Organochlorines in benthic invertebrates and sediments from the Dutch Wadden Sea; identification of individual PCB components

Netherlands Journal of Sea Research 17 (1). 19-38 (1983)

ORGANOCHLORINES IN BENTHIC INVERTEBRATES AND SEDIMENTS FROM THE DUTCH WADDEN SEA; IDENTIFICATION OF INDIVIDUAL PCB COMPONENTS by J.C. DUINKER*,

M.T.J. HILLEBRAND

andJ.P. BOON**

Netherlands Institute for Sea Research, P.O. Box 59, 1790 AB Den Burg, Texel, The Netherlands

CONTENTS 1. Introduction . . . . . . . . . . . . . . . . . . . . . 2. Material and Methods . . . . . . . . . . . . . . . . . . . . . . 2.1. Collection and composition of samples . . . . . . . . . . . . 2.2. Analytical procedure . . . . . . . . . . . . . . . . . . . . 3. Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1. PCB patterns . . . . . . . . . . . . . . . . . . . . . . . 3.1.1. Macoma balthica and Arenicola marina . . . . . . . . . . 3.1.2. Crangon crangon . . . . . . . . . . . . . . . . . . . 3.1.3. Sediment 3.1.4. Comparison with Clophen mixtures . . . . . . . . . . 3.2. Pesticides . . . . . . . . . . . . . . . . . . . . . . . . . 3.3. Geographical trends . . . . . . . . . . . . . . . . . . . . 4. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . 5. Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . 6. References . . . . . . . . . . . . . . . . . . . . . . . . . . .

19 20 20 21 22 22 25 25 28 28 28 29 31 36 36

i. I N T R O D U C T I O N The presence of high tissue concentrations of certain organochlorines in o r g a n i s m s f r o m t h e D u t c h W a d d e n S e a h a s b e e n a s s o c i a t e d w i t h a s t r o n g d e c l i n e o f t h e p o p u l a t i o n o f f i s h - e a t i n g b i r d s (KOEMAN & VAN GENDEREN, 1966, 1972; SWENNEN, 1972) a n d m a r i n e m a m m a l s (REIJNDERS, 1980). I n i n v e r t e b r a t e s , o r g a n o c h l o r i n e c o m p o u n d s c a n be taken up from water, seston, sediment and food. Elimination can o c c u r t h r o u g h faeces, e x c r e t a a n d e q u i l i b r i u m p a r t i t i o n i n g w i t h a m b i e n t w a t e r (HAMELINK, WAYBRANT • BALL, 1971; COURTNEY & DENTON, 1976; ERNST, GOERKE & WEBER, 1977; GOERKE & ERNST, 1977, 1980; FOWLER et al., 1978; ELDER, FOWLER & POLIKARPOV, 1979; COURTNEY & LANGSTON, 1978; M c L E E S E , METCALFE ~L PEZZACK, * Present address: Institut ffir Meereskunde an der Universitfit Kiel, Abteilung Meereschemie, Dfisternbrookerweg 20, D 2300 Kiel, F.R.G. ** To whom correspondence should be directed.

20

j.c.

DUINKER

ET AL.

1980). Metabolism may also be important (ERNST, GOERKE & W E B E R , 1977; GOERKE &: ERNST, 1980). Adult benthic organisms present in the Wadden Sea may migrate passively within this limited a r e a (VERWEY, 1978). Brown shrimps enter a particular area of the Wadden Sea in the spring and remain there during the whole summer season (BoDDEKE, t975, 1976). Therefore, the concentration of organochlorines present in the tissues of the species collected there for this study will reflect the contamination of that environment, however with some restrictions. For example, the degree of bio-accumulation of compounds depends also on the rates of uptake and elimination. The latter rate can be enhanced by biotransformation. The organochlorines dissolved in the waters of the western Dutch Wadden Sea originate mainly from the river Rhine (DuINKER & HILLEBRAND, 1979). Its influence decreases toward the eastern part of the Wadden Sea. This partly explains the lower concentrations of certain organochlorines in organisms of the German Wadden Sea than in those organisms of the western (Dutch) Wadden Sea (EDER et al., 1981). The aim of this study is (1) to establish whether benthic invertebrates from different parts of the Dutch Wadden Sea possess different tissue levels of organochlorine compounds, (2) to establish the presence of any correlation between animal tissue concentrations with size or age as well as with the levels of organochlorines in bottom sediments, (3) to identify individual polychlorobiphenyls (PCB) components. The compositions of PCB in invertebrate species and in sediments will be compared with those in technical ff)rmulations. Acknowledgements.--The authors are gratetul to Dr,J.M. Everaarts for his critical reading of the manuscript and his useful comments, and to the crews of the RVs Eider, Griend and Ephyra for their valuable cooperation during sampling. We also wish to thank Mr F. Eijgenraam for designing a computer program that enabled us to plot concentrations of individual PCB components and pesticides. 2. M A T E R I A L 2 1. C O L L E C T I O N

AND

METHOI)S

AND C O M P O S I T I O N

OF S A M P L E S

Samples of organisms and sediments were obtained in .June 1979 at 5 stations (Fig. 1). The bivalve Macoma balthica (L.) and the polychaete arenicola marina (L.) (lugworm) were collected directly from the sediment of tidal fiats by means of stainless steel tweezers. Samples of

ORGANOCHLORINES 3° 54*

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21

INVERTEBRATES 8* ,

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Fig. 1. T h e 5 stations in the D u t c h W a d d e n Sea.

