Food Chemistry 124 (2011) 1005–1010

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Food Chemistry
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Metal content determination in four fish species from the Adriatic Sea
Nina Bilandžić *, Maja Ðokić, Marija Sedak
Department of Veterinary Public Health, Laboratory for Residue Control, Croatian Veterinary Institute, Savska cesta 143, HR-10000 Zagreb, Croatia

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

Article history: Concentrations of As, Cd, Cu, Hg and Pb were measured by atomic absorption spectrometry (AAS) in mus-
Received 19 February 2010 cle tissues of four fish species: anchovy (Engraulis encrasicholus), mackerel (Scomber japonicus), red mullet
Received in revised form 9 July 2010 (Mullus surmuletus) and picarel (Spicara smaris) from the Croatian waters of the Adriatic Sea during 2008
Accepted 19 July 2010
and 2009. Metal levels measured in anchovy were in the following ranges (mg kg 1): As 0.01–54.8, Cd
0.001–0.02, Cu 0.001–6.29, Hg 0.001–0.52 and Pb 0.001–0.34 mg kg 1. Metal ranges in red mullet were
(mg kg 1): As 0.01–70.9, Cd 0.002–0.85, Cu 0.001–57.3, Hg 0.001–2.07 and Pb 0.001–0.27 mg kg 1. Metal
Keywords:
level ranges measured in mackerel were (mg kg 1): As 0.01–36.4, Cd 0.001–0.1, Cu 0.001–15.9, Hg
Metal levels
Four fish species
0.001–0.78 and Pb 0.002–0.24 mg kg 1. In picarel, metal level ranges were (mg kg 1): As 0.01–54.6, Cd
Adriatic Sea 0.001–0.1, Cu 0.08–32.9, Hg 0.001–0.207 and Pb 0.001–0.46 mg kg 1. Significant differences in metal
Croatia concentrations were found among fish species. The results presented on metal contents in the examined
species give an indication of the environmental conditions. Concentrations of Cd, Cu, Hg and Pb obtained
were far below the established values by the European Community regulations. However, arsenic levels
found in red mullet were higher than the recommended legal limits for human consumption and as such
may present a human health issue.
Ó 2010 Elsevier Ltd. All rights reserved.

1. Introduction
Heavy metal toxins have been present in every segment of hu-
man life for an immeasurable amount of time due to our increased
use of such compounds. Heavy metal toxins can induce a variety of
adverse health effects in an exposed individual, such as behav-
ioural, physiological, and cognitive effects, depending on the type
of toxin. Heavy metals are discharged into the marine environment
through urban discharge, agriculture, mining, combustion and
industrial discharge, and can remain in solution or in suspension
and precipitate to the bottom, or be taken up by organisms, thus
creating a potential source of heavy metal pollution in the aquatic
environment. Accordingly, metals can accumulate to toxic concen-
trations and cause ecological damage, and subsequently be trans-
ferred to humans through the food chain (Giordano et al., 1991;
Gümgüm, Unlü, Tez, & Gülsün, 1994; Güven, Ozbay, Unlü, & Satar,
1999). In many studies, fish are the subject of investigations on
heavy metal accumulations and monitoring programs in seas,
due to their importance in human nutrition (Catsiki & Strogyloudi,
1999; Keskin et al., 2007; Türkmen, Türkmen, Tepe, Töre, & Ates,
2009; Uluozlu, Tuzen, Mendil, & Soylak, 2007; Usero, Izquierdo,
Morillo, & Gracia, 2003). Data show that metal concentrations in
fish muscles vary widely, depending on the location of capture.
Several studies have shown that the accumulation of heavy metals
in tissues is primarily dependent upon the needs, sex, size and molt
* Corresponding author. Tel.: +385 1 612 3601; fax: +385 1 612 3636.
