) 100 20 80 70 30 50 50 
12.  Oryza sativa husk 100 40 60 96 4 70 30 
13.  Azadirachta indica  leaves 100 60 40 84 16 52 48 
14.  Luffa cylindrica dried fruit 100 60 40 84 16 90 10 
15.  Cicer arietinum husk 100 60 40 79 21 60 40 
16. Charcoal 100 60 40 88 12 90 10 REMOVAL OF Pb, Cu AND Cd) FROM AQUEOUS SOLUTION  2999
Biosorption assays in binary and multiple metal 
systems: A comparison of the effect of Cu(II), Cd(II) and 
Pb(II) adsorption by selected adsorbents viz. A. niger, C. 
echinulta, M. oliefera flowers, C. arietinum husk and soil 
(clay) in single-, binary- and ternary metal systems is 
pooled in Tables 2 & 3. Data acquired revealed that, all 
five adsorbent species exhibited considerable metal 
uptake potency in the binary and ternary mixture. 
However, a considerable reduction in metal sequestering 
ability of the adsorbents was evident in binary and 
multiple metal systems in comparison to single metal 
system. Thus, in case of binary metal system all the 
chosen adsorbents exhibited the highest decline of 5-20% 
in adsorption efficiency for Cu(II) followed by 2-18%for 
Pb(II) and 5-12% for Cd(II) in comparison to single metal 
system  (Table 2). While, a net reduction of 18-25%, 17-
25% and 18-21% in uptake efficiency of adsorbents for 
Cu(II), Pb(II) and Cd(II), respectively was traced in 
ternary metal system in contrast to metal confiscating 
potential in single metal system (Table 3). The results in 
the binary and ternary systems clearly showed that the 
combined action of multiple ions was antagonistic. Thus, 
the metal removal efficiency was greater in the singlecomponent systems in the comparison with the multicomponent one. It is probably due to the absence of 
competitive processes between metals and biomass in 
single component system (Kovacevic  et al., 2000). The 
most likely reason for the antagonistic effect is the 
competition for adsorption sites on the cell surfaces and/or 
the screening effect by the competing metal ions (Sheng et 
al., 2007). Results of present research also showed that 
Cd(II) exerted the most inhibitory effect on the 
biosorption of other metals, followed by Pb(II) and Cu(II). 
A similar phenomenon had been observed in the binary 
adsorption of Pb(II), Cu(II), Cd(II), and Ni(II) with a 
natural heterogeneous sorbent, where it was shown that 
Cd(II) and Ni(II) strongly competed with each other and 
were displaced in the presence of Pb(II) and Cu(II) 
(Papini et al., 2004). 
Table 2. Comparative biosorption efficiency of various adsorbents in single metal systems (SMS) 
and binary metal systems (BMS) at 100 mg L
-1
. 
Cu(II) Cd(II) 
 Biosorbents 
SMS BMS SMS BMS 
1.  Aspergillus niger  21% 18% 43% 41% 
2.  Cunninghamella echinulata  30% 25% 50% 48% 
3.  Cicer arietinum husk 21% 19% 40% 38% 
4.  Moringa oliefera flowers 50% 44% 53% 50% 
5. Soil (clay) 30% 24% 50% 48% 
Cu(II) Pb(II) 
 Biosorbents 
SMS BMS SMS BMS 
1.  Aspergillus niger  21% 20% 90% 76% 
2.  Cunninghamella echinulata  30% 24% 85% 77% 
3.  Cicer arietinum husk 21% 20% 40% 37% 
4.  Moringa oliefera flowers 50% 45% 80% 78% 
5. Soil(clay) 30% 25% 80% 70% 
Cd(II) Pb(II) 
 Biosorbents 
SMS BMS SMS BMS 
1.  Aspergillus niger  43% 40% 90% 75% 
2.  Cunninghamella echinulata  50% 45% 85% 70% 
3.  Cicer arietinum husk 40% 38% 40% 33% 
4.  Moringa oliefera flowers 53% 47% 80% 70% 
5. Soil (clay) 50% 44% 80% 68% 
  
Table 3. Comparative biosorption efficiency of various adsorbents in single metal systems (SMS) and ternary 
metal systems (TMS) at 100 mg L
-1
. 
Cu(II) Cd(II) Pb(II) 
 Biosorbents 
SMS TMS SMS TMS SMS TMS 
1.  Aspergillus niger  21% 16% 43% 35% 90% 67% 
2.  Cunninghamella echinulata  30% 22.5% 50% 41% 85% 63% 
3.  Cicer arietinum husk 21% 16% 40% 32% 40% 30% 
4.  Moringa oliefera flowers 50% 41% 53% 42% 80% 66% 
5. Soil (clay) 30% 23% 50% 40% 80% 61% 
Among the three metal ions, the five adsorbents 
exhibited the highest adsorption efficiency for Pb(II) and 
lowest for Cu(II). Cd(II) manifested the highest inhibitory 
effect on the biosorption of other metal ions, followed by 
Pb(II) and Cu(II). Generally, all the adsorbent 
demonstrated the highest uptake efficacy for Pb(II), 
followed by Cd(II) and Cu(II). Sheng  et al., (2007) 
observed following preference order of metal ions: Pb(II) 
> Cu(II) > Cd(II) onto algal biomass and correlate this 
uptake trend with the electronegativities of the metal-ion 
hydroxides. The electronegative values of Pb(II) is 2.33, 
Cu(II): 1.90, Cd(II): and 1.69. According to them, higher 3000  AMNA SHOAIB ET AL., 
electronegativity corresponds to a higher adsorption 
capacity owing to higher attraction of metal ions for 
electrons (Wang  et al., 2006). In current study, the 
domino effect of Cd(II) over Cu(II), may be due to 
different pH value employed during biosorption 
experiments and dissimilarity in the nature of biomass. 
Ionic radius and hydration energy is an important factor in 
sorption process. The preferential sorption behavior of 
adsorbents for metal ions acquired in present 
investigations could also be explained in terms of ionic 
radii of the metal ions (Cu= 0.73 Å; Cd = 0.97 Å; Pb = 
1.19 Å). Thus, the element with larger ionic radius will 
compete faster for exchange sites than those of smaller 
ionic radius. Adsorption may be related to the loss of the 
entire hydration sphere that precedes hydrolysis. 
According to Horsefall & Spiff (2005), smaller the ionic 
radius, the greater its tendency to hydrolyze leading to 
reduce sorption. The observed order indicates that Pb(II) 
may have greater accessibility to the surface of certain 
pores than Cd(II) and Cu(II) due to its larger ionic radius. 
Results obtained from the present study conclude that: 
1. Cunninghamella echinulata and  Moringa oleifera
flowers possess substantial adsorption potential for 
Cu(II), Cd(II) and Pb(II) in single- binary and ternary 
metal solution at metal concentration 100 mg L
-1
.  
2. The sorption of metal ions was reduced by the presence 
of co-ion(s), with the inhibitory effect increasing as the 
concentration of the co-ion(s) increased.  
3. Adsorbents hold maximum uptake efficiency and 
capacity for Pb(II), followed by Cd(II) and Cu(II). 
4. Cd(II) manifested the highest inhibitory effect on the 
biosorption of other metal ions, followed by Pb(II) 
and Cu(II). 
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(Received for publication 28 November 2010)