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 of marine animals and also upon the water concentrations of met-
als and the exposure period, while other environmental factors
Heavy metal toxins have been present in every segment of hu- such as salinity, pH, hardness and temperature also play significant
man life for an immeasurable amount of time due to our increased roles in metal accumulation (Bryan & Langston, 1992; Canli & Atli,
use of such compounds. Heavy metal toxins can induce a variety of 2003; Kalay, Ay, & Canli, 1999; Zhang & Wong, 2007). Previous re-
adverse health effects in an exposed individual, such as behav- ports have indicated that marine organisms contain much higher
ioural, physiological, and cognitive effects, depending on the type concentrations of arsenic than freshwater species (Fattorini et al.,
of toxin. Heavy metals are discharged into the marine environment 2008). Therefore, higher As levels were found in mussel tissues
through urban discharge, agriculture, mining, combustion and from the sea around the cities of Šibenik and Split in Croatia and
industrial discharge, and can remain in solution or in suspension in off-shore districts in the northern and central Adriatic Sea
and precipitate to the bottom, or be taken up by organisms, thus (Fattorini et al., 2008; Orescanin et al., 2006) than in fish species
creating a potential source of heavy metal pollution in the aquatic from the Neretva River (Has-Schon, Bogut, & Strelec, 2006).
environment. Accordingly, metals can accumulate to toxic concen- Mercury, lead, cadmium and arsenic are considered to be the
trations and cause ecological damage, and subsequently be trans- most dangerous metals and have been associated with serious ad-
ferred to humans through the food chain (Giordano et al., 1991; verse health effects. Copper, although an essential metal, may pro-
Gümgüm, Unlü, Tez, & Gülsün, 1994; Güven, Ozbay, Unlü, & Satar, duce toxic effects such as liver cirrhosis, dermatitis and
1999). In many studies, fish are the subject of investigations on neurological disorders when consumed in excessive amounts in
heavy metal accumulations and monitoring programs in seas, foods such as fish and fish products (Storelli, Barone, Garofalo, &
due to their importance in human nutrition (Catsiki & Strogyloudi, Marcotrigiano, 2007). The most common toxic effects of metals in-
1999; Keskin et al., 2007; Türkmen, Türkmen, Tepe, Töre, & Ates, volve the brain and kidneys. Mercury is one of the most toxic heavy
2009; Uluozlu, Tuzen, Mendil, & Soylak, 2007; Usero, Izquierdo, metals in the environment and it is a well-known fact that the gen-
Morillo, & Gracia, 2003). Data show that metal concentrations in eral population is most commonly exposed to mercury through the
fish muscles vary widely, depending on the location of capture. consumption of fish and marine mammals that may contain
Several studies have shown that the accumulation of heavy metals methyl-mercury in their tissues (Castro-Gonzáleza & Méndez-
in tissues is primarily dependent upon the needs, sex, size and molt Armenta, 2008). Lead exposure can cause a wide spectrum of
health problems, ranging from convulsions, coma and renal failure.
* Corresponding author. Tel.: +385 1 612 3601; fax: +385 1 612 3636. Acute high dose cadmium exposure can cause severe respiratory
E-mail address: [email protected] (N. Bilandžić). 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 in coastal and open waters of the eastern Adriatic Sea during
(Benoff, Jacop, & Hurley, 2000). Arsenic exposure has been related 2008 and 2009 (Fig. 1). Sample collection was organized within
to the appearance of cancers in humans, including lung, liver, skin the National Residue Monitoring Plan of the Republic of Croatia.
and bladder cancer (Kapaj, Peterson, Liber, & Bhattacharya, 2006). Specimens collected during the sampling period were separated
Several studies report heavy metal concentrations in fish from into four fish species: 45 anchovy (E. encrasicholus), 38 mackerel
the Croatian (Kljaković Gašparić, Zvonarić, Vrgoč, Odžak, & Barić, (S. japonicus) , 37 red mullet ( M. surmuletus) and 33 picarel
2002) and Italian waters (Sepe et al., 2003) of the Adriatic Sea. ( S. smaris). Following collection, specimens were frozen in pre-
However, there are limited data on the heavy metal content in fish washed polyethylene bags, brought to the laboratory, and stored
from the Adriatic Sea, especially in those fish species that are an frozen at 18 °C prior to analysis.
important component of the human diet along the Croatian coast.