Crangon crangon (L.) (brown shrimp) were obtained by beam-trawling in the channels (stations 3 and 4 only). The animals were washed with sea water and wrapped in aluminium foil (pre-cleaned with hexane) and deep frozen at - 2 0 ° C within 2 hours. Sediment samples were stored at - 2 0 ° C in glass jars covered with aluminium foil. Grain-size distributions were determined by dry sieving. 2. 2. A N A L Y T I C A L

PROCEDURE

The animal samples, each containing at least 10 individuals, were ground in a Prolabo mechanical shaker. Of Macoma only the soft parts were taken• A mixture of 3 to 4 g homogenate and about 40 g anhydrous sodium sulphate was Soxhlet-extracted with n-pentane (nanograde ®, Malinckrodt) during 8 hours. An aliquot of (not more than 70 mg) lipid material was subjected to clean-up with A1203 and was fractioned with SiO 2 according to the procedure designed by H O L D E N • M A R S D E N (1969) and improved by D U I N K E R • H I L L E B R A N D (1978).

22

J.C.

D U 1 N K E R ET AL.

The compound p,p'-DDE co-eluted with some PCB components. Therefore it was quantified as the difference between the (packed column G L C - E C D ) chromatograms of the (PCB containing) first silica fraction before and after oxidation with chromic acid (Merck) ( D u I N K E R & H1LLEBRAND, 1979). This chromic acid treated first fraction and the second fraction were analysed by temperature-programmed capillary gas liquid chromatography (GLC) on a Hewlett Packard 5880A gaschromatograph equipped with a 63Ni electron capture detector (63Ni-ECD). The first silica fractions of samples of Macoma balthica, Arenicola marina and sediments, and all second silica fractions were analysed on a wall-coated open tubular ( W C O T ) 25 m x 0.22 mm CP-Sil 5 column: carrier gas He 130 kPa; make-up gas N~ 30 ml.min-1; septum purge He 5 ml- min-1; injector purge He 20 m l - m i n < ; injector 230°C, detector 320°C; temperature program: isothermal phases at 60°C (2 min), 180°C (15 min), 210°C (5 min) and 250°C (10 min) with intermediate temperature increase rates of 25 °, 4 ° and 4°C . min -1 respectively. Injection 1 /xl splitiess with a Hewlett Packard autosampler, type 7672A. Samples of the first silica fractions of Crangon crangon and one sample of Macoma were analysed on a 50 m x 0.23 mm SE-54 column. Chromatographic conditions were only slightly different in the following aspects: pressure carrier gas 170 kPa; injector 250°C; isothermal phases at 60°C (2 min), 180°C (8 min), 220°C (5 rain) and 250°C (25 min) with intermediate temperature increase rates of 10 °, 4 ° and 4°C • min 1 respectively. The analyses of the Macoma sample gave almost identical results on both columns. All glassware used was heated at 350°C for 12 h and cleaned with hexane (nanograde ®, Mallinckrodt) betore use. 3. R E S U I , T S 3 1. PCB P A T T E R N S

With the use of packed G L C colunms, PCB analyses are semiquantitative in terms of technical formulation equivalents as e.g. Clophens, Aroctors (DuINKER et al., 1980). The greatly improved separation offered by temperature programmed capillary G L C allows identification and quantification of several individual components. However, neither is separation of all components on one column perfect, nor are all components of interest available as references. Therefore detailed information is given about which PCB components were co-eluting and which were well separated (Table 1). The 209

ORGANOCHLORINES IN BENTHIC INVERTEBRATES i

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Fig. 2. PCB components and pesticides in Macoma balthica at 4 stations. For both PCB and pesticides, the concentration in n g . g-i wet weight and in # g . g-1 (pentane extractable) lipid of the component with highest concentration, are given in the upper left hand corner of each histogram. In the case of PCB, total PCB concentration on a lipid basis ( # g . g ~) is given as a third value. The histograms give proportional concentrations, of components above 10% of the component with highest concentration above the abscissa, and of components below 10% below the abscissa but on a 10 times larger scale (nd means no data). The PCB components are given in order of elution, numbered according to BALLSCHMITER & ZELL (1980); for co-eluting components quantitation is on basis of the first (upper) component given or (when + between numbers) on basis of the average response factor.

24

J.C.

,

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O R G A N O C H L O R I N E S

IN

B E N T H I C

25

I N V E R T E B R A T E S

theoretically possible PCB components are distinguished by the numbering system of BALLSCHMITER & ZELL (1980) according to I U P A C rules. 3.

1.

1.

M A C O M A

B A L T H I C A

A N D

A R E N I C O L A

M A R I N A

The chromatograms of the first silica fraction of these species were similar (Figs 2 and 3; Table 1). The dominant PCB components are 101, 149, 118, 138, 153, 180 and 187. Concentrations of 187 were probably overestimated in all samples, because of a significant peak in the chromatograms of the blank procedure. Component 118 was only available as reference component as a dilute solution. The response factor calculated was unexpectedly low and concentrations in samples may thus be overestimated. Component 105 was clearly visible as a shoulder of 132. 3.

