Environ. Sci. Technol. 2001, 35, 4163-4169
Chlorinated Cyclodiene Pesticide Residues in Blue Mussel, Crab, and Fish in the Gulf of Gdan´sk, Baltic Sea JERZY FALANDYSZ,* LIDIA STRANDBERG, TOMASZ PUZYN, AND MAGDALENA GUCIA Department of Environmental Chemistry & Ecotoxicology, University of Gdan ´ sk, 18 Sobieskiego Str., PL 80-952 Gdan ´ sk, Poland CHRISTOFFER RAPPE Environmental Chemistry, Department of Chemistry, Umeå University, S-901 87 Umeå, Sweden
Chlordane components (CHLs) and their metabolites (heptachlor, cis-heptachlor epoxide, U82, MC4, transchlordane, MC5, cis-chlordane, MC7, oxychlordane, MC6,and trans- and cis-nonachlor) and aldrin, dieldrin, endrin, isodrin, endosulfan 1, endosulfan 2, and mirex were quantified in the soft tissues of blue mussel, a whole crab, and whole fishes collected from the spatially different sites in the Gulf of Gdan´ sk. Six to twelve chlordane compounds and metabolites and dieldrin were detected in all organisms examined while aldrin, endrin, isodrin, endosulfans 1 and 2, and mirex were not found above the detection limit of the method. The lipid weight based concentrations in Baltic biota were relatively small and ranged from 12 to 150 and 7.677 ng/g, while between 0.16 and 6.8 and 0.10-6.6 ng/g in fresh tissue, respectively. The profile (%) of chlordane compounds was very similar between various fish species with trans-nonachlor (28 ( 17), cis-chlordane (23 ( 18), oxychlordane (13 ( 7), and heptachlor epoxide (11 ( 5) as major constituents and was totally different in crab with oxychlordne as the most dominating (>65%) compound. Blue mussel, lamprey, and three-spined stickleback exhibited a smallest ability to metabolize CHLs, and such fishes as cod, lesser sand-eel, sand-eel, pikeperch, perch, round goby, flounder, and herring showed a slightly better ability, while crab was able to effectively metabolize most of CHL compounds except trans-nonachlor. A value of the quotient of the trans-nonachlor to cis-chlordane concentrations (N/C quotient) was 1.0 in blue mussel, 3.1 in crab, and between 0.9 and 1.8 in fish. Both the small concentrations of CHLs in all organisms and the values of N/C quotients close to 1 imply on a long-range aerial transport through movement of the air masses from the remote regions of the northern hemisphere as a main source of this pesticide in the Gulf of Gdan´ sk. The interdependences between the CHL profiles for various fish species and between different sampling sites were examined using the principal component analysis (PCA) method. Applying the PCA model the first four significant components explained 90% 10.1021/es010059l CCC: $20.00 Published on Web 10/03/2001
2001 American Chemical Society
(43% + 23% + 15% + 8%) of the total variance in the data matrix.
Introduction The chlorinated cyclodiene pesticides were produced in large quantities after 1940s (1-3). These compounds were used mainly to control agricultural pests, insect born diseases, and for termite control. Technical chlordane is composed of more than 147 constituents of mainly C9 to C15 skeleton (1, 4), and transchlordane (24 ( 2%), cis-chlordane (19 ( 3%), heptachlor (10 ( 2%), trans-nonachlor (7 ( 3%), and cis-nonachlor (7 ( 1%) are major constituents (5), while MC4, MC5, MC6, MC7, and U82 are minor (1, 6). Heptachlor, a constituent of technical chlordane (about 10%) with a highest insecticidal activity, was also synthesized (65-72% of active agent) and sold separately as insecticide and fungicide (2, 7). Technical grade formulations of mirex consist of 95.19% mirex and by side products and including keptone (3). Many persistent, lipophilic, and toxic organochlorine compounds contaminating the environment readily bioaccumulate and biomagnify through the aquatic food chain and can be found in seafood from different parts of the world (8-10). Chlorinated cyclodiene pesticides are also highly toxic, persistent, and tend to bioaccumulate and biomagnify in the trophic webs. For example CHLs is highly persistent in soil (11) and in sediments (t1/2 ) 11-17 years) (12). Because concern over cancer risk or other toxic effects observed in man and wildlife, persistency in the environment and accumulation in biota the use of chlorinated cyclodine pesticides in many countries was restricted