E-mail address: bilandzic@veinst.hr (N. Bilandžić).
of marine animals and also upon the water concentrations of met-
als and the exposure period, while other environmental factors
such as salinity, pH, hardness and temperature also play significant
roles in metal accumulation (Bryan & Langston, 1992; Canli & Atli,
2003; Kalay, Ay, & Canli, 1999; Zhang & Wong, 2007). Previous re-
ports have indicated that marine organisms contain much higher
concentrations of arsenic than freshwater species (Fattorini et al.,
2008). Therefore, higher As levels were found in mussel tissues
from the sea around the cities of Šibenik and Split in Croatia and
in off-shore districts in the northern and central Adriatic Sea
(Fattorini et al., 2008; Orescanin et al., 2006) than in fish species
from the Neretva River (Has-Schon, Bogut, & Strelec, 2006).
Mercury, lead, cadmium and arsenic are considered to be the
most dangerous metals and have been associated with serious ad-
verse health effects. Copper, although an essential metal, may pro-
duce toxic effects such as liver cirrhosis, dermatitis and
neurological disorders when consumed in excessive amounts in
foods such as fish and fish products (Storelli, Barone, Garofalo, &
Marcotrigiano, 2007). The most common toxic effects of metals in-
volve the brain and kidneys. Mercury is one of the most toxic heavy
metals in the environment and it is a well-known fact that the gen-
eral population is most commonly exposed to mercury through the
consumption of fish and marine mammals that may contain
methyl-mercury in their tissues (Castro-Gonzáleza & Méndez-
Armenta, 2008). Lead exposure can cause a wide spectrum of
health problems, ranging from convulsions, coma and renal failure.
Acute high dose cadmium exposure can cause severe respiratory
irritation, while long-term cadmium exposure has proven to be a

0308-8146/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.07.060
1006 N. Bilandžić et al. / Food Chemistry 124 (2011) 1005–1010
risk factor for chronic lung disease and testicular degeneration
(Benoff, Jacop, & Hurley, 2000). Arsenic exposure has been related
to the appearance of cancers in humans, including lung, liver, skin
and bladder cancer (Kapaj, Peterson, Liber, & Bhattacharya, 2006).
Several studies report heavy metal concentrations in fish from
the Croatian (Kljaković Gašparić, Zvonarić, Vrgoč, Odžak, & Barić,
2002) and Italian waters (Sepe et al., 2003) of the Adriatic Sea.
However, there are limited data on the heavy metal content in fish
from the Adriatic Sea, especially in those fish species that are an
important component of the human diet along the Croatian coast.
Therefore, the objective of this study was to determine the accu-
mulation of As, Cu, Cd, Hg and Pb in four fish species commonly
caught in the Adriatic Sea: anchovy (Engraulis encrasicholus), mack-
erel (Scomber japonicus), red mullet (Mullus surmuletus) and picarel
(Spicara smaris). Accordingly, the metal pollution concentrations of
these four species were used to determine whether they are suit-
able for human consumption.
2. Materials and methods
2.1. Sample collection
Fresh fish samples were obtained at fish markets in Croatian
coastal cities from commercial fishing efforts of local fishermen
Fig. 1. The most common locations of sampling locations from the coastal waters of Croatia.
in coastal and open waters of the eastern Adriatic Sea during
2008 and 2009 (Fig. 1). Sample collection was organized within
the National Residue Monitoring Plan of the Republic of Croatia.
Specimens collected during the sampling period were separated
into four fish species: 45 anchovy (E. encrasicholus), 38 mackerel
(S. japonicus) , 37 red mullet ( M. surmuletus) and 33 picarel
( S. smaris). Following collection, specimens were frozen in pre-
washed polyethylene bags, brought to the laboratory, and stored
frozen at 18 °C prior to analysis.
2.2. Reagents
All reagents were of analytical reagent grade, HNO3, H2O2, and
HCl (Analytical Grade, Kemika, Croatia). Double deionised water
(Milli-Q Millipore, 18.2 MX cm 1 resistivity) was used for all dilu-
tions. All plastic and glassware were cleaned by soaking in diluted
HNO3 (1/9, v/v) and rinsed with distilled water prior to use. Cali-
brations were prepared with element standard solutions of
1000 mg l 1 of each element supplied by Perkin Elmer. Stock solu-
tion was diluted in HNO3 (0.2%). As matrix modifiers in each atom-
ization for Pb and Cd, 0.005 mg Pd(NO3)2 and 0.003 mg Mg(NO3)2
(Perkin Elmer, USA) were used. The hydride technique for mercury
determinations involves the reaction of acidified aqueous samples
(3% v/v HCl) with a reducing agent 0.2% sodium borohydride in
0.05% NaOH.