1.2.

C R A N G O N

C R A N G O N

The PCB pattern in Crangon crangon (Fig. 4; Table 1) differed from l

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Fig. 4. Contents of PCB and pesticides in Crangon crangon at 2 stations. For explanation see Fig. 2.

26

J.C.

DUINKER

E?

AL.

%TTTrrT ,12 I 66 PC8

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Fig. 5. Contents of P C B and pesticides in sediments in n g . g-~ dry weight of the c o m p o n e n t with highest concentration and total P C B from 9 samples at 5 stations. For explanation of histograms see Fig. 2.

t h o s e of Macoma balthica and Arenicola marina in s o m e r e s p e c t s : t h e relative concentrations of components 52, 44, 95/66, 110/77 and 149

28

j.c.

DUINKER

ET

AL.

were lower (the notation " / " represents co-eluting components). On the CP-Sil 5 column, component 132 co-eluted as a shoulder of 153, while 105 was well separated from this peak. 3. I ~

SEDIMENT

In sediments, a number of di-, tri- and tetrachlorobiphenyls were found in relatively high concentrations (Fig. 5; Table 1). Partly these components (18, 31 and 37/42) were also found (at least occasionally) in the invertebrates investigated, but components 15 and 28 were found only in sediments. The components with lower contributions were below detection level in most coarse sediments. 3 1. 4

COMPARISON

WITH

CLOPHEN

MIXTURES

In the past, PCB contents have been expressed almost exclusively in terms of technical formulation equivalents, involving the comparison of packed column chromatograms of environmental samples with those of a particular formulation. In practice, only tormulations with a high degree of chlorination have been used for comparison (such as Clophens A50 and A60 or Aroclors 1254 and 1260). It is interesting therefore to compare the present capillary column chromatograms of the PCB fraction of animals and sediments with those of technical formulations, such as Clophens A30, A40, A50 and A60, in terms of individual components. The compositions of the formulations are taken from DUINKER & HILLEBRAND (1983). The chromatograms of the environmental samples could not be reproduced by any one of the single technical formulations. This was also reported for sea water extracts (DuINKER et al., 1980). The region with the strongest peaks in the chromatograms of the present samples matches the regions of strongest peaks in Clophens A50 and A60. Some of the most important components in the samples are dominant in Clophen A50 (101, 110/77, 118) while others dominate in Clophen A60 (149, 153, 138, 187, 180). Components present only in Clophens A30 and A40 were not found at all or only occasionally and in low concentrations in the invertebrates investigated. 2

PESTIC,

1 DES

Pesticides detected in the first silica fraction were HIS:B, and p,p'-DDE. The second silica fraction showed the presence of o~- and 3'-HCH, dieldrin and p , p ' - D D D . The compound j3-HCH and the cyclodienes aldrin, endrin, heptachlor and (its metabolite) hep-

ORGANOCHLORINES

IN

BENTHIC

29

INVERTEBRATES

tachlorepoxide could not be detected. A compound with the retention time of mirex could only be determined on the CP-Sil 5 column; concentrations (of mirex) on a lipid basis were 0.3 to 1.0 /zg. g-i (not shown). The presence of o , p ' - D D D and p , p ' - D D T , if occurring at all in the samples, is doubtful because of interference with large peaks of unknown components. GEOGRAPHICAL

3.3.

TRENDS

Organochlorine concentrations (Figs 2 to 4), show considerable similarity on a lipid basis in samples from the same station, irrespecw'

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Fig. 6. Contents of PCB and pesticides in 3 different age classes of Macoma balthica from station 1. For explanation of histograms see Fig. 2.

30

j.c.

DUINKER

ET

AL.

tive of a n i m a l species. T h e relation between P C B and pesticide concentrations was investigated in different age classes of Macoma balthica f r o m station 1 (Fig. 6) and station 5, a n d in different size classes of Crangon crangon f r o m station 4. C o n c e n t r a t i o n s on a lipid basis appeared to be i n d e p e n d e n t of size or age. T h e r e t o r e , these concentrations can be used to express the geographical (west-east) trend. Since P C B p a t t e r n s in a single a n i m a l species did not vary with concentration, any single P C B c o m p o n e n t m i g h t be selected for comparison. T h e s u m of" concentrations of individual P C B c o m p o n e n t s , r e p r e s e n t i n g total P C B in the sample, m a y be an even m o r e useful and accurate basis for c o m p a r i s o n of c o n t a m i n a t i o n levels in samples of the same species, o r i g i n a t i n g from different areas. T h i s was done for total P C B of the present samples (Figs 2 to 4). T h e concentrations on a lipid basis of the various c o m p o u n d s in Arenicola marina and Macoma balthica were c o m p a r e d between the s a m p l i n g stations (Fig. 7). P C B , 7 - H C H , and to a lesser degree

