The Science o f the Total Environment, 10 (1978) 219--230 © Elsevier Scientific Publishing Company, Amsterdam -- Printed in The Netherlands
C H L O R I N A T E D A R O M A T I C H Y D R O C A R B O N S IN FISH FROM AN A R E A P O L L U T E D BY I N D U S T R I A L E F F L U E N T S
ELIZABETH BAUMANNOFSTAD, GULBRAND LUNDE ANDKARIMARTINSEN
Ce n tral Institute for Industrial Research, Blindern, Oslo 3 (Norway)
BRAGE RYGG Norwegian Institute for Water Research, Blindern, Oslo 3 (Norway)
(Received March 30th, 1978)
ABSTRACT
The content of chlorinated fat-soluble aromatic hydrocarbons was determined in fish from an area polluted by industrial effluents. The fish species investigated were selected among those used for human consumption. For some samples, both the fillet and liver were investigated. For others the whole fish was used. The following compounds were analysed and quantified: Trichlorobenzene, tetrachlorobenzene, pentachlorobenzene, hexachiorobenzene, heptachlorostyrene, octachlorostyrene and polychlorinated biphenyls. The results indicate an appreciable accumulation in fish of the higher chlorinated compounds as pentachlorobenzene, hexachlorobenzene, heptachlorostyrene and octachiorostyrene. Other chlorinated hydrocarbons as decachlorobiphenyl, pentachloronaphthalene, hexachloronaphthalene and hexachlorostyrene were identified, but not quantified. The total content of fat-soluble chlorine was determined in some samples before and after sulphuric acid treatment. The content of chlorine in the identified and quantified compounds accounted for between 40 and 100 per cent of the total amount of chlorine present in the samples as persistent (sulphuric acid resistant) compounds.
INTRODUCTION
D u r i n g t h e r e c e n t years, it has been realised t h a t t h e r e are n u m e r o u s s o u r c e s f o r c h l o r i n a t e d o r g a n i c c o m p o u n d s o t h e r t h a n t h e use o f c h l o r i n a t e d pesticides a n d certain industrial chemicals like PCB. These include the wastes and b y - p r o d u c t s f r o m industrial processes like PVC p r o d u c t i o n , t h e socalled EDC-tar, and waste f r o m h i g h - t e m p e r a t u r e processes w h e r e o r g a n i c material a n d c h l o r i n e are present. Likewise t h e chlorine bleaching in t h e p a p e r indust r y and chloride d i s i n f e c t i o n in w a t e r t r e a t m e n t are sources f o r this t y p e o f p o l l u t a n t and t h e y usually f o r m a very c o m p l e x m i x t u r e o f d i f f e r e n t
220 chlorinated compounds where normally few are known. In order to characterize such a mixture, it is also of interest to know the extent to which these unknown compounds are present in the total amount of chlorinated compounds present in a sample. A technique for determining the total amount of halogenated organics based on neutron-activation may be used for this type of investigation. It has previously been shown that chlorinated compounds formed in the production of magnesium, by reaction between chlorine and carbon, are released into the Frierfjord and taken up by fish [1]. Analyses of sprat from this fjord and neighbouring fjords have shown that it accumulates trichlorobenzene (3CB), tetrachlorobenzene (4CB), pentachlorobenzene (5CB) and hexachlorobenzene (HCB) as well as heptachlorostyrene (HCS) and octachlorostyrene (OCS). These investigations showed that 5CB, HCB, HCS and OCS were present at rather high levels. Previously OCS has been found in birds [2] and fish [3--6] while HCB has also been found in fish [4--6]. In those investigations where the compounds were quantified the level, however, was rather low. The object of this investigation was to expand the sprat studies by including other fish species from the same area, particularly those species used for human consumption. It was of special interest to investigate to what extent the different chlorinated compounds are accumulated in the different species. In this connection, samples of water and sediment were also collected. In 1975 the discharge of chlorinated compounds was reduced during the summer, and it was therefore of interest to see if this change in the release would lead to a lower level of the chlorinated compounds in the fish.