 N. Bilandžić et al. / Food Chemistry 124 (2011) 1005–1010
2.3. Apparatus
Analyses of As, Cd, Cu and Pb were conducted by graphite fur-
nace-atomic absorption spectroscopy using an AAnalyst 800 atom-
ic absorption spectrometer (Perkin Elmer, USA) equipped with an
AS 800 autosampler (Perkin Elmer, USA). For graphite furnace mea-
surements, argon was used as the inert gas. Pyrolytic-coated
graphite tubes with a platform were used. The atomic absorption
signal was measured in peak area mode against a calibration curve.
Mercury was analyzed by the cold vapour technique with a flow
injection system coupled to an FIAS-100 atomic absorption spec-
trophotometer (Perkin Elmer, USA) equipped with an AS 93 plus
autosampler (Perkin Elmer). Microwave closed system Multiwave
3000 (Anton Paar, Germany) was used for digestion of samples.
The operating parameters for working elements are presented in
Table 1.
2.4. Microwave digestion
Samples (2 g) were digested with 5 ml of HNO3 (65% v/v), 1 ml
of H2O2 (30% v/v) with a microwave oven. A blank digest was car-
ried out in the same way. The digestion program began at a po-
tency of 1200 W then ramped for 10 min, after which samples
were held for 10 min at 1200 W. The second step began at a po-
tency of 0 W and held for 15 min. Digested samples were diluted
to a final volume of 50 ml with double deionised water. All metal
concentrations were determined on a wet weight basis as mg kg 1.
Detection limits were determined as the concentration corre-
sponding to three times the standard deviation of ten blanks. All
specimens were run in batches that included blanks, a standard
calibration curve, two spiked specimens, and one duplicate. In or-
der to validate the method for accuracy and precision, certified ref-
erence materials of dogfish muscle (DORM-2, National Research
Council, Canada) were analyzed for corresponding elements.
2.5. Statistical analyses
Statistical analysis was performed using the StatisticaÒ 6.1
(StatSoftÒ Inc., Tulsa, USA). Data were grouped according to spe-
cies. One-way analysis of variance was used to test for differences
in tissue metal concentrations. Data were log-transformed to im-
prove normality before analysis to meet the underlying assump-
tions of the analysis of variance; the values given are therefore
geometric means. The differences between the metal concentra-
tions in different species were analyzed using the t-test. Possibili-
ties less than 0.05 were considered statistically significant
(p < 0.05).
3. Results and discussion
The accuracy of the method was evaluated by means of trace
metal determination in the certified reference material. The
achieved recovery rates were in good agreement with certified val-
Table 1
Instrumental analytical condition of studied elements.
Element Wavelength Argon flow Sample volume
(nm) (ml min 1) (ll)
Condition for graphite furnace-atomic absorption spectrometry and flow injection
analysis system (for Hg)
Pb 283.3 250 20
Cd 228.8 250 20
Cu 324.8 250 20
As 193.7 250 20
Hg 253.7 50 500
1007
Table 2
Concentrations of metals (means ± standard errors, in mg kg 1 dry wt, n = 5) found in
certified references material DORM-2 (dogfish muscle, NRC, Canada).
Element Certified values Measured values Recovery (%)
Cd 0.043 ± 0.008 0.041 ± 0.011 95.4
Cu 2.34 ± 0.16 2.31 ± 0.24 98.7
Hg 4.64 ± 0.26 4.55 ± 0.69 98.1
Pb 0.065 ± 0.007 0.068 ± 0.009 104.6
As 18 ± 1.1 16.8 ± 2.23 93.3
ues (Table 2). The limits of detection (LODs, mg kg 1) of elements
were found to be: As 0.01, Cd 0.0004, Cu 0.0005, Hg 0.0004 and
Pb 0.005.
Concentrations of five elements (geometric means and range) in
the muscles of four fish species from Croatian coastal waters of the
Adriatic Sea, i.e. anchovy (E. encrasicholus), mackerel (S. japonicus),
red mullet ( M. surmuletus) and picarel ( S. smaris), are shown in
Table 3. Statistical analysis using the one-way ANOVA showed a
significant difference in arsenic (p < 0.0001), cadmium (p <
0.0001) and copper (p < 0.001) levels for the fish species observed.