401 (I

20

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/

5-

4-

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2

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Fig. 7. Relative concentrations of PCB (C)), HCB (0), c~-HCH (~), y-HCH (11), dieldrin ( zx) and p,p'-DDD ( • ), on a lipid basis in Arenicola marina (a) and Macoma balthica (b) from stations 1, 3, 4 and 5 compared to those from station 2. dieldrin were highest in samples f r o m station 1. Relatively low cont a m i n a t i o n levels were found in animals f r o m stations 2, 3 and 4. T h e high values in Macoma at station 5 coincided with a tow lipid content of a n i m a l s f r o m this area; the concentrations on a wet weight basis were a m o n g the lowest f o u n d (see Fig. 2d). A p r o n o u n c e d m a x i m u m of H C B on a lipid basis (one order of m a g n i t u d e a b o v e m i n i m u m values) was f o u n d in Macoma and Arenicola

ORGANOCHLORINES IN BENTHIC INVERTEBRATES

31

from station 5 and in Crangon from station 4 (Figs 2 to 4). An increased PCB level was also found in Arenicola from station 4. No geographical trend in concentrations was found in sediments from the various stations. Over-all contents of organochlorines in sediments appeared to be determined primarily by the grain-size distribution (Fig. 5). A significant positive correlation ( p < 0 . 0 5 ) was established between the fraction of particles < 50 #m and levels of total PCB, p , p ' - D D D and p,p'-DDE. No significant correlations were found for HCB, or- and 3,-HCH, and dieldrin (Students t-test after sinus transformation of percentages). Absence of a correlation between contents of chlorinated hydrocarbons in invertebrates and sediments was also reported for a coastal area of the North Atlantic (McLEESE, RAY ~¢ METCALFE, 1979). A peak with the retention time of PCB component 26 showed a remarkable behaviour; it was relatively high in samples from stations 4 and 5, while this was not observed for other PCB components. This peak may therefore have arisen from another compound.

4. D I S C U S S I O N

The relatively high concentrations of PCB, y - H C H , and dieldrin in organisms from station 1 and a decrease in levels to minima at station 2 corresponds with the decreasing influence of the river Rhine in the Wadden Sea from west to east (ZIMMERMAN, 1976; DUINKER &: HILLEBRAND, 1979). The influence of the Ems-Dollard estuary on the adjacent area of the Dutch Wadden Sea decreases from station 5 to the west. This is particularly evident for H C B , for which the Ems-Dollard area is the most important source. Stations 2 and 3 represent the area of lowest fresh water influence in the Dutch Wadden Sea. Contamination levels in Macoma balthica and Arenicola marina from station 3 were somewhat higher than those from station 2. PCB patterns in Arenicola (a deposit feeder), and Macoma (a mixed suspension and deposit feeder) were very similar. It seems appropriate therefore to compare the PCB patterns of these species with seston and sediments, as these are both their hide and food source. The carnivorous Crangon crangon (PLAGMAN, 1939) represents another trophic level. The upper layer of sediments can be considered, however, as an important source for uptake of organochlorines by crustaceans (McLEEsE, METCALFE• PEZZACK, 1980). As aquatic animals are exposed to the ambient water (via gills and skin), water must also be considered as a significant source for uptake of organochlorines. Internal concentrations of each single PCB component in animals

32

j.c.

DUINKER

ET

AL.

depends on rates of uptake and excretion. The former factor depends on PCB concentrations in environmental compartments from which uptake takes place, i.e. water, seston, sediment and food. The dominant components in water correspond in general terms with technical formulations of low degree of chlorination, such as Clophens A30 and A40 (DuINKER et al., 1982a, 1982b), while the dominant peaks in seston and sediments correspond with dominant peaks in more heavily chlorinated mixtures, e.g. Clophens A50 and A60 (section 3.1.4; DUINKER et al., 1982a, 1982b). The relative concentration of individual components in a (theoretical) mixture, containing equal contributions of the 4 technical Clophen formulations might therefore be considered as a first, admittedly crude, approximation of the availability of individual PCB components to the invertebrates investigated. Table 1 summarises the composition of this mixture on the basis of the recent G L C - E C D and GC-MS analyses of Clophen mixtures in terms of 102 available individual components (DuINKER & HILLEBRAND, 1983). For comparison, these components are ordered in groups of similar relative concentrations. Comparing the relative concentrations of individual PCB components with this model mixture explains many differences in tissue concentrations of components with the same degree of chlorination; some components are present in higher concentrations in the environment than others, irrespective of relative persistence. Within each group of similar relative concentrations, components of higher degree of chlorination (penta- and higher) have the highest concentrations in animals. This may be due to a more rapid uptake of these components (LANGSTON, 1978) or tO a higher persistence, or both. Since metabolism of pentachlorobiphenyls and higher chlorinated biphenyls by microorganisms has not been reported and the concentrations of these components in water are extremely low, the invertebrates appear to take them up mainly from sediments and food. Generally the results are in agreement with equilibrium partitioning between body lipids and water being the dominating mechanism for determining tissue concentrations of PCB components. This is based on observations following. Although the concentrations of persistent PCB components and pesticides on a wet weight basis differ considerably between different species from the same area, the concentrations on a lipid basis are much less variable (values given in Figs 2 to 4). The concentrations on a lipid basis do not increase with size or age in samples of Macoma and Crangon, originating from the same sampling area. Since tissue concentrations in plankton appear to be determined by