Therefore, the objective of this study was to determine the accu- 2.2. Reagents
mulation of As, Cu, Cd, Hg and Pb in four fish species commonly
caught in the Adriatic Sea: anchovy (Engraulis encrasicholus), mack- All reagents were of analytical reagent grade, HNO3, H2O2, and
erel (Scomber japonicus), red mullet (Mullus surmuletus) and picarel HCl (Analytical Grade, Kemika, Croatia). Double deionised water
(Spicara smaris). Accordingly, the metal pollution concentrations of (Milli-Q Millipore, 18.2 MX cm 1 resistivity) was used for all dilu-
these four species were used to determine whether they are suit- tions. All plastic and glassware were cleaned by soaking in diluted
able for human consumption. 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-
2. Materials and methods 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
2.1. Sample collection (Perkin Elmer, USA) were used. The hydride technique for mercury
determinations involves the reaction of acidified aqueous samples
Fresh fish samples were obtained at fish markets in Croatian (3% v/v HCl) with a reducing agent 0.2% sodium borohydride in
coastal cities from commercial fishing efforts of local fishermen 0.05% NaOH.

Fig. 1. The most common locations of sampling locations from the coastal waters of Croatia.
 N. Bilandžić et al. / Food Chemistry 124 (2011) 1005–1010 1007

2.3. Apparatus 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).
Analyses of As, Cd, Cu and Pb were conducted by graphite fur-
nace-atomic absorption spectroscopy using an AAnalyst 800 atom- Element Certified values Measured values Recovery (%)
ic absorption spectrometer (Perkin Elmer, USA) equipped with an Cd 0.043 ± 0.008 0.041 ± 0.011 95.4
AS 800 autosampler (Perkin Elmer, USA). For graphite furnace mea- Cu 2.34 ± 0.16 2.31 ± 0.24 98.7
surements, argon was used as the inert gas. Pyrolytic-coated Hg 4.64 ± 0.26 4.55 ± 0.69 98.1
Pb 0.065 ± 0.007 0.068 ± 0.009 104.6
graphite tubes with a platform were used. The atomic absorption As 18 ± 1.1 16.8 ± 2.23 93.3
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 ues (Table 2). The limits of detection (LODs, mg kg 1) of elements
autosampler (Perkin Elmer). Microwave closed system Multiwave were found to be: As 0.01, Cd 0.0004, Cu 0.0005, Hg 0.0004 and
3000 (Anton Paar, Germany) was used for digestion of samples. Pb 0.005.
The operating parameters for working elements are presented in Concentrations of five elements (geometric means and range) in
Table 1. the muscles of four fish species from Croatian coastal waters of the
Adriatic Sea, i.e. anchovy (E. encrasicholus), mackerel (S. japonicus),
2.4. Microwave digestion red mullet ( M. surmuletus) and picarel ( S. smaris), are shown in
Table 3. Statistical analysis using the one-way ANOVA showed a
Samples (2 g) were digested with 5 ml of HNO3 (65% v/v), 1 ml significant difference in arsenic (p < 0.0001), cadmium (p <
of H2O2 (30% v/v) with a microwave oven. A blank digest was car- 0.0001) and copper (p < 0.001) levels for the fish species observed.
ried out in the same way. The digestion program began at a po- There were no significant differences in lead and mercury levels
tency of 1200 W then ramped for 10 min, after which samples among the species. Differences observed by the t-test in the mean
were held for 10 min at 1200 W. The second step began at a po- metal values obtained for four fish species are presented in Table 3.