EXPERIMENTAL
Sample material The following species of fish were selected for the analysis: cod (Gadus morrhua), coalfish (Gadus virers), whiting (Gadus merlangus), pollack (Gadus pollachius), plaice (Pleuronectes platessa), and eel (Anguilla vulgaris), as well as some samples of sprat (Clupea sprattus). Samples of liver were taken from coalfish, cod and pollack. Water and sediment samples were collected from places near to where the chlorinated pollutants are discharged into the fjord.
EXTRACTION A N D CLEAN-UP P R O C E D U R E S For the analysis, either whole fish or fish fillets(and liver)were taken. For each sample, between 15 and 250 g were used depending on the fat content. The samples were homogenized and extracted by shaking for three hours at room temperature with cyclohexane/isopropanol (1:1, v/v). The extraction was repeated once. For both extractions, between 60 and 200 ml of cyclo-
221 hexane were used. The extracts were combined and the cyclohexane phase was separated by addition of water. The cyclohexane extract was further washed twice with distilled water to remove inorganic halogen and dried over sodium sulphate. One aliquot of the cyclohexane extract (1--2 ml) was then treated with an equal volume of concentrated sulphuric acid in order to remove the non-persistent organic compounds. A further clean-up of the sulphuric acid resistant extract was performed by column chromatography on an alumina column using cyclohexane as eluting agent. The alumina was previously washed with cyclohexane and methanol, dried at 100 °C and heated at 550 °C. After this activation, the alumina was deactivated by adding 5% by weight of distilled water and allowed to equilibrate for at least 1 day before use. The alumina was dry-packed in a Pasteur-pipette (5 mm i.d.) using a glass wool plug at the b o t t o m . The height of the column packing was a b o u t 5 cm. The sulphuricacid-treated cyclohexane aliquot was transferred to the column and eluted with 3 times its volume of cyclohexane. The eluate was finally concentrated to the initial aliquot volume by evaporating at 50 °C under a gentle stream of nitrogen. The more polar c o m p o u n d s present in the extracts will n o t be eluted under these conditions. Recovery of the analysed c o m p o u n d s in the column clean-up step has been checked by running some samples on a gas chromatagraph both before and after this treatment. No significant losses of any of the c o m p o u n d s could be observed. The fat content of the samples was determined b y weighing after evaporating the solvent. The sediment samples were extracted in the same way as the fish samples. Four-litre samples of water were extracted for two hours under continuous mixing with a magnetic stirrer with 200 ml cyclohexane. After separation of the cyclohexane phase the same clean-up procedure was used as for the fish samples. A sample of cod liver oil with a high level of chlorinated c o m p o u n d s was also treated separately according to Jensen [7] in order to obtain high sensitivity on the gas chromatography/mass spectrometry (GC/MS) analysis. This method, combined with a two-stage column clean-up and fractionation, give the necessary enrichment of the chlorinated hydrocarbons for identification with GC/MS.
Identification and quantification Determination of the chlorinated c o m p o u n d s in the extracts after sulphuric acid t r e a t m e n t was performed by using a Perkin--Elmer 3920 gas chromatograph with an electron capture detector (ECD). The GC conditions were: Column: 2 m 1/8" i.d. stainless steel; stationary phase: 3% SE-30 on Supelcoport; column temperature: Programmed from 130 °C to 230 °C with a rate of 8 °C/min; carrier gas: N 2 , 2 0 ml/min; ECD " m a k e - u p " gas: Ar with 5% CH4, 20 ml/min. In addition to SE-30 a mixture of SP-2250 {1.5%) and SP-2401 (1.95%) was used as stationary phase to facilitate the identification. The same SE-30 column and identical temperature programming was used for the GC/MS
222 analysis. For this analysis, He was used as carrier gas at a flow-rate of 20 ml/min. The chlorinated benzenes and octachlorostyrene were quantified by comparing with the corresponding standards. Standard for heptachlorostyrene was not available. Because hexachlorobenzene and octachlorostyrene have fairly equal response on ECD, the content of heptachlorostyrene was determined by comparing this compound with a hexachlorobenzene standard. Two isomers of tetrachlorobenzene and three isomers of both trichlorobenzene and heptachlorostyrene were found in the samples. The values given for these compounds are a sum of the isomers found in each sample. PCB is a complex mixture of compounds, and by GC separation a pattern of peaks will be found. The pattern will be different for PCB mixtures with different amounts of chlorine incorporated. A Clophen A 60 {Bayer, 60% C1) mixture was used for determination of PCB, because this standard gives the best resemblance with the PCB pattern of the samples. Standard curves were used for quantification of the different compounds.