ORGANOCHLORINES IN BENTHIC INVERTEBRATES

33

the same mechanism (UREY, KRICHER • BOYLAN,1976; CANTONet al., 1977; CLAYTON, PAVLOU& BREITNER, 1977; PAVLOU ~: DEXTER, 1979; YOUNG, SWADER & BINGHAM, 1980), no strong biomagnification is likely to take place in this part of the ecosystem. A comparison of the relative concentrations of PCB components in the inv.ertebrates, seston and sediment shows that components eluting after 70/80/98 (i.e. pentachlorobiphenyls and higher chlorinated biphenyls) form a similar pattern in these different compartments (some exceptions are observed in Crangon). Elimination must occur, but it does not affect the relative concentrations. Therefore it is likely that for these components the same mechanism as for the organisms, i.e. equilibrium with the ambient water also dominates in sediment and seston. STEEN, PARIS & BAUGHMAN (1978) also showed that, depending on the circumstances, sediment can act as a sink as well as a source for PCB by partitioning with the overlying water. PAL, WEBER & OVERGASH (1980) studied the velocity of leaching of PCBs from different sediment types and found that sandy l o a m > s i l t y loam > silty clay loam. WILDISH et al. (1980) found greater concentration factors for desorption than for adsorption to sediments, indicating that PCB (as Aroclor 1254) was firmly bound to the sediment. However, some desorption did occur. Compared to sediments and seston, some di-, tri-, and tetrachlorobiphenyls eluting before component 70/80/98 were present in relatively low concentrations or even missing in the species investigated. Most of these components have been found as major constituents in river water solution. The presence of a mixed function oxidase ( M F O ) system is important with respect to metabolism of PCB components in animals. The microflora of the gastro-intestinal tract might also be of importance, especially for metabolism of components with lower chlorine content. According to SUNDSTR()M, HUTZINGER& SAFE (1976), hydroxylation of PCB components appears to occur preferably at the meta (3, 5) or para (4) positions (with respect to the inter-ring bond) of the less chlorinated aromatic ring. The reactions may be affected also by the chlorine substitution pattern in the ortho (2, 6) positions. The presence of vicinal H atoms may also decrease the persistence to metabolism 0ANSSON et al., 1975; BALLSCHMITER,YELL & NEU, 1978). Therefore, different structural features of PCB components may influence the possibilities to be metabolised (Table 1). In shrimps, some of the higher chlorinated components are present in lower concentrations than in Macoma, Arenicola and sediments (101, 149), or are below detection limits (110/77). In Clophens, the contribution of 77 is small with respect to its co-eluting component 110.

34

J.C.

DUINKER

ET

TABLE

AL.

1

C o m p o s i t i o n of a theoretical P C B m i x t u r e , c o n t a i n i n g equal a m o u n t s of C l o p h e n s A30. A40, AS0 a n d A 6 0 (DuINV,ER & HILLrmRANO, 19831 used as a n estimate of the occurrence of P C B c o m p o n e n t s in t h e e n v i r o n m e n t . T h e individual c o m p o n e n t s are a r r a n g e d in 5 g r o u p s a c c o r d i n g to p e r c e n t a g e c o n t r i b u t i o n in the m i x t u r e , a n d w i t h i n g r o u p s in o r d e r of e l u t i o n f r o m the G L G c o l u m n ( I U P A C n u m b e r s a c c o r d i n g to BA~LSCnMrrER & Z~LI~, 1980); the c o n t r i b u t i o n s of co-eluting c o m p o n e n t s are e n t e r e d as t h e i r s u m . R e l a t i v e peak h e i g h t s o f Pq~B c o m p o n e n t s in each a n i m a l species a n d in s e d i m e n t s are qualitatively i n d i c a t e d by a n i n c r e a s i n g n u m b e r of plusses a n d by t h e symbols - (below detection limit) a n d ] (occasionally found). C h l o r i n e s u b s t i t u t i o n characteristics w h i c h were c o n s i d e r e d to influence persistence, are also given. N.B. peak 21/33/53 s h o w e d a slightly diffcrent r e t e n t i o n t i m e in t h e samples w h i c h shift in r e t e n t i o n t i m e m a y result f r o m the absence of o n e of the c o m p o n e n t s ; c o m p o n e n t s 118 a n d 187 are p r o b a b l y o v e r e s t i m a t e d in the sample~; in Crangon peak 153 is peak 1531132 a n d p e a k 132/105 is peak 105 because of the use of a different c o l u m n

PCB components

Relatwe peak heights Mazorna

Arenkola

Crangon

Chlorine ~ubsti~ution Sediments

Group_<0.5% 4/10 7/9 i9 24 29 46 67 88 83 200 372 397 201 196 189 194 206 209

1 i

+ ,-

0 5<~ < G r o u p - -< 1 c5~ 26 4O 86197 320 336 82 141/179 176 183 128/167 156/202 1% < G r o u p - < 2 ~ 5/8 21/33/53 49 37/42 41 61 60/71 84 99 87/90/I 16 151 187 174 177

Numb q/" Cl atoms

Numb of ortho-CI

2/2 2/2 3 3

2/2 l/l 3 2

3

1

4 4 5 5

1 3 2

?