tency of 0 W and held for 15 min. Digested samples were diluted Arsenic concentrations ranged from minimum values of
to a final volume of 50 ml with double deionised water. All metal 0.01 mg kg 1 to: 54.8 mg kg 1 for anchovy, 36.4 mg kg 1 for mack-
concentrations were determined on a wet weight basis as mg kg 1. erel, 70.9 mg kg 1 for red mullet and 54.6 mg kg 1 for picarel. The
Detection limits were determined as the concentration corre- highest and lowest mean arsenic concentration of 5.91 mg kg 1
sponding to three times the standard deviation of ten blanks. All was found in red mullet and 0.43 mg kg 1 in anchovy. The maxi-
specimens were run in batches that included blanks, a standard mum arsenic level permitted for marine fish is 2 mg kg 1, accord-
calibration curve, two spiked specimens, and one duplicate. In or- ing to the guidelines of the European Community (EEC, 2001) and
der to validate the method for accuracy and precision, certified ref- the Croatian legislation (Ordinance, 2008). In this study, the mean
erence materials of dogfish muscle (DORM-2, National Research arsenic level found in red mullet was higher (5.91 mg kg 1) than
Council, Canada) were analyzed for corresponding elements. the prescribed limit. However, in Catalonia, Spain, lower levels of
arsenic were reported (mg kg 1) in the following ranges: 15.39–
2.5. Statistical analyses 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
Statistical analysis was performed using the StatisticaÒ 6.1 from the southern Atlantic coast of Spain, arsenic concentration
(StatSoftÒ Inc., Tulsa, USA). Data were grouped according to spe- ranged from 0.52 to 3.96 mg kg 1 and the mean metal concentra-
cies. One-way analysis of variance was used to test for differences tions in different fish showed significant differences (Usero et al.,
in tissue metal concentrations. Data were log-transformed to im- 2003). Arsenic levels found were higher than fish muscle concen-
prove normality before analysis to meet the underlying assump- trations reported in fish from the Black Sea in Turkey, where they
tions of the analysis of variance; the values given are therefore ranged from 0.11 to 0.32 mg kg 1 (Tuzen, 2009). In fish from the
geometric means. The differences between the metal concentra- Gulf of Mexico, the mean arsenic concentration was 7.0 mg kg 1
tions in different species were analyzed using the t-test. Possibili- (Lewis et al., 2002). Arsenic accumulation in different muscle tis-
ties less than 0.05 were considered statistically significant sues of 10 fish species in Manchar Lake, Pakistan that are com-
(p < 0.05). 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-
3. Results and discussion sured in the soft tissue of Mediterranean blue mussel (Mytilus
galloprovincialis) sampled in the unpolluted sea around the city of
The accuracy of the method was evaluated by means of trace Šibenik (Most and Luka) and polluted waters around the city of
metal determination in the certified reference material. The Split on the Croatian coast (Marina, Jugovinil, Vranjic and Stobrec)
achieved recovery rates were in good agreement with certified val- 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-
Table 1 ences between study areas were observed. Results obtained in the
Instrumental analytical condition of studied elements. study of mussel tissues of Mytilus galloprovincialis, sampled over a
five year period from 41 platforms in the central Adriatic Sea (Por-
Element Wavelength Argon flow Sample volume Modifier
(nm) (ml min 1) (ll) (ll) tonovo, Ancona, Italy) shown seasonal, inter-annual and geograph-
ical variability of As concentrations, ranging between 10 and
Condition for graphite furnace-atomic absorption spectrometry and flow injection
analysis system (for Hg)
20 mg kg 1 (Fattorini et al., 2008). Investigations off the US coast
Pb 283.3 250 20 5 suggested that environmental factors such as the seasonal cycle
Cd 228.8 250 20 5 of absorption/solubilization of the element in specific observed
Cu 324.8 250 20 5 areas, local physico-chemical parameters such as temperature,
As 193.7 250 20 5
salinity and the nature of sediments might affect the large bioaccu-
Hg 253.7 50 500 –
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 Cadmium Copper Lead Mercury

Anchovy (Engraulis encrasicholus) 45 Geometric mean 0.43c 0.002bc 0.23ab 0.01 0.04
Range (min–max) 0.01–54.8 0.001–0.02 0.001–6.29 0.001–0.34 0.001–0.52
