Environ Monit Assess (2012) 184:66076611

DOI 10.1007/s10661-011-2445-4

Analysis of trace elements (Cu, Cd, and Zn) in the muscle,

gill, and liver tissues of some fish species using anodic
stripping voltammetry
S. Sobhanardakani & L. Tayebi & A. Farmany &
M. Cheraghi

Received: 11 September 2011 / Accepted: 2 November 2011 / Published online: 19 November 2011
# Springer Science+Business Media B.V. 2011

Abstract Heavy metal (Cu, Cd, Zn) concentration of
the muscle, gill, and liver of Otolithes rubber,
Pampus argenteus, Parastromateus niger, Scomber-
omorus commerson, and Onchorynchus mykiss are
determined by anodic stripping voltammetry method.
The results show that the concentration of Cu, Cd,
and Zn varied between 0.44 and not detected, 0.585
and 0.001, and 0.450 and 0.005 g/g, respectively. It
is interesting that in O. mykiss, the mean value of
copper concentration was more than other marine
organisms of this study, while in P. niger, the mean
value of copper was minimum. The results of this
study indicate that in the different tissues of fish
species (O. rubber, P. argenteus, P. niger, S. commer-
son, and O. mykiss), the concentrations of Cu, Cd, and
S. Sobhanardakani (*) : M. Cheraghi
Department of Environment, Hamedan Branch,
Islamic Azad University,
Hamedan, Iran
e-mail: s_sobhan@iauh.ac.ir
L. Tayebi
Department of Environment,
Malayer University,
Malayer, Iran
A. Farmany
Young Researchers Club, Hamedan Branch,
Islamic Azad University,
Hamedan, Iran
e-mail: a.farmany@usa.com
Zn were significantly lower than the permissible
FAO/WHO levels.
Keywords Heavy metal . Fish tissues . Otolithes
rubber . Pampus argenteus . Parastromateus niger .
Scomberomorus commerson . Onchorynchus mykiss
Introduction
Today, one of the most serious problems facing the
world is contamination of the environment by
inorganic, organic, and organometallic materials.
There has been growing interest in the monitoring of
heavy metals in the bio-organs (Kalay et al. 1999;
Rose et al. 1999). Fish tissues may contain heavy
metals from their presence in the water or fish foods
(McLaughlin et al. 1999). The accumulation of metals
varies greatly between both fish species and/or fish
tissues. Generally, fish could translocate the larger
quantities of heavy metals in the gill, liver, and
muscle tissues (Kagi and Schaffer 1998). For the
normal metabolism of fishes, essential metals such as
copper must be taken up from water, food, or
sediment (Canli and Atli 2003). Cadmium level can
be particularly elevated in the kidneys of marine
mammals (Dietz et al. 1998). Its potential toxic effects
are mitigated by binding to the metallothioneins both
in the liver and kidney tissues (Klaassen and Liu
1997; Teigen et al. 1999). Also, Cd could be stored
over a very long time in the renal tissue of marine
6608
mammals. Zn is included in the list of elements in which
their physiological role and necessity for plant life is
proved. Recently, analysis of heavy metals in the different
tissues of fish has been broadly investigated (Emami
Khansari et al. 2005; Fernandes et al. 2008; Voegborlo
et al. 1999; Al-Kahtani 2009; Suppin et al. 2005;
Taghipour and Azizi 2011; Blasco et al. 1999; Bird et
al. 1998; Alabaster and Lloyd 1994; Deshpande et al.
2009). The present study reports the heavy metal
(Cu, Cd, and Zn) analysis of different tissues (muscle,
gill, and liver) of fish species (Otolithes rubber,
Pampus argenteus, Parastromateus niger, Scomber-
omorus commerson, and Onchorynchus mykiss) by
anodic stripping voltammetry method.
Experimental
Chemical and reagents
All chemical reagents were of analytical reagent grade,
purchased from Merck (Germany). All solutions were
prepared with doubly distilled water. Stock standard
solution of Cu(II), Cd(II), and Zn(II) (1,000 mg/l) were
prepared by dissolving the appropriate amount of metal
salts in doubly distilled water and diluting to 1,000 ml in
the volumetric flask. As supporting electrolyte, 0.1 M
acetateacetic acid buffer (pH 4.5) was used.
Apparatus
All voltammetric measurements were carried out
using a polarographic processor, model 746 VA
(Metrohm), in combination with a polarographic
stand model 747 VA (Metrohm). The electrode stand
consists of a hanging mercury drop electrode
(HMDE) as working electrode, a double junction
Ag/AgCl (3 M KCl, saturated AgCl, and 3 M KCl in
the bridge) as reference electrode, and platinum wire,
with considerably larger surface area than that of
HMDE, as auxiliary electrode. All potentials quoted
are relative to the Ag/AgCl reference electrode.
Stirring was carried out by a large Teflon road with
2,000 rpm speed. A 780 pH Meter (Metrohm),
equipped with a combined Ag/AgCl glass electrode
was used for pH measurements. Eppendorf reference
variable micropipettes were used to pipette microliter
volumes of solutions. All glasswares were soaked
overnight in 10% (v/v) nitric acid, followed by
Environ Monit Assess (2012) 184:66076611
washing with 10% (v/v) hydrochloric acid, and rinsed
with doubly distilled water and dried before using.
Sample preparation
Although voltammetric techniques are inherently
precise and accurate, the results obtained using these
techniques may be invalidated due to contamination
caused by poor sample handling and preparation.
Therefore, stringent conditions should be routinely
used for trace analysis. For example, all reagents,
standard solutions, etc. should be ultrapure, and all
glasswares need to be scrupulously cleaned. Similarly,
stringent conditions should also be used for sampling
and the pretreatment of samples; these two stages
should be simplified as much as possible to minimize
the potential for sample loss and contamination.
Different complex matrices of the analytical samples
require the prior mineralization for most analytical
methods. This step is critical in the analytical
procedure for the determination of metal concentra-
tion. Sample digestion techniques, such as microwave
and conventional acid digestion method, have been
used widely for the dissolution of target elemental
analysis. These digestion techniques require the use of
concentrated mineral acids and high temperatures
(Jalbani et al. 2007). In this study fish samples were
cleaned with distilled water and then dissected. Two
grams of muscle, liver, or gill tissue of fish samples
was removed and weighed for the analysis. For
estimation of heavy metal content, 2 g of each tissue
was taken in a 100-ml Borosil beaker. To this, 2 ml of
HNO3 and 1 ml of HClO4 were added and kept for
digestion on a hot plate at 100C till complete
digestion was achieved. It was ensured that the
residue obtained after digestion was free from organic
matter which acts as impurities in metal analysis
(Khandekar et al. 1984; Raghunath et al. 1997;
Deshpande et al. 2009).
Table 1 Determination of Cu(II), Cd(II), and Zn(II) in the
muscle, gill, and liver of O. rubber (n=3)
Metal ion Muscle (g/g) Gill (g/g) Liver (g/g)
Cu(II) 0.007 0.010
Cd(II) 0.004 0.086 0.008
Zn(II) 0.4 0.05 0.15
Environ Monit Assess (2012) 184:66076611 6609