2

•

8

4

•

8

3

• •

7

1

8

'2

•

I0

4

•

A !

++

+4 ++

+ ÷+

i

~+

+++ -*

++

I

+. +

+ + + * ~ ++

+ + . + 4 ++

+ + .

++

+ + ) + + ++

~ + ~ + ~ ++

+++

+ + ,

170

2 ~ < ( ; r o u p ~<3+;;i 15 31

4 515 5 li 5 6i7 7 7 6/6 6/8

2 2/2

2/2 31314 't 3!4 4 4 "114 5 5 51515 6 7 7 7 7

1/I 1!I!3 2 (t/2 2

I

4 2 214 4 3 2/1 1/4

• ( 1 2 8 + 1671 - ( 1 2 8 4 167) *(2021 0(156) •(202)

•(37)

1

112 2 21212 3 3

•(60)

!

'.3

3 2

@

!

l

180

I

÷ ~ +

@

2

+

3

+ +

3

1

+ * +

4/4/5

11013

+ + + + + + + +

6 IV5

+.

2 + + *

+ ÷ +

+ .

4 ÷ ÷ ~ + +

lOl

1533 1 3 2 / 1 O5

At 3,3' and 5,5'

•

i

3~
At 4,4'

I

t +

No ticinal H atoms

•(8o)

e(8I)i

2

+ ++ +

, ~ + + ~

7

3/

I

@

9

•ll05)

ORGANOCHLORINES

IN

BENTHIC

35

INVERTEBRATES

TABLE 1 (continued) PGB components

Relative peak heights Mazoma

Arenicola

Crangon

Chlorine substitution Sediments

Numb. of Clatoms

Numb. of No vicinal ortho-Cl H atoms

At 4,4'

At 3,3' and 5,5'

Group > 4 % 52 95/~

+++ + + +

+++ + + +

110/77

+ + +

+ + +

149 118 138

++++ +++++ + + + + +

+++++ +++++ +++++

++ + + + +++++ +++++

+++ + + +

4 5/4

2 3/1

+ + +

5

2

6 5 6

3 1 2

+++++ +++++ +++++

•(66)

Components 101, 110 and 149 have two features that may make them less persistent to metabolism: the presence of vicinal H atoms, and the lack of a 4,4' chlorine substitution. LEE (1981) detected considerable M F O activity in arthropods. However, Crangon was not investigated. Dominant components in the invertebrates investigated either do not possess vicinal H atoms (187) or do have a 4,4' chlorine substitution (118, 138), or both (153, 180). Neither chlorine substitution of the meta positions (i.e. 3,3', 5,5') nor an increase in the number of orthoC1 appears to increase persistence if vicinal H atoms are present and a 4,4' substitution is absent. However, these factors may increase persistence otherwise. Metabolism of components 4, 31 and 99 in Nereis virens (polychaete) was found by ERNST, GOERKE & WEBER (1977). Elimination rates (i. e. disappearance of parent components plus polar metabolites) were considerably faster for components 4 and 31 (about 3% left after 20 weeks) with respect to 99 (about 25% left after 20 weeks). Analyses of the same samples on both a packed and a capillary G L C column resulted in considerable differences. PCB concentration, calculated as sum of concentrations of individual components, are 30 to 70% below values from packed column chromatograms (as Clophen A50 equivalents). HCB levels are 60 to 90% lower. This results probably from the separation of HCB from an adjacent component, from which it was not separated on a packed column. The concentrations of some other pesticides were higher with capillary column separation, i.e. oe-HCH, 7 - H C H , dieldrin and p,p'-DDD. The single (small) peak of p , p ' - D D T in samples of organisms on the packed colu m n was separated into at least three peaks on the capillary column. However, a considerably larger peak, with the retention time of p , p ' - D D T was present in some sediment samples. Occasionally o , p ' - D D D appeared as a small peak next to a large peak of an unknown component which was probably mistaken as o,p'-DDD in the study of EDER et al. (1981). Pentachlorobenzene, often reported in our papers (e.g. DUINKER & HILLEBRAND, 1979a) co-eluted with a peak in the blank chromatogram. For these reasons it is difficult to compare the present results with previous studies, all involving packed column GLC-ECD.

36

j.c.

DUINKER

ET AL.

5. S U M M A R Y of organochlorines were d e t e r m i n e d in Macoma balthica, Arenicola marina, Crangon crangon and sediments from the D u t c h Concentrations

W a d d e n Sea. M a x i m u m concentrations of (total) P C B , " y - H C H and dieldrin were f o u n d in the western part; a p r o n o u n c e d m a x i m u m of H C B was found in the eastern part. C o n c e n t r a t i o n s of P C B , p , p ' - D D D and p , p ' - D D E in sediment were correlated with the percentage of particles < 50 #m. T h e relative c o n t r i b u t i o n of each individual c o m p o n e n t to total P C B is very similar in Macoma and Arenicola and is i n d e p e n d e n t of the c o n c e n t r a t i o n of total P C B . T h e relative concentrations of some c o m p o n e n t s with structures facilitating metabolism, were lower in Crangon. T h e patterns of pentachloro- a n d higher chlorinated biphenyls were equal in Macoma, Arenicola a n d sediments. Since these c o m p o n e n t s are not f o u n d in solution, they must be taken up from sediment and food. T h e results are in a g r e e m e n t with partitioning between b o d y lipids and water as the d o m i n a n t m e c h a n i s m d e t e r m i n i n g concentrations of persistent lipophilic c o n t a m i n a n t s in the animals. Partitioning with the ambient water m a y also determine the concentrations in the sediment.