Mackerel (Scomber japonicus) 38 Geometric mean 0.53b 0.006bc 0.13b 0.01 0.08
Range (min–max) 0.01–36.4 0.001–0.1 0.001–15.9 0.002–0.24 0.001–0.78
Red mullet (Mullus surmuletus) 37 Geometric mean 5.91bc 0.002b 0.79a 0.02 0.06
Range (min–max) 0.01–70.9 0.001–0.85 0.001–57.6 0.001–0.27 0.001–2.07
Picarel (Spicara smaris) 33 Geometric mean 0.72b 0.003b 1.47b 0.02 0.08
Range (min–max) 0.01–54.6 0.001–0.097 0.081–32.9 0.001–0.46 0.001–2.06

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- indicated by the European Community (EEC, 2001) and the
tricts of the northern and central Adriatic Sea are due to the signif- Croatian legislation (Ordinance, 2008): 0.3 for Engraulis sp., 0.1
icant influence of seawater salinity in modulating accumulation of for Scomber sp. and 0.05 for red mullet (M. surmuletus) and picarel
the metal (Fattorini et al., 2008). Previous findings have deter- ( S. smaris). The maximum cadmium level permitted by the FAO
mined the natural origin of As, which was mostly present as arsen- (1983) is 0.5 mg kg 1 and 0.2 mg kg 1 by MAFF (1995). The cad-
obetaine, a non-toxic arsenic compound normally accumulated by mium levels found in red mullet muscle in this study were higher
marine organisms through diet and not released from anthropo- than levels obtained in previous investigation in red mullet from
genic activities (Fattorini et al., 2004). the eastern Adriatic Sea, ranging from 0.007 to 0.029 mg kg 1
Copper levels in analysed fish were in the ranges: 0.001– (Kljaković Gašparić et al., 2002). Also, the highest values (mg kg 1)
57.3 mg kg 1 in red mullet, 0.001–6.29 mg kg 1 in anchovy, 0.001– were observed, mainly in the central area of the Adriatic Sea: an-
15.9 mg kg 1 in mackerel and 0.081–32.9 mg kg 1 in picarel. The chovy 0.020, red mullet 0.003 and mackerel 0.008 (Sepe et al.,
lowest and highest mean copper concentrations in fish species 2003). In fish species in Catalonia, Spain, cadmium levels were
were 0.13 mg kg 1 in mackerel and 1.47 mg kg 1 in picarel. The 0.001–0.01 mg kg 1 in red mullet, 0.001–0.02 mg kg 1 in anchovy,
Croatian legislation has not determined the maximum permitted 0.005–0.01 mg kg 1 in hake and 0.003–0.01 mg kg 1 in mackerel
copper level. However, the maximum permitted copper level (Falcó et al., 2006). Also, in six fish species caught from the north-
established by the FAO (1983) and WHO (1996) is 30 mg kg 1 east Mediterranean Sea concentrations of cadmium in muscle tis-
and 20 mg kg 1 by MAFF (1995). The FAO/WHO has set a provi- sue ranged from 0.37 to 0.79 mg kg 1 (Canli & Atli, 2003).
sional tolerable daily intake (PTDI, of 3 mg of copper based on However, cadmium levels found in anchovy, picarel and mackerel
the body weight for an average adult (60 kg body weight) (Joint muscles in this study are lower than in previous studies reported in
FAO/WHO Expert Committee on Food Additives, 1999). Mean cop- different sea locations from Turkey in different fish species, rang-
per levels in analyzed fish samples were found to be lower than ing (mg kg 1) from 0.01 to 4.16 (Türkmen et al., 2005) and 0.010
these limits. Copper levels presented in this study were similar to 0.084 in Iskenderun Bay (Yilmaz et al., 2007), 0.09 to 0.48, 0.1
to fish muscle concentrations reported in the northeast Atlantic, to 0.35 and 0.18 to 0.35 in the Black Sea (Mendil et al., 2010; Tuzen,
ranging between 0.11 and 0.97 mg kg 1 (Celik & Oehlenschlager, 2003, 2009), <0.02 to 0.24 off the Black Sea coasts (Topcuoglu,
2004), and in Spain, ranging between 0.4 and 1.5 mg kg 1 (Usero Kırbasoglu, & Güngor, 2002), 0.45 to 0.80 in the Black and Aegean
et al., 2003), in Tuzla Lagoon, Turkey, ranging between 0.26 and Seas (Uluozlu et al., 2007), 0.03 to 0.12 in Tuzla Lagoon, Mediterra-
0.82 mg kg 1 (Dural, Goksu, & Ozak, 2007), and in the Black and nean Sea region (Dural et al., 2007), 0.03 to 0.39 in the Aegean and
Aegean Seas, ranging between 0.73 and 1.83 mg kg 1 (Uluozlu Mediterranean Seas (Türkmen et al., 2009) and 0.2 to 1.2 in the
et al., 2007). However, copper levels found were lower than levels lakes in Tokrat (Mendil et al., 2005). Similar results in the range
reported in the Black Sea, ranging between 1.28 and 2.93 mg kg 1, 0.06–0.25 mg kg 1 were reported in canned fish marketed in
0.65 and 2.78 mg kg 1 and 1.4 and 2.4 mg kg 1 (Tuzen, 2003, 2009; Turkey (Tuzen & Soylak, 2007).