There were no significant differences in lead and mercury levels
among the species. Differences observed by the t-test in the mean
metal values obtained for four fish species are presented in Table 3.
Arsenic concentrations ranged from minimum values of
0.01 mg kg 1 to: 54.8 mg kg 1 for anchovy, 36.4 mg kg 1 for mack-
erel, 70.9 mg kg 1 for red mullet and 54.6 mg kg 1 for picarel. The
highest and lowest mean arsenic concentration of 5.91 mg kg 1
was found in red mullet and 0.43 mg kg 1 in anchovy. The maxi-
mum arsenic level permitted for marine fish is 2 mg kg 1, accord-
ing to the guidelines of the European Community (EEC, 2001) and
the Croatian legislation (Ordinance, 2008). In this study, the mean
arsenic level found in red mullet was higher (5.91 mg kg 1) than
the prescribed limit. However, in Catalonia, Spain, lower levels of
arsenic were reported (mg kg 1) in the following ranges: 15.39–
17.77 in red mullet, 3.93–5.42 in anchovy and 1.73–7.47 in mack-
erel (Falcó, Llobet, Bocio, & Domingo, 2006). In three fish species
from the southern Atlantic coast of Spain, arsenic concentration
ranged from 0.52 to 3.96 mg kg 1 and the mean metal concentra-
tions in different fish showed significant differences (Usero et al.,
2003). Arsenic levels found were higher than fish muscle concen-
trations reported in fish from the Black Sea in Turkey, where they
ranged from 0.11 to 0.32 mg kg 1 (Tuzen, 2009). In fish from the
Gulf of Mexico, the mean arsenic concentration was 7.0 mg kg 1
(Lewis et al., 2002). Arsenic accumulation in different muscle tis-
sues of 10 fish species in Manchar Lake, Pakistan that are com-
monly consumed by the local population showed significant
differences in arsenic concentrations, ranging between 2.0 and
14.8 mg kg 1 (Shah et al., 2009). In previous studies, As levels mea-
sured in the soft tissue of Mediterranean blue mussel (Mytilus
galloprovincialis) sampled in the unpolluted sea around the city of
Šibenik (Most and Luka) and polluted waters around the city of
Split on the Croatian coast (Marina, Jugovinil, Vranjic and Stobrec)
showed variation in the range from 4 to 30 mg kg 1, with a mean
value of 15.2 mg kg 1 (Orescanin et al., 2006). No significant differ-
ences between study areas were observed. Results obtained in the
study of mussel tissues of Mytilus galloprovincialis, sampled over a
five year period from 41 platforms in the central Adriatic Sea (Por-
Modifier
(ll) tonovo, Ancona, Italy) shown seasonal, inter-annual and geograph-
ical variability of As concentrations, ranging between 10 and
20 mg kg 1 (Fattorini et al., 2008). Investigations off the US coast
5 suggested that environmental factors such as the seasonal cycle
5 of absorption/solubilization of the element in specific observed
5 areas, local physico-chemical parameters such as temperature,
5
salinity and the nature of sediments might affect the large bioaccu-
–
mulation of As (Valette-Silver et al., 1999). In fact, the different
1008 N. Bilandžić et al. / Food Chemistry 124 (2011) 1005–1010
Table 3
Metal concentrations (geometric mean mg kg 1 wet wt and range) in the muscle tissues of the anchovy (Engraulis encrasicholus), mackerel (Scomber japonicus), red mullet (Mullus
surmuletus) and picarel (Spicara smaris) from the Adriatic Sea, Croatia.