6. R E F E R E N C E S BALLSCHMITER, K.

& M. ZELL, 1980. Analysis of polychlorinated biphenyls (PCB) by glass capillary gas chromatography. Composition of technical Aroclor and Clophen-PCB mixtures.--Z, analyt. Chem. 302: 20-31. BALLSCHMITER, K., M. ZELL • H.J. NEU, 1978. Persistence of PCB's in the ecosphere: will some PCB components "never" degrade?--Chemosphere 2: 173-176. BODDEKE,R., 1975. Autumn migration and vertical distribution of the brown shrimp CrangoncrangonL. in relation to environmental conditions. In: H. BARNES.Ninth European Marine Biology Symposium. Aberdeen University Press, Aberdeen: 483-494. - - - - , 1976. The seasonal migration of the brown shrimp Crangoncrangon.--Neth. J. Sea Res. 10 (1): 103-130. CANTON,J.H., G.J. VAN ESCH, P.A. GREVE ~¢ A . B . A . M . VAN I-IELLEMOND, 1977. Acc u m u l a t i o n and elimination of ~-hexachlorocyelohexane (~-HCH) by the marine algae Chlarnydomonasand Dunaliella.--Water Res. 11: 111-115. CLA'CTON,J.R., S.P. PAVLOV& N.F. BREITNER, 1977. Polychlorinated biphenyls in coastal marine zooplankton: bioaccunmlation by equilibrium partitioning.-Environ. Sci. Technol. 11: 676-682. COURTNEV,W.A.M. & G.R. DE~TON, 1976. Persistence of polyehlorinated biphenyls in the hard clam (Mercenariamercenaria)and the effect upon the distribution of these pollutants in the estuarine environment.--Environ. Pollut. 10: 55-63. COU~TNZV,W.A.M. & W.J. I~AN(;STON,1978. Uptake ofpolychlOrinated biphenyl (A

O R G A N O C H L O R I N E S IN B E N T H I C I N V E R T E B R A T E S

37

1254) from sediment and from seawater in two intertidal polychaetes.--Environ. Pollut. 15: 303-309. DUINKER,J.C. & M.T.J. HILLEBRAND,1978. Minimizing blank values in chlorinated hydrocarbon analysis.--J. Chromat. 150: 195-199. - - - - , 1979a. Behaviour of PCB, pentachlorobenzene, hexachlorobenzene, c~-HCH, 3~-HCH,/3-HCH, dieldrin, endrin and p,p'-DDD in the Rhine-Meuse estuary and the adjacent coastal area.--Neth. J. Sea Res. 13 (2): 256-281. - - - - , 1979b. Mobilization of organochlorines from female lipid tissue and transplacental transfer to fetus in a harbour porpoise (Phocoena phocoena) in a contaminated area.--Bull, env. contam. Toxicol. (U.S.) 23: 728-732. - - - - , 1983. Characterization of PCB components in Clophen formulations by capillary GC-MS and GC-ECD techniques.--Environ. Sci. Technol. 17: 449-456. DUINKER,J.C., M.T.J. HXLLEnRAND,R.F. NOLTING& S. WELLERSHAUS,1982a. The river Elbe: processes affecting the behaviour of metals and organochlorines during estuarine mixing.--Neth, j . Sea Res. 15 (2): 141-169. - - - - , 1982b. The river Weser: processes affecting the behaviour of metals and organochlorines during estuarine mixing.--Neth. J. Sea Res. 15 (2): 170-195. DtJINr:Ea, J.C., M.T.J. HILLEBRAND,K.H. PALMORK& S. WILHELMSEN,1980. An evaluation of existing methods for quantitation of polychlorinated biphenyls in environmental samples and suggestions for an improved method based on measurements of individual components.--Bull, env. contam. Toxicol. (U.S.) 25: 956-964. EDER, G., W. ERNST, H. GOERKE, J.C. DUINKER & M.T.J. HILLEBRAND, 1981. Organochlorine residues analysed in invertebrates of the Dutch Wadden Sea by two methods.--Neth. J. Sea Res. 15 (1): 78-87. ELDER, D.L., S.W. FOWLER& G.G. POLIKARPOV, 1979. Remobilization of sediment associated PCB by the worm Nereis diversicolor.--Bull, env. contain. Toxicol. (U.S.) 21 (4): 448-452. ERNST, W., H. GOERKE & K. WEBEr, 1977. Fate of ~4C labelled di-, tri-, and pentachlorobiphenyl in the marine annelid Nereis virens. II: Degradation and faecal elimination.--Chemosphere 9: 559-568. FOWLER, S.W., G.G. POLIKARPOV,D.L. ELDER, P. PARSl &J.-P. VILLENEUVE, 1978. Polychlorinated biphenyls: accumulation from contaminated sediments and water by the polychaete Nereis virens.--Mar. Biol. 48: 303-309. GOERKE, H. & W. ERNST, 1977. Fate of ~4C labelled di-, tri-, and pentachlorobiphenyl in the marine annelid Nerds virens. I: Accumulation and sedimentation after oral administration.--Chemosphere 9: 551-558. - - - - , 1980. Accumulation and elimination of ~C-3~-HCH (lindane) in Nereis virens (polychaeta) with consideration of metabolism.--Helgolfinder Meeresunters. 33: 313-326. HAMELINK, J.L., R.C. WAYBRANT & R.C. BALL, 1971. A proposal: exchange equilibria control the degree chlorinated hydrocarbons are biologically magnified in bentic environments.--Trans. Am. Fish. Soc. 100: 207-214. HARDINC, G.C.H. & W.P. VASS, 1979. Uptake from seawater and clearance ofp,p'DDT by marine planktonic Crustacea.--J. Fish. Res. Bd Can. 36: 247-254. HOLDEN, A.V. & K. MARSDEN, 1969. Single-stage clean-up of animal tissue extracts for organochlorine residue analysis.--J. Chromat. 44: 481-492. JANSSON, B., S. JENSEN, L. OLSSON,L. RENBERG, G. SUNDSTR()M& R. VAZ, 1975. Identification by GC-MS of phenolic metabolites of PCB and p,p'-DDE isolated from Baltic guillemot and seal.--Ambio 4 (2): 93-97. KOEMAN, J . H . & H. VAN GENDEREN, 1966. Some preliminary notes on residues of chlorinated hydrocarbons insecticides in birds and mammals in the