to combat ants and termites in soil and for protection of plants not used for food production or was prohibited (13, 14). Very limited information is available on the rates or total amount of various chlorinated cyclodiene pesticides used in the past around the Baltic Sea, and also relatively scare are data on their occurrence, concentrations, fate, and possible effects in this sea (15-21). Those compounds such as many other semivolatile and environmentally persistent organohalogenated substances are less or more prone for atmospheric transportation and subsequent deposition on the earth surface (22). Filter feeding blue mussel has very limited capacity to metabolize lipophilic organochlorine pollutants and is widely used as an indicator organism in marine pollution monitoring programs (23-25). Fish due to low activity of the monooxygenase enzymes poses very limited ability to metabolize persistent organohalogenated compound contaminating the environment and including CHLs (26, 27). The number of data published on occurrence, composition, sources of origin, and effects of chlorinated cyclodiene pesticides in Baltic fish and another biota is much less than for many other groups of persistent organochlorine compounds (28). In this study we compare concentrations and profile of the residues of chlorinated cyclodiene pesticides in various biota and including a representatives of molluscs (blue mussel), crustaceans (crab), and fishes from the southern region of the Baltic Sea. Furthermore, the objectives were to assess and explain possible origins of, and interdependences among, CHLs concentrations and profiles in biota from the Gulf of Gdan ´ sk using principal component analysis (PCA) method. * Corresponding author phone: +48 58 3450372; fax: +48 58 3410357; e-mail:
[email protected]. VOL. 35, NO. 21, 2001 / ENVIRONMENTAL SCIENCE & TECHNOLOGY
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FIGURE 1. Localization of the sampling sites.
Materials and Methods Collection of Samples. Blue mussel, crab, and fishes were collected in the Gulf of Gdan ´ sk in 1992. Blue mussel (Mytilus trossulus), crab (Carcinus means), cod (Gadus morhua), herring (Clupea harrengus), pikeperch (Stizostedion lucioperca), perch (Perca fluviatilis), round goby (Neogobius melanostomus), greater sand-eel (Hyperoplus lanceolatus), lesser sand-eel (Amodytes tobianus), lamprey (Lampetra fluviatilis), flounder (Platychthis flesus), and three-spined stickleback (Gasterosteus aculeatus) were collected at the sites shown in Figure 1. The data on the total number of the individuals investigated together with their lipids content and the residue concentrations based on a fresh and lipid weight are given in Table 1. Blue mussels were taken by grab sampler, while crab and fishes were netted using a bottom trap or gill net. Except of blue mussel all other organisms were deep frozen (-20 °C) directly after capture and kept in a such condition in clean polyethylene bags until chemical analysis. Blue mussel before dissection was kept for 24 h in 4 °C in water collected at the place of sampling. Pooled samples, containing from 3 to 30 whole crab, fishes, and soft tissue of 350 individual mussels, were subjected to chemical analysis. Chemical Analyses. The analytical method used for determination of chlorinated cyclodiene pesticides is a part of a multiresidue procedure allowing the simultaneous determination of many organochlorines and polynuclear aromatic hydrocarbons (29). After homogenization of the sample (soft part of blue mussels and whole crab and fishes; 69-380 g) with anhydrous sodium sulfate (1:7; baked at 550 °C for 2 days), a powdered mixture was packed into a wide bore open glass column, spiked with an internal standard (13C12-p,p’-DDT and dieldrin), and extracted with 500 mL of a mixture of acetone and n-hexane (2.5:1) followed by 500 mL of n-hexane and diethyl ether (9:1), to obtain a fat extract. The solvents were carefully evaporated on a water bath under vacuum pressure using a rotary evaporator. Pure ethanol was then added to remove azeotropically coextracted water also under vacuum and using the rotary evaporator. Bulk lipid removal was performed by means of the polyethylene film dialysis method (29). After dissolving the extracted lipids in cyclopentane, dialysis through the polymeric membrane was accomplished by changing the dialysate after 24, 48, and 72 h. The three-dialysate fractions, containing normally between 1 and 10% of the original lipids, depending on sample size and matrix type, were combined and concentrated to a few milliliters using a rotary evaporator. The extract was split into two parts, of which 10% was used for analysis 4164