Mendil, Demirci, Tuzen, & Soylak, 2010); in Turkey in the northeast In the present study, the minimum and maximum lead concen-
Mediterranean Sea, ranging from 0.65 to 4.36 mg kg 1 (Yilmaz, tration in muscle tissues (mg kg 1) were 0.001 and 0.27 in red mul-
Ozdemir, Demirak, & Tuna, 2007) and from 0.639 to 2.976 mg kg 1 let, 0.001 and 0.34 in anchovy, 0.002 and 0.24 in mackerel and
(Turkmen, Turkmen, Tepe, & Akyurt, 2005); in Marmara Sea, rang- 0.001 and 0.46 in picarel. Mean lead levels in the analysed fish
ing from 0.23 to 9.49 mg kg 1 (Keskin et al., 2007) and in the lakes samples (0.01–0.02 mg kg 1) were 14–30 times lower than the
in Tokrat ranging from 1.0 to 4.1 mg kg 1 (Mendil et al., 2005). maximum permissible value of 0.3 mg kg 1 determined by the
Also, similar levels of copper ranging from 1.1 to 2.5 mg kg 1 were European Community (EEC, 2001) and the Croatian legislation
reported in canned fish in Turkey (Tuzen & Soylak, 2007). Copper (Ordinance, 2008). The maximum lead level permitted determined
levels in mackerel muscle measured in the Aegean and Mediterra- by the WHO (1996) is 2.0 mg kg 1, 0.5 mg kg 1 by the FAO (1983)
nean Seas ranged in different locations from 0.51 to 6.22 mg kg 1 and 2.0 mg kg 1 by the MAFF (1995). The FAO/WHO has set 214 lg
(Türkmen et al., 2009). In six fish species caught in the northeast as the provisional tolerable daily intakes (PTDI) for lead for an
Mediterranean Sea, copper levels in muscle tissue ranged from average adult of 60 kg body weight (Joint FAO/WHO Expert Com-
2.19 to 4.41 mg kg 1 (Canli & Atli, 2003). mittee on Food Additives, 1999). Lead levels found were lower than
Cadmium levels in muscles of fish species ranged: 0.002– those reported in fish from the eastern Adriatic in the range 0.049–
0.85 mg kg 1 in red mullet, 0.001–0.02 mg kg 1 in anchovy, 0.001– 0.141 mg kg 1 in hake and 0.057–0.158 mg kg 1 in red mullet
0.1 mg kg 1 in mackerel and 0.001–0.97 mg kg 1 in picarel. The (Kljaković Gašparić et al., 2002). In six fish species collected along
mean lowest cadmium content 0.002 mg kg 1 was found in ancho- the coast of the Adriatic Sea, the highest lead values (mg kg 1)
vy and red mullet while the highest cadmium level was 0.006 were observed in the central area of the Adriatic Sea, for anchovy
mg kg 1 in red mullet. Cadmium mean levels in the analysed fish 0.046 mg kg 1, red mullet 0.036 mg kg 1 and mackerel 0.011 mg
samples (mg kg 1) were below the maximum permissible value kg 1 (Sepe et al., 2003). Lead levels found in fish from Catalonia,
 N. Bilandžić et al. / Food Chemistry 124 (2011) 1005–1010 1009

Spain, ranged (mg kg 1) between 0.002 and 0.07 in red mullet, 0.01 Canlı, M., & Atlı, G. (2003). The relationships between heavy metal (Cd, Cr, Cu, Fe, Pb,
Zn) levels and the size of six Mediterranean fish species. Environmental
and 0.02 in anchovy, 0.01 and 0.02 in mackerel (Falcó et al., 2006).
Pollution, 121(1), 129–136.
However, lead concentrations measured in Turkey, Northern Med- Castro-Gonzáleza, M. I., & Méndez-Armenta, M. (2008). Heavy metals: Implications
iterranean Sea, ranged from 0.09 to 6.95 mg kg 1 (Türkmen et al., associated to fish consumption. Environmental Toxicology and Pharmacology, 26,
2005), 0.22 to 0.85, 0.28 to 0.87 and 0.28 to 0.64 mg kg 1 in fish 263–271.
Catsiki, V. A., & Strogyloudi, E. (1999). Survey of metal levels in common fish species
samples from the Black Sea (Mendil et al., 2010; Tuzen, 2003, from Greek waters. The Science of the Total Environment, 237(238), 387–400.