Species N Arsenic
Anchovy (Engraulis encrasicholus) 45 Geometric mean 0.43c
Range (min–max) 0.01–54.8
Mackerel (Scomber japonicus) 38 Geometric mean 0.53b
Range (min–max) 0.01–36.4
Red mullet (Mullus surmuletus) 37 Geometric mean 5.91bc
Range (min–max) 0.01–70.9
Picarel (Spicara smaris) 33 Geometric mean 0.72b
Range (min–max) 0.01–54.6
Vertically, letters show statistically significant differences obtained by t-test: a (p < 0.05); b (p < 0.01); c (p < 0.0001).
levels of As measured in mussels sampled from the off-shore dis-
tricts of the northern and central Adriatic Sea are due to the signif-
icant influence of seawater salinity in modulating accumulation of
the metal (Fattorini et al., 2008). Previous findings have deter-
mined the natural origin of As, which was mostly present as arsen-
obetaine, a non-toxic arsenic compound normally accumulated by
marine organisms through diet and not released from anthropo-
genic activities (Fattorini et al., 2004).
Copper levels in analysed fish were in the ranges: 0.001–
57.3 mg kg 1 in red mullet, 0.001–6.29 mg kg 1 in anchovy, 0.001–
15.9 mg kg 1 in mackerel and 0.081–32.9 mg kg 1 in picarel. The
lowest and highest mean copper concentrations in fish species
were 0.13 mg kg 1 in mackerel and 1.47 mg kg 1 in picarel. The
Croatian legislation has not determined the maximum permitted
copper level. However, the maximum permitted copper level
established by the FAO (1983) and WHO (1996) is 30 mg kg 1
and 20 mg kg 1 by MAFF (1995). The FAO/WHO has set a provi-
sional tolerable daily intake (PTDI, of 3 mg of copper based on
the body weight for an average adult (60 kg body weight) (Joint
FAO/WHO Expert Committee on Food Additives, 1999). Mean cop-
per levels in analyzed fish samples were found to be lower than
these limits. Copper levels presented in this study were similar
to fish muscle concentrations reported in the northeast Atlantic,
ranging between 0.11 and 0.97 mg kg 1 (Celik & Oehlenschlager,
2004), and in Spain, ranging between 0.4 and 1.5 mg kg 1 (Usero
et al., 2003), in Tuzla Lagoon, Turkey, ranging between 0.26 and
0.82 mg kg 1 (Dural, Goksu, & Ozak, 2007), and in the Black and
Aegean Seas, ranging between 0.73 and 1.83 mg kg 1 (Uluozlu
et al., 2007). However, copper levels found were lower than levels
reported in the Black Sea, ranging between 1.28 and 2.93 mg kg 1,
0.65 and 2.78 mg kg 1 and 1.4 and 2.4 mg kg 1 (Tuzen, 2003, 2009;
Mendil, Demirci, Tuzen, & Soylak, 2010); in Turkey in the northeast
Mediterranean Sea, ranging from 0.65 to 4.36 mg kg 1 (Yilmaz,
Ozdemir, Demirak, & Tuna, 2007) and from 0.639 to 2.976 mg kg 1
(Turkmen, Turkmen, Tepe, & Akyurt, 2005); in Marmara Sea, rang-
ing from 0.23 to 9.49 mg kg 1 (Keskin et al., 2007) and in the lakes
in Tokrat ranging from 1.0 to 4.1 mg kg 1 (Mendil et al., 2005).
Also, similar levels of copper ranging from 1.1 to 2.5 mg kg 1 were
reported in canned fish in Turkey (Tuzen & Soylak, 2007). Copper
levels in mackerel muscle measured in the Aegean and Mediterra-
nean Seas ranged in different locations from 0.51 to 6.22 mg kg 1
(Türkmen et al., 2009). In six fish species caught in the northeast
Mediterranean Sea, copper levels in muscle tissue ranged from
2.19 to 4.41 mg kg 1 (Canli & Atli, 2003).