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of cyclodienes and some other organochlorine pesticides, and bulk of polychlorinated biphenyls (PCBs), while 90% was used for the analysis of planar organochlorines, which are not reported here. The remaining lipid was removed on a Florisil column (35 cm length), and the analyte was fractionated using n-hexane (28 mL; fraction 1), methylene chloride, and n-hexane (15:85 v/v; 38 mL, fraction 2), methylene chloride and n-hexane (50:50 v/v; 56 mL, fraction 3), and methanol (66 mL; fraction 4). The cyclodienes were eluted in fractions 1 and 2, which were combined, and dieldrin in fraction 3, and the solvent was evaporated in room temperature with tetradecane added as a keeper. Before injection of the analytes to a capillary column gas chromatography/low resolution mass spectrometer system (HRGC/ LRMS) a recovery standard of (13C12-2,2’,4,5,5’-pentachlorobiphenyl; PCB no. 101) was added. Procedural blanks were performed with every set of the real samples analyzed, which only contained minor residues of PCBs and hexachlorobenzene and were well below 10% of any calculated value. The recoveries were generally between 60 and 120%, and the results were corrected for recovery values (29). Detection limit of the method (ng/g lipid) was 0.04 for heptachlor epoxide and oxychlordane, 0.05 for cis- and trans-chlordane, trans-nonachlor, MC4, MC5, MC6, MC7, and U82, 0.06 for cis-nonachlor and aldrin, 0.07 for heptachlor, dieldrin, and mirex, and 0.6 for isodrin. Statistical data analysis was performed using Principal Component Analysis (PCA) method by means of Statistica 5.0 software package. In this method, the data matrix is decomposed into a loading matrix and a score matrix following the equation A
xik ) xj k +
∑t
ia
‚pak + eik
a)1
where xik is the element of the data matrix (index i is used for samples, and index k for compounds), xj k is the mean concentration of each compound, tia is the location of the object i along the ath principal component, and pak is the value of loading, which describing how much the compound k contributes to the ath principal component. PCA method creates new orthogonal variables called principal components or PCs. They are linear combinations of redounding columns from the data matrix. The first extracted PC explains the main variation in the data, the second one describes the next largest part of total variance and so on. Comparing localization of loadings (samples) and
TABLE 1. Concentrations of Cyclodiene Pesticides in Soft Tissue of Blue Mussel, a Whole Crab, and a Whole Fish from the Gulf of Gdan´ sk matrix, ng/g fresh wt blue mussel
chlordane no.
lipids(%)
cis-
trans-
2 (700)
1.5 1.3-1.7 1.3 9.0 3.4 4.4 5.6
0.05 0.04-0.06 0.02 1.3 0.11 0.18 0.25 0.14-0.35 0.15 0.15 0.08 1.8 0.30-3.2 0.20 ( 0.04 0.15-0.24 0.44 ( 0.27 0.24-0.82
0.02 0.02-0.03 0.01 0.21 0.02 0.04 0.06 0.04-0.08 0.03 0.04 0.02 0.31 0.04-0.59 0.06 ( 0.01 0.05-0.07 0.07 ( 0.03 0.04-0.12
crab herring cod pikeperch perch
1 (3) 1 (3) 1 (3) 1 (3) 2 (16)
round goby sand-eel lesser sand-eel lamprey
1 (6) 1 (20) 1 (20) 2 (6)
4.8 5.7 5.5 15
flounder
3 (15)
4.6
stickleback
4 (120)
2.5
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matrix, ng/g lipidsa blue mussel
2 (700) 1 (3) 1 (3) 1 (3) 1 (3) 2 (16)
round goby sand-eel lesser sand-eel lamprey
1 (6) 1 (20) 1 (20) 2 (6)
flounder
3 (15)
stickleback
4 (120)
lipids(%)
cis-
trans-
3.2 2.8-3.6 1.9 14 3.1 4.0 4.5 2.4-6.6 3.2 2.5 1.5 9.6 4.8-14 4.3 ( 1.0 3.2-5.1 18 ( 11 8.9-34
1.7 1.6-1.8 0.49 2.3 0.71 0.91 1.0 0.60-1.5 0.55 0.75 0.39 1.6 0.62-2.6 1.4 ( 0.2 1.1-1.5 2.8 ( 1.4 1.6-1.7
trans-
ND
MC4
0.05 0.04-0.06 ND 0.08 0.33 1.1 0.05 0.15 0.09 0.24 0.12 0.43 0.08-0.16 0.28-0.58 0.08 0.25 0.07 0.20 0.03 0.09 0.61 2.1 0.14-1.1 0.55-3.7 0.06 ( 0.02 0.23 ( 0.08 0.04-0.08 0.15-0.31 0.22 ( 0.13 0.73 ( 0.34 0.13-0.41 0.48-1.2
chlordane no.