38

J . C . D U I N K E R ET AL.

Netherlands.--,J. appl. Ecol. 3 (Suppl.): 99-106. - - - - , 1972. Tissue levels in animals and effect caused by chlorinated hydrocarbon insecticides, chlorinated biphenyls and mercury in the marine environment along the Netherlands coast. Mar. Poll. sea life, Fish. News: 1-8. LANOSTON, W.J., 1978. Accumulation of polychlorinated biphenyls in the cockle Cerastoderma edule and the tellinid Macoma balthica.--Mar. Biol. 45: 265-272. McLEESE, D.W., H.C.D. METCALFE & D.S. PEZZACK, 1980. Uptake of PCB's from sediment by Nerds virens and Crangon septumspinoza--Arch, env. contam. Toxicol. 9 (5): 507-518. McLEESE, D.W., S. RAY & C.D. METCALI"E, 1979. Chlorinated hydrocarbon pesticides and polynuclear aromatic hydrocarbons in seditnents and invertebrates from three coastal areas in N e w Brunswick, Canada, a natural bioassay. ICES C . M . 1979/E 28. LEE, R.F., 1981. Mixed function oxidase in marine invertebrates.--Mar. Biol. I,ett. 2 (2): 87-105. PAl., D., J.B. WEBER & M . R . OVERCASH, 1980. Fate of polychlorinated biphenyls (PCB's) in soil-plant systems.--Residue Rev. 74: 45-98. PAVLOU, S.P. ~¢ R.N. DEXTER, 1979. Distribution of polychlorinated biphenyl (PCB) in estuarine ecosystems. Testing the concept of equilibrium partitioning in the marine environment.--Environ. Sci. Technol. 13 (1)" 65-71. PLAGMAN, J., 1939. Ern~ihrungsbiologie der Garnele (Crangon vulgaris Fabr.).-Helgol/inder wiss. Meeresunters. 2 (1): 113-162. REqNDERS, P.J.H., 1980. Organochlorine and heavy metal residues in harbour seals from the Wadden Sea and their possible effects on reproduction.--Neth..]. Sea Res. 14 (1): 30-65. STEEN, W.C., D.F. PARIS & G.L. BAUGHMAN, 1978. Partitioning of selected polychlorinated biphenyls to natural sediments.--Water Res~ 12: 655-657. SUNDSTROM, G., O. HUTZINGER & S. SAFE, 1976. The metabolism of chlorobiphenyls--A review.--Chemosphere 5: 267-298. SWENNEN, C., 1972. Chlorinated hydrocarbons attacked the Eider population in the N e t h e r l a n d s . - - T . N . O . Nieuws 27: 556-560. UREY, J . C . , J . C . KRICHER & J . M . BOYLAN, 1976. Bioconcentration of tbur pure PCB isomeres by Chlorella pyrenoidosa.--Bull, env. contain. Toxicol. (U.S.) 16 (1): 81-85. VERWE¥, J., 1978. De eenheid van milieu en organismen in de Waddenzee: een overzicht. Mededeling van de Werkgroep Waddengebied 4: 1-160. WILDISU, D.J., C.D. METCALVE, H . M . AGAKI & D.W. McLEEsE, 1980. Flux of Aroclor 1254 between estuarine sediment and water.--Bull, env. contain. Toxicol. (U.S.) 24: 20-26. YOUNG, G.R., J.A. SWADER & S.W. BmNGHAM, 1980. Kepone uptake by algae and effects on growth, photosynthesis and respiration.--Va J. Sci. 31 (3): t7-54. Z~MMERMAN,J . T . F . , 1976. Mixing and flushing of tidal ernbayments in the western Wadden Sea. Brill, Leiden (thesis).