Cadmium levels in muscles of fish species ranged: 0.002–
0.85 mg kg 1 in red mullet, 0.001–0.02 mg kg 1 in anchovy, 0.001–
0.1 mg kg 1 in mackerel and 0.001–0.97 mg kg 1 in picarel. The
mean lowest cadmium content 0.002 mg kg 1 was found in ancho-
vy and red mullet while the highest cadmium level was 0.006
mg kg 1 in red mullet. Cadmium mean levels in the analysed fish
samples (mg kg 1) were below the maximum permissible value
Cadmium Copper Lead Mercury
0.002bc 0.23ab 0.01 0.04
0.001–0.02 0.001–6.29 0.001–0.34 0.001–0.52
0.006bc 0.13b 0.01 0.08
0.001–0.1 0.001–15.9 0.002–0.24 0.001–0.78
0.002b 0.79a 0.02 0.06
0.001–0.85 0.001–57.6 0.001–0.27 0.001–2.07
0.003b 1.47b 0.02 0.08
0.001–0.097 0.081–32.9 0.001–0.46 0.001–2.06
indicated by the European Community (EEC, 2001) and the
Croatian legislation (Ordinance, 2008): 0.3 for Engraulis sp., 0.1
for Scomber sp. and 0.05 for red mullet (M. surmuletus) and picarel
( S. smaris). The maximum cadmium level permitted by the FAO
(1983) is 0.5 mg kg 1 and 0.2 mg kg 1 by MAFF (1995). The cad-
mium levels found in red mullet muscle in this study were higher
than levels obtained in previous investigation in red mullet from
the eastern Adriatic Sea, ranging from 0.007 to 0.029 mg kg 1
(Kljaković Gašparić et al., 2002). Also, the highest values (mg kg 1)
were observed, mainly in the central area of the Adriatic Sea: an-
chovy 0.020, red mullet 0.003 and mackerel 0.008 (Sepe et al.,
2003). In fish species in Catalonia, Spain, cadmium levels were
0.001–0.01 mg kg 1 in red mullet, 0.001–0.02 mg kg 1 in anchovy,
0.005–0.01 mg kg 1 in hake and 0.003–0.01 mg kg 1 in mackerel
(Falcó et al., 2006). Also, in six fish species caught from the north-
east Mediterranean Sea concentrations of cadmium in muscle tis-
sue ranged from 0.37 to 0.79 mg kg 1 (Canli & Atli, 2003).
However, cadmium levels found in anchovy, picarel and mackerel
muscles in this study are lower than in previous studies reported in
different sea locations from Turkey in different fish species, rang-
ing (mg kg 1) from 0.01 to 4.16 (Türkmen et al., 2005) and 0.010
to 0.084 in Iskenderun Bay (Yilmaz et al., 2007), 0.09 to 0.48, 0.1
to 0.35 and 0.18 to 0.35 in the Black Sea (Mendil et al., 2010; Tuzen,
2003, 2009), <0.02 to 0.24 off the Black Sea coasts (Topcuoglu,
Kırbasoglu, & Güngor, 2002), 0.45 to 0.80 in the Black and Aegean
Seas (Uluozlu et al., 2007), 0.03 to 0.12 in Tuzla Lagoon, Mediterra-
nean Sea region (Dural et al., 2007), 0.03 to 0.39 in the Aegean and
Mediterranean Seas (Türkmen et al., 2009) and 0.2 to 1.2 in the
lakes in Tokrat (Mendil et al., 2005). Similar results in the range
0.06–0.25 mg kg 1 were reported in canned fish marketed in
Turkey (Tuzen & Soylak, 2007).
In the present study, the minimum and maximum lead concen-
tration in muscle tissues (mg kg 1) were 0.001 and 0.27 in red mul-
let, 0.001 and 0.34 in anchovy, 0.002 and 0.24 in mackerel and
0.001 and 0.46 in picarel. Mean lead levels in the analysed fish
samples (0.01–0.02 mg kg 1) were 14–30 times lower than the
maximum permissible value of 0.3 mg kg 1 determined by the
European Community (EEC, 2001) and the Croatian legislation
(Ordinance, 2008). The maximum lead level permitted determined
by the WHO (1996) is 2.0 mg kg 1, 0.5 mg kg 1 by the FAO (1983)
and 2.0 mg kg 1 by the MAFF (1995). The FAO/WHO has set 214 lg
as the provisional tolerable daily intakes (PTDI) for lead for an
average adult of 60 kg body weight (Joint FAO/WHO Expert Com-
mittee on Food Additives, 1999). Lead levels found were lower than
those reported in fish from the eastern Adriatic in the range 0.049–
0.141 mg kg 1 in hake and 0.057–0.158 mg kg 1 in red mullet
(Kljaković Gašparić et al., 2002). In six fish species collected along
the coast of the Adriatic Sea, the highest lead values (mg kg 1)
were observed in the central area of the Adriatic Sea, for anchovy
0.046 mg kg 1, red mullet 0.036 mg kg 1 and mackerel 0.011 mg
kg 1 (Sepe et al., 2003). Lead levels found in fish from Catalonia,
 N. Bilandžić et al. / Food Chemistry 124 (2011) 1005–1010
Spain, ranged (mg kg 1) between 0.002 and 0.07 in red mullet, 0.01
and 0.02 in anchovy, 0.01 and 0.02 in mackerel (Falcó et al., 2006).