Table 2 Determination of Cu(II), Cd(II), and Zn(II) in the Table 4 Determination of Cu(II), Cd(II), and Zn(II) in the
muscle, gill, and liver of P. argenteus (n=3) muscle, gill, and liver of S. commerson (n=3)

Metal ion Muscle (g/g) Gill (g/g) Liver (g/g) Metal ion Muscle (g/g) Gill (g/g) Liver (g/g)

Cu(II) 0.031 0.19 0.047 Cu(II) 0.016 0.021 0.057

Cd(II) 0.006 0.004 0.005 Cd(II) 0.007 0.004 0.004
Zn(II) 0.25 0.03 0.033 Zn(II) 0.005 0.062 0.013

Sample analysis analysis. In an analysis of heavy metals by anodic

To the analysis of Zn(II), Cu(II), and Cd(II) concen-
tration in the muscle, gill, and liver of fish species (O.
rubber, P. argenteus, P. niger, S. commerson, O.
mykiss), 5 ml of each sample solution and 1 ml
acetateacetic acid buffer solution were transferred
into the electrochemical cell and diluted to 10 ml by
doubly distilled water. The solution was deaerated by
passing pure nitrogen for 5 min. The deposition
potential was controlled at (0.75, 1.0, and 0.25
for Cd, Zn, and Cu, respectively) and applied to a
fresh mercury drop while the solution was stirred.
After the deposition step and further 10 s (equilibrium
time), the voltammograms were recorded. Different
concentrations of the standard metal ions were added
to the cell. The solution was stirred and purged with
nitrogen for 1 min after each spike. Finally, the
concentrations of Cd(II), Zn(II,) and Cu(II) were
calculated in the sample solutions by using the
standard addition method.
Results and discussions
Anodic stripping voltammetry is a powerful electro-
chemical technique, namely due to its high sensitivity,
repeatability, low detection limit, low cost of equip-
ment, as well as to its possibility of performing in situ
field analysis. An important advantage of this tech-
nique lies in its ability to trace and ultratrace level
stripping voltammetry, the procedure is essentially
involving two steps: the predeposition step which
consists the reducing of metal ions onto the surface of
working electrode by means of applying an adequate
cathodic potential in a certain period. In this step, the
mass transport may be enhanced by means of stirring
road. In the final step, the metal ions that are
deposited in the previous step are reoxidized back
into the solution by means of applying an anodic
potential sweep. The voltammograms recorded in this
step (currentpotential plot) provide the analytical
signals. Because of high sensitivity of anodic strip-
ping voltammetry, this method is applied to the
determination of Cu, Cd, and Zn in the different
tissues (muscle, gill, and liver) of fish species (O.
rubber, P. argenteus, P. niger, S. commerson, and O.
mykiss). The results of the heavy metal analysis of the
muscle, gill, and liver of O. rubber are presented in
Table 1. As shown in Table 1, no copper is detected in
the liver tissue of the O. rubber. Zn level in muscle
tissue of O. rubber was higher than gill and liver
tissue of this marine organism. Also, in this marine
organism, the maximum concentration of Cd was
found in the gill tissue. The results of heavy metal
analysis of the P. argenteus are presented in Table 2.
The maximum concentration of Cu, Cd, and Zn was
found in the gill tissue of this marine organism. The
results of Cu(II), Zn(II), and Cd(II) analysis of P.
niger are presented in Table 3. As shown in Table 3,
no copper ion is detected in the muscle and gill tissues