2009), 0.068 to 0.874 mg kg 1 (Yilmaz et al., 2007) and 0.33 to Celik, U., & Oehlenschlager, J. (2004). Determination of zinc and copper in fish
0.93 mg kg 1 in the Black and Aegean Seas (Uluozlu et al., 2007), samples collected from Northeast Atlantic by DPSAV. Food Chemistry, 87,
343–347.
0.49 to 2.44 mg kg 1 from the Tuzla Lagoon (Dural et al., 2007), Dural, M., Goksu, M. Z. L., & Ozak, A. A. (2007). Investigation of heavy metal levels in
0.14 to 1.28 mg kg 1 in the Aegean and Mediterranean Seas economically important fish species captured from the Tuzla lagoon. Food
(Türkmen et al., 2009), and 0.7 to 2.4 mg kg 1 in the lakes in Tokrat Chemistry, 27, 521–526.
Mendil, D., Uluözlü, Ö. D., Hasdemir, E., Tüzen, M., Sari, H., & Suiçmez, M. (2005).
(Mendil et al., 2005). On the other hand, lead levels reported in Determination of trace metal levels in seven fish species in lakes in Tokat,
canned fish in Turkey ranged from 0.09 to 0.4 mg kg 1 (Tuzen & Turkey. Food Chemistry, 90, 175–179.
Soylak, 2007). However, in six fish from the northeast Mediterra- Mendil, D., Demirci, Z., Tuzen, M., & Soylak, M. (2010). Seasonal investigation of
trace element contents in commercially valuable fish species from the Black sea,
nean Sea, lead levels were much higher and ranged from 2.98 to Turkey. Food and Chemical Toxicology, 48, 865–870.
6.12 mg kg 1 (Canli & Atli, 2003). EEC (2001). Commission Regulation (EC) No 466/2001of 8 March 2001 setting
Mercury levels (mg kg 1) in fish muscles in this study ranged maximum levels for certain contaminants in foodstuffs. Official Journal of the
European Communities, L77.
from a minimum of 0.001 to a maximum of 2.07 in red mullet,
Falcó, G., Llobet, J. M., Bocio, A., & Domingo, J. L. (2006). Daily intake of arsenic,
0.52 in anchovy, 0.78 in mackerel and 2.06 in picarel. However, cadmium, mercury, and lead by consumption of edible marine species. Journal
mean metal levels in the analysed fish samples (0.04–0.08 of Agriculture and Food Chemistry, 54, 6106–6112.
mg kg 1) were below the maximum permissible value indicated FAO (1983). Compilation of legal limits for hazardous substances in fish and fishery
products. FAO Fishery Circular, 464, 5–10.
by the European Community (EEC, 2001) and the Croatian legisla- Fattorini, D., Alonso-Hernandez, C. M., Diaz-Asencio, M., Munoz-Caravaca, A.,
tion (Ordinance, 2008), i.e. 0.5 mg kg 1 for Engraulis sp., Scomber sp. Pannacciulli, F. G., Tangherlini, M., et al. (2004). Chemical speciation of
and Spicara sp. and 1.0 mg kg 1 for red mullet (Mullus surmuletus). arsenic in different marine organisms: Importance in monitoring studies.
Marine Environmental Research, 58, 845–850.
MAFF (1995) has established a maximum permitted mercury level Fattorini, D., Notti, A., Di Mento, R., Cicero, A. M., Gabellini, M., Russo, A., et al.
of 0.5 mg kg 1 in fish. In a recent study of fish species from the (2008). Seasonal, spatial and inter-annual variations of trace metals in mussels
Black Sea in Turkey, similar mercury levels were reported and ran- from the Adriatic Sea: A regional gradient for arsenic and implications for
monitoring the impact of off-shore activities. Chemosphere, 72, 1524–1533.
ged from 0.025 to 0.078 mg kg 1 (Tuzen, 2009). In Spain, muscle Giordano, R., Arata, P., Ciaralli, L., Rinaldi, S., Giani, M., Cicero, A. M., et al. (1991).
mercury concentrations ranged from 0.14 to 0.36 mg kg 1 in red Heavy metals in mussels and fish from Italian coastal waters. Marine Pollution
mullet, 0.08 to 0.09 mg kg 1 in anchovy, 0.12 to 0.29 mg kg 1 in Bulletin, 22(1), 10–14.