Activation analysis Extract samples (2 ml), both untreated and sulphuric-acid treated, were analyzed for total content of organic-bound chlorine and bromine by activation analysis. These analyses were carried out at The Norwegian Institute for Atomic Energy. The method has been described previously [8].
RESULTS A N D C O M M E N T S
The results for fish from the Frierfjord are presented in Table 1, and for fish from the Eidangerfjord, a neighbouring fjord, in Table 2. The localization of the two fjords is shown on the map, Fig. 1. In Table 3 the total amount of chlorine after the sulphuric acid treatment is presented for some selected samples, and compared to the amount of chlorine in the identified and quantified compounds.
/
oC~, ~ /~
- •Frierfjord
~']~'/ r~Eidanger~.,/fjord
Fig. 1. Map showing the localization of the Frierfjord and the Eidangerfjord.
223 WATER 0
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}
=II
x
x
OI
•
I1ili !I
',I oll i !I
x
x
x
Fig. 2. ECD gas chromatogram of water sample from the Frierfjord. Analytical conditions are given in the text.
Figures 2--4 show typical gas chromatograms of the non-polar extracts isolated from water, sediment and fish. A chromatogram for one of the fractions of cod liver oil especially prepared for GC/MS analysis, is shown in Fig. 5. All the identified compounds are listed on the chromatogram. Many of the compounds reported in this paper have previously been found in biological samples. The levels found have, however, mostly been very low and it has not been possible to fred the sources of the pollutants. The relatively high levels of these compounds present in the samples from the Frierfjord have made it possible to study both the transport, the bioaccumulation and the internal distribution of a number of chlorinated compounds that are formed by a high-temperature reaction between chlorine and tar-containing material. The same compounds will probably be formed by most processes of this type, even if the reaction conditions like temperature and oxygen supply vary. This means that the presence of compounds such as octachlorostyrene or heptachlorostyrene in fish samples may indicate that chlorinated benzenes and decachlorobiphenyl are also present. Because HCB also originates from other sources, similar conclusions cannot be drawn when HCB is observed in environmental samples. The high values found for many of the chlorinated compounds in the fish samples from the Frierfjord (Table 1) show that these compounds are taken up and accumulated very efficiently' in fish. Furthermore, the concentration
224
SEDIMENT
o
o
o
PCB
m
i
J x
Fig. 3. ECD gas chromatogram of sediment sample from the Frierfjord. Analytical conditions are given in the text.
levels are highest in samples collected near the effluent outlet and decrease with increasing distance from the place of release. This is clearly demonstrated by comparing the levels in Tables 1 and 2 for samples from the Frierfjord and the Eidangerfjord. These results are in accordance with the results previously reported from the same area [1]. By comparing the levels in different fish species in this investigation, it is evident that some species, especially among the gadoid fishes {cod, whiting, and pollack) are different from the others. These species have especially high contents of hexachlorobenzene, heptachlorostyrene, and octachlorostyrene. The levels are considerably higher than the ones previously found in sprat (1). Also the relative levels of the most important chlorinated c o m p o u n d s (HCB, HCS, OCS) vary for the different fish species. While OCS levels are higher than HCB in the cod fishes (especially in liver), the levels are fairly similar in other types of fish as for instance sprat. For eel and plaice, the HCB levels are even higher than the OCS levels in a few samples. Different life patterns and feeding habits of the different fishes will probably determine the a m o u n t of pollutants taken up, while the metabolism and fat content in the tissues will influence the internal distribution of the compounds. It does n o t appear that the close contact between the sediment and fish will influence the levels as the content of pollutants for b o t t o m fish, for example eel or flounder, is lower than the levels found in cod fishes.