crab herring cod pikeperch perch
a
nonachlor
cis-
MC5
ND
MC6
0.03 0.02-0.04 ND 0.01 0.05 0.33 0.003 0.04 0.01 0.06 0.01 0.07 0.01-0.01 0.06-0.08 0.01 0.05 0.01 0.04 0.004 0.03 0.08 0.54 0.01-0.16 0.07-1.0 0.01 ( 0.01 0.05 ( 0.01 0.003-0.01 0.04-0.06 0.02 ( 0.01 0.15 ( 0.08 0.01-0.03 0.08-0.27
MC7
U82
ND
ND
ND-0.02 0.01 ND 0.15 0.07 0.02 0.01 0.02 0.18 0.07 0.01 0.04-0.09 0.01-0.01 0.03 0.02 0.03 0.01 0.01 0.005 0.30 0.08 0.08-0.52 0.01-0.15 0.02 ( 0.01 0.01 ( 0.01 0.01-0.03 0.005-0.01 0.09 ( 0.05 0.01 ( 0.01 0.05-0.15 0.005-0.02
oxychlordane ND
heptachlorepoxide
0.02 0.02-0.03 ND 0.36 0.03 0.04 0.54 0.35 0.01 0.08 0.14 0.01 0.11 0.15 0.01 0.18 0.16 0.01-0.01 0.17-0.20 0.16-0.17 0.01 0.09 0.09 0.01 0.10 0.09 0.004 0.06 0.06 0.08 0.83 0.03 0.01-0.15 0.20-1.5 0.01-0.05 0.01 ( 0.01 0.09 ( 0.02 0.10 ( 0.01 0.004-0.01 0.07-0.11 0.09-0.12 0.02 ( 0.01 0.22 ( 0.12 0.09 ( 0.01 0.01-0.03 0.11-0.37 0.08-0.11
CHLs
dieldrin
0.18 0.16-0.0.20 0.52 4.4 0.63 0.92 1.4 1.0-1.8 0.80 0.75 0.40 6.8 1.4-12 0.83 ( 0.21 0.62-1.0 2.1 ( 1.1 1.3-3.5
0.21 0.18-0.24 0.10 6.6 1.7 0.92 1.9 1.6-2.2 0.96 1.4 0.84 3.8 1.1-6.5 0.83 ( 0.21 0.62-1.0 1.2 ( 0.5 0.92-1.9
nonachlor
cisND ND 3.6 1.6 1.9 2.3 1.4-3.1 1.7 1.2 0.49 3.5 2.2-4.8 1.3 ( 0.4 0.86-1.7 9.1 ( 5.4 5.3-17
trans3.3 2.8-3.7 6.3 12 4.5 5.4 7.9 4.7-11 5.2 3.6 1.7 13 9.0-16 5.0 ( 1.7 3.1-6.4 30 ( 14 19-49
MC4
MC5
ND 0.54 0.09 0.12 0.16 0.09-0.22 0.11 0.11 0.07 0.42 0.13-0.72 0.12 ( 0.05 0.06-0.15 0.75 ( 0.56 0.19-1.2
1.7 1.2-2.2 0.92 3.7 1.0 1.4 1.3 1.0-1.5 1.1 0.76 0.56 2.8 1.1-4.5 1.1 ( 0.2 0.79-1.3 6.2 ( 3.4 2.9-11
ND
MC6
MC7
0.76 1.7 0.56 0.46 1.3 0.72-1.8 0.53 0.46 0.19 1.8 1.2-2.3 0.46 ( 0.18 0.26-0.59 3.6 ( 2.0 2.0-6.3
ND-1.4 ND 0.73 0.18 0.55 0.20 0.12-0.27 0.33 0.11 0.08 0.39 0.12-0.66 0.16 ( 0.05 0.10-0.20 0.42 ( 0.22 0.17-0.70
ND
Heptachlor (