However, lead concentrations measured in Turkey, Northern Med-
iterranean Sea, ranged from 0.09 to 6.95 mg kg 1 (Türkmen et al.,
2005), 0.22 to 0.85, 0.28 to 0.87 and 0.28 to 0.64 mg kg 1 in fish
samples from the Black Sea (Mendil et al., 2010; Tuzen, 2003,
2009), 0.068 to 0.874 mg kg 1 (Yilmaz et al., 2007) and 0.33 to
0.93 mg kg 1 in the Black and Aegean Seas (Uluozlu et al., 2007),
0.49 to 2.44 mg kg 1 from the Tuzla Lagoon (Dural et al., 2007),
0.14 to 1.28 mg kg 1 in the Aegean and Mediterranean Seas
(Türkmen et al., 2009), and 0.7 to 2.4 mg kg 1 in the lakes in Tokrat
(Mendil et al., 2005). On the other hand, lead levels reported in
canned fish in Turkey ranged from 0.09 to 0.4 mg kg 1 (Tuzen &
Soylak, 2007). However, in six fish from the northeast Mediterra-
nean Sea, lead levels were much higher and ranged from 2.98 to
6.12 mg kg 1 (Canli & Atli, 2003).
Mercury levels (mg kg 1) in fish muscles in this study ranged
from a minimum of 0.001 to a maximum of 2.07 in red mullet,
0.52 in anchovy, 0.78 in mackerel and 2.06 in picarel. However,
mean metal levels in the analysed fish samples (0.04–0.08
mg kg 1) were below the maximum permissible value indicated
by the European Community (EEC, 2001) and the Croatian legisla-
tion (Ordinance, 2008), i.e. 0.5 mg kg 1 for Engraulis sp., Scomber sp.
and Spicara sp. and 1.0 mg kg 1 for red mullet (Mullus surmuletus).
MAFF (1995) has established a maximum permitted mercury level
of 0.5 mg kg 1 in fish. In a recent study of fish species from the
Black Sea in Turkey, similar mercury levels were reported and ran-
ged from 0.025 to 0.078 mg kg 1 (Tuzen, 2009). In Spain, muscle
mercury concentrations ranged from 0.14 to 0.36 mg kg 1 in red
mullet, 0.08 to 0.09 mg kg 1 in anchovy, 0.12 to 0.29 mg kg 1 in
hake, and 0.06 to 0.15 mg kg 1 in mackerel (Falcó et al., 2006).
In the southern Atlantic coast of Spain, mercury concentration in
three fish species ranged from 0.01 to 0.023 mg kg 1 (Usero
et al., 2003). Higher levels of mercury (0.39 mg kg 1) were found
in hake of the Adriatic Sea (Storelli, Storelli, Giacominelli-Stuffler,
& Marcotrigiano, 2005) and black scabbard fish (0.90 mg kg 1)
caught in Portugal (Afonso, Lourenc, Abreu Dias, Nunes, & Castro,
2007).
In conclusion, statistically significant differences were observed
in mean metal values obtained for fish species from the Adriatic
Sea. The results presented on metal contents in the examined spe-
cies gives an indication of the environmental conditions along the
eastern coast of the Adriatic Sea. Accordingly, the concentrations of
Cd, Cu, Hg and Pb obtained were far below the established values
by the European Community regulations. However, arsenic levels
found in red mullet were higher than the recommended legal lim-
its for human consumption and may present a human health issue.
Acknowledgements
The authors are thankful to all those who contributed to the
study. Special thanks go to Marijana Fluka and Mirjana Hren for
sample preparation. This study was supported with a grant from
the Ministry of Agriculture, Fisheries and Rural Development Vet-
erinary Directorate, Croatia.
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