225 COD-LIVER
¢ o
o
~= o~
PCB
D
// Y~
O0
x
Fig, 4. ECD gas chromatogram of cod liver sample from the Frierfjord. Analytical conditions are given in the t e x t .
The a m o u n t of chlorine from the identified c o m p o u n d s in the samples makes up from a b o u t 40 to 100 per cent of the total a m o u n t of chlorine b o u n d in the persistent compounds (Table 3). However, also the total a m o u n t of persistent chlorinated c o m p o u n d s decreases in the same manner as for the identified c o m p o u n d s with increasing distance from the effluent outlet. The content of PCB in the samples varies mainly between 1--70 p p m . The highest values have been f o u n d in cod. The varying levels of PCB within the same species probably mean that PCB is released from local sources, or from the river Skienselva, as well as from the atmosphere. The large differences between the species indicate that there exist species-specific differences for PCB uptake and metabolism in the same way as for the other chlorinated hydrocarbons. When comparing the chromatograms of the extracts from sediments and water (Figs. 2 and 3) with extracts from fish samples (Fig. 4), there seems to be a relative increase in the c o n t e n t of the higher chlorinated c o m p o u n d s in the fish. The most probable explanation is that the lower chlorinated c o m p o u n d s are metabolized at a faster rate. Analysis of off extracted from liver and fillet samples from the same fish indicates that the chlorinated c o m p o u n d s accumulate to a larger extent in the liver than in the fillet. This may be due to a slower exchange of pollutants from the liver than from the muscular tissue.
226
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TABLE3 T O T A L A M O U N T O F PERSISTENT F A T S O L U B L E O R G A N I C C H L O R I N E A N D T H E F R A C T I O N (in %) O F THIS C H L O R I N E ORIGINATING F R O M T H E K N O W N H Y D R O C A R B O N S IN FISH F R O M T H E F R I E R F J O R D A N D T H E E I D A N G E R F J O R D
Sample
Locality
Caught date
Total persistent Cl (ppm)
Cod fillet Cod liver Whiting Pollack fillet Pollack liver Plaice Sprat Sprat Pollack fillet Pollack liver Sprat Cod fillet Cod liver
The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Eidangerfjord The Eidangerfjord The Eidangerfjord The Eidangerfjord The Eidangerfjord
Dec. 1975 July 1975 May 1975 May 1975 May 1975 Sept. 1975 May 1975 Sept. 1975 July 1975 July 1975 July 1975 May 1976 May 1976
225 428 244 39 35 57 107 52 13 17 30 13 36
CI from known compounds in % of total persistent C1 66 88 69 98
73 77 39 49 104 94 56 55 41
230
The release of chlorinated compounds to the Frierfjord was drastically reduced during the summer of 1975 because of a change of the industrial process where these compounds are formed. Relatively large amounts of chlorinated compounds are stored in the sediments in the Frie.fjord, and there is some uncertainty as to what e x t e n t they are released to the water. Analysis through the coming years will probably show if the level of chlorinated compounds in the fish is influenced by the new situation of pollution. REFERENCES 1 G. Lunde and E. Baumann Ofstad, Z. Anal. Chem., 282 (1976) 395. 2 M.C. ten Noever de Brauw and J. H. Koeman, Sci. Total Environ., 1 (1972/73) 427. 3 D.W. Kuehl, H. L. Kopperman, G. D. Veith and G. E. Glass, Bull. Environ. Contain. Toxicol., 1 (1972) 338. 4 A.V. Holden, Pestic.Monit. J., 4 (1970) 117. 5 V. Zitko, Bull. Environ. Contain. Toxicol., 6 (1971) 464. 6 J.L. Johnson, D.L. Stalling and J.W. Hogan, Bull. Environ. Contain. Toxicol., 2 (1974) 393. 7 S. Jensen and B. Jansson, Ambio, 5 (1976) 257. 8 G. Lunde and E. Steinnes, Environ. Sci. Technol., 9 (1975) 155.
C H L O R I N A T E D A R O M A T I C H Y D R O C A R B O N S IN FISH FROM AN A R E A P O L L U T E D BY I N D U S T R I A L E F F L U E N T S
ELIZABETH BAUMANNOFSTAD, GULBRAND LUNDE ANDKARIMARTINSEN
Ce n tral Institute for Industrial Research, Blindern, Oslo 3 (Norway)
BRAGE RYGG Norwegian Institute for Water Research, Blindern, Oslo 3 (Norway)
(Received March 30th, 1978)
ABSTRACT
The content of chlorinated fat-soluble aromatic hydrocarbons was determined in fish from an area polluted by industrial effluents. The fish species investigated were selected among those used for human consumption. For some samples, both the fillet and liver were investigated. For others the whole fish was used. The following compounds were analysed and quantified: Trichlorobenzene, tetrachlorobenzene, pentachlorobenzene, hexachiorobenzene, heptachlorostyrene, octachlorostyrene and polychlorinated biphenyls. The results indicate an appreciable accumulation in fish of the higher chlorinated compounds as pentachlorobenzene, hexachlorobenzene, heptachlorostyrene and octachiorostyrene. Other chlorinated hydrocarbons as decachlorobiphenyl, pentachloronaphthalene, hexachloronaphthalene and hexachlorostyrene were identified, but not quantified. The total content of fat-soluble chlorine was determined in some samples before and after sulphuric acid treatment. The content of chlorine in the identified and quantified compounds accounted for between 40 and 100 per cent of the total amount of chlorine present in the samples as persistent (sulphuric acid resistant) compounds.
INTRODUCTION
D u r i n g t h e r e c e n t years, it has been realised t h a t t h e r e are n u m e r o u s s o u r c e s f o r c h l o r i n a t e d o r g a n i c c o m p o u n d s o t h e r t h a n t h e use o f c h l o r i n a t e d pesticides a n d certain industrial chemicals like PCB. These include the wastes and b y - p r o d u c t s f r o m industrial processes like PVC p r o d u c t i o n , t h e socalled EDC-tar, and waste f r o m h i g h - t e m p e r a t u r e processes w h e r e o r g a n i c material a n d c h l o r i n e are present. Likewise t h e chlorine bleaching in t h e p a p e r indust r y and chloride d i s i n f e c t i o n in w a t e r t r e a t m e n t are sources f o r this t y p e o f p o l l u t a n t and t h e y usually f o r m a very c o m p l e x m i x t u r e o f d i f f e r e n t
220 chlorinated compounds where normally few are known. In order to characterize such a mixture, it is also of interest to know the extent to which these unknown compounds are present in the total amount of chlorinated compounds present in a sample. A technique for determining the total amount of halogenated organics based on neutron-activation may be used for this type of investigation. It has previously been shown that chlorinated compounds formed in the production of magnesium, by reaction between chlorine and carbon, are released into the Frierfjord and taken up by fish [1]. Analyses of sprat from this fjord and neighbouring fjords have shown that it accumulates trichlorobenzene (3CB), tetrachlorobenzene (4CB), pentachlorobenzene (5CB) and hexachlorobenzene (HCB) as well as heptachlorostyrene (HCS) and octachlorostyrene (OCS). These investigations showed that 5CB, HCB, HCS and OCS were present at rather high levels. Previously OCS has been found in birds [2] and fish [3--6] while HCB has also been found in fish [4--6]. In those investigations where the compounds were quantified the level, however, was rather low. The object of this investigation was to expand the sprat studies by including other fish species from the same area, particularly those species used for human consumption. It was of special interest to investigate to what extent the different chlorinated compounds are accumulated in the different species. In this connection, samples of water and sediment were also collected. In 1975 the discharge of chlorinated compounds was reduced during the summer, and it was therefore of interest to see if this change in the release would lead to a lower level of the chlorinated compounds in the fish.
EXPERIMENTAL
Sample material The following species of fish were selected for