Gümgüm, B., Unlü, E., Tez, Z., & Gülsün, Z. (1994). Heavy metal pollution in water,
hake, and 0.06 to 0.15 mg kg 1 in mackerel (Falcó et al., 2006). sediment and fish from the Tigris River in Turkey. Chemosphere, 29(1), 111–116.
In the southern Atlantic coast of Spain, mercury concentration in Güven, K., Ozbay, C., Unlü, E., & Satar, A. (1999). Acute lethal toxicity and
three fish species ranged from 0.01 to 0.023 mg kg 1 (Usero accumulation of copper in Gammarus pulex (L.) (Amphipoda). Turkish Journal of
Biology, 23, 510–521.
et al., 2003). Higher levels of mercury (0.39 mg kg 1) were found
Has-Schon, E., Bogut, I., & Strelec, I. (2006). Heavy metal profile in five fish species
in hake of the Adriatic Sea (Storelli, Storelli, Giacominelli-Stuffler, included in human diet, domiciled in the end flow of River Neretva (Croatia).
& Marcotrigiano, 2005) and black scabbard fish (0.90 mg kg 1) Archives of Environmental Contamination and Toxicology, 50, 545–551.
caught in Portugal (Afonso, Lourenc, Abreu Dias, Nunes, & Castro, Joint FAO/WHO (1999). Expert committee on food additives. Summary and
conclusions, 53rd meeting, Rome, 1–10 June.
2007). Kalay, M., Ay, O., & Canlı, M. (1999). Heavy metal concentrations in fish tissues from
In conclusion, statistically significant differences were observed the Northeast Mediterranean Sea. Bulletin Environmental Contamination and
in mean metal values obtained for fish species from the Adriatic Toxicology, 63, 673–681.
Kapaj, S., Peterson, H., Liber, K., & Bhattacharya, P. (2006). Human health effects
Sea. The results presented on metal contents in the examined spe- from chronic arsenic poisoning – A review. Journal of Environmental Science and
cies gives an indication of the environmental conditions along the Health Part A, 42, 2399–2428.
eastern coast of the Adriatic Sea. Accordingly, the concentrations of Keskin, Y., Baskaya, R., Ozyaral, O., Yurdun, T., Luleci, N. E., & Hayran, O. (2007).
Cadmium, lead, mercury and copper in fish from the Marmara Sea, Turkey.
Cd, Cu, Hg and Pb obtained were far below the established values Bulletin of Environmental Contamination and Toxicology, 78, 258–261.
by the European Community regulations. However, arsenic levels Kljaković Gašparić, Z., Zvonarić, T., Vrgoč, N., Odžak, N., & Barić, A. (2002). Cadmium
found in red mullet were higher than the recommended legal lim- and lead in selected tissues of two commercially important fish species from
the Adriatic Sea. Water Research, 36, 5023–5028.
its for human consumption and may present a human health issue. Lewis, M. A., Scott, G. I., Bearden, D. W., Quarles, R. L., Moore, J., Strozier, E. D., et al.
(2002). Fish tissue quality in near-coastal areas of the Gulf of Mexico receiving
Acknowledgements point source discharges. The Science of the Total Environment, 284(1–3),
249–261.
MAFF (1995). Monitoring and surveillance of non-radioactive contaminants in the
The authors are thankful to all those who contributed to the aquatic environment and activities regulating the disposal of wastes at sea,
study. Special thanks go to Marijana Fluka and Mirjana Hren for 1993. Aquatic Environment Monitoring Report No. 44. Directorate of Fisheries
Research, Lowestoft.
sample preparation. This study was supported with a grant from
Orescanin, V., Lovrencic, I., Mikelic, L., Barisic, D., Matasin, Z., Lulic, S., et al. (2006).
the Ministry of Agriculture, Fisheries and Rural Development Vet- Biomonitoring of heavy metals and arsenic on the east coast of the Middle
erinary Directorate, Croatia. Adriatic Sea using Mytilus galloprovincialis. Nuclear Instruments and Methods in
Physics Research B, 245, 495–500.
Shah, A. Q., Kazi, T. G., Arain, M. B., Jamali, M. K., Afridi, H. I., Jalbani, N., et al. (2009).
References Accumulation of arsenic in different fresh water fish species – potential
contribution to high arsenic intakes. Food Chemistry, 112, 520–524.
Afonso, C., Lourenc, H. M., Abreu Dias Nunes, M. L., & Castro, M. (2007). Contaminant Sepe, A., Ciaralli, L., Ciprotti, M., Giordano, R., Fumari, E., & Costantini, S. (2003).
metals in black scabbard fish (Aphanopus carbo) caught off Madeira and the Determination of cadmium, chromium, lead and vanadium in six fish species
Azores. Food Chemistry, 101, 120–125. from the Adriatic Sea. Food Additives and Contaminants, 20, 543–552.
Ordinance (2008). Ordinance setting maximum levels for certain contaminants in Storelli, M. M., Storelli, A., Giacominelli-Stuffler, R., & Marcotrigiano, G. O. (2005).
foodstuffs. Official Gazette, 154, 4198. Mercury speciation in the muscle of two commercially important fish, hake
Benoff, S., Jacop, A., & Hurley, I. R. (2000). Male infertility and environmental (Merluccius merluccius) and striped mullet (Mullus barbatus) from the
exposure to lead and cadmium. Human Reproduction Update, 6, 107–121. Mediterranean Sea: Estimated weekly intake. Food Chemistry, 89, 295–300.
Bryan, G., & Langston, W. J. (1992). Bioavailability, accumulation and effects of Storelli, M. M., Barone, G., Garofalo & Marcotrigiano, G. O. (2007). Metals and
heavy metals in sediments with special reference to United Kingdom estuaries: organochlorine compounds in eel (Anguilla anguilla) from the Lesina lagoon,
A review. Environmental Pollution, 76, 89–131. Adriatic Sea (Italy). Food Chemistry, 100, 1337–1341.
1010 N. Bilandžić et al. / Food Chemistry 124 (2011) 1005–1010

Topcuoglu, S., Kırbasoglu, C., & Güngor, N. (2002). Heavy metals in organisms and Uluozlu, O. D., Tuzen, M., Mendil, D., & Soylak, M. (2007). Trace metal content in
sediments from Turkish Coast of the Black Sea. Environmental International, nine species of fish from the Black and Aegean Seas, Turkey. Food Chemistry, 104,
1069, 1–8. 835–840.
Türkmen, A., Türkmen, M., Tepe, Y., & Akyurt, I. (2005). Heavy metals in three Usero, J., Izquierdo, C., Morillo, J., & Gracia, I. (2003). Heavy metals in fish (Solea
commercially valuable fish species from Iskenderun Bay, Northern East vulgaris, Anguilla anguilla and Liza aurata) from salt marshes on the southern
Mediterranean Sea, Turkey. Food Chemistry, 91, 167–172. Atlantic coast of Spain. Environment International, 29, 949–956.
Türkmen, M., Türkmen, A., Tepe, Y., Töre, Y., & Ates, A. (2009). Determination of Valette-Silver, N. J., Riedel, G. F., Crecelius, E. A., Windom, H., Smith, R. G., & Dolvin,
metals in fish species from Aegean and Mediterranean Seas. Food Chemistry, S. S. (1999). Elevated arsenic concentrations in bivalves from the southeast
113, 233–237. coasts of the USA. Marine Environmental Research, 48, 311–333.
Tüzen, M. (2003). Determination of heavy metals in fish samples of the middle WHO (1996). Guidelines for drinking water quality (2nd ed.). Chemical aspects.
Black Sea (Turkey) by graphite furnace atomic absorption spectrometry. Food Geneva: WHO. <http://www.who.int/water_sanitation_health/dwq/gdwq2v1/
Chemistry, 27, 521–526. en/> Accessed 10.05.10.
Tuzen, M., & Soylak, M. (2007). Determination of trace metals in canned fish Yilmaz, F., Ozdemir, N., Demirak, A., & Tuna, A. L. (2007). Heavy metal levels in two
marketed in Turkey. Food Chemistry, 101, 1378–1382. fish species Leuciscus cephalus and Lepomis gibbosus. Food Chemistry, 100,
Tuzen, M. (2009). Toxic and essential trace elemental contents in fish species 830–835.
from the Black Sea, Turkey. Food and Chemical Toxicology, 47, 1785– Zhang, I., & Wong, M. H. (2007). Environmental mercury contamination in China:
1790. Sources and impacts. Environmental International, 33, 108–121.