Table 3 Determination of Cu(II), Cd(II), and Zn(II) in the Table 5 Determination of Cu(II), Cd(II), and Zn(II) in the
muscle, gill, and liver of P. niger (n=3) muscle, gill, and liver of O. mykiss (n=3)

Metal ion Muscle (g/g) Gill (g/g) Liver (g/g) Metal ion Muscle (g/g) Gill (g/g) Liver (g/g)

Cu(II) 0.009 Cu(II) 0.23 0.34 0.44

Cd(II) 0.45 0.09 0.585 Cd(II) 0.001 0.004 0.002
Zn(II) 0.011 0.047 0.45 Zn(II) 0.029 0.034 0.012
6610
of P. niger. The dispersion of Cd, Zn, and Cu level in
P. niger shows that the maximum concentration of
these heavy metals is in the liver tissue of this marine
organism. The results of heavy metal analysis of the
muscle, gill, and liver tissues of S. commerson are
presented in Table 4. As shown in Table 4, in S.
commerson, the maximum copper concentration is in
the liver tissue. Also, Cd and Zn content of the
muscle and gill tissues of this marine organism is
more than the other tissues. Heavy metal analysis of
the gill, muscle, and liver tissues of O. mykiss is
presented in Table 5. As shown in Table 5, in order to
the other marine organisms studied here, Cu level in
the muscle, gill, and liver tissue of O. mykiss was
maximum. Also, in different tissues of this marine
organism, the Cd content is low. The results of this
study shows that the copper concentration reaches to
maximum in the liver of O. mykiss (0.44 g/g) and
followed by the liver of O. rubber and the muscle and
gill of P. niger (not detected). Cadmium content
reaches to maximum in the liver of P. niger
(0.585 g/g) and followed by the muscle of O. mykiss
(0.001 g/g). Zinc concentration is highest in the liver
of P. niger (0.45 g/g) and followed by the muscle of
S. commerson (0.005 g/g). The results show that the
concentration of Cu, Cd, and Zn varied between 0.44
and not detected, 0.585 and 0.001, and 0.45 and
0.005 g/g, respectively. Cu, Cd, and Zn levels
obtained in this study are in agreement with previous
results reported in the literature (Canli and Atli 2003;
Emami Khansari et al. 2005; Fernandes et al. 2008;
Voegborlo et al. 1999; Al-Kahtani 2009; Suppin et al.
2005; Taghipour and Azizi 2011; Blasco et al. 1999;
Bird et al. 1998; Deshpande et al. 2009). The results
of this study indicate that the concentration of Cu, Cd,
and Zn is significantly lower than the permissible
level (FAO/WHO 1972).
Conclusion
In this paper, anodic stripping voltammetry method was
applied to the determination of Cu (II), Cd(II), and Zn
(II) in the different tissues (muscle, gill, and liver) of O.
rubber, P. argenteus, P. niger, S. commerson, and O.
mykiss. It was found that copper concentration
reaches to maximum in the liver of O. mykiss
(0.44 g/g) and followed by the liver of O. rubber
and the muscle and gill of P. niger (not detected).
Environ Monit Assess (2012) 184:66076611
Cadmium content reaches to maximum in the liver of
P. niger (0.585 g/g) and followed by the muscle of
O. mykiss (0.001 g/g). Zinc concentration is highest
in the liver of P. niger (0.45 g/g) and followed by
the muscle of S. commerson (0.005 g/g).The results
of heavy metal analysis indicate that heavy metal
content of the analyzed fish species is well below the
permissible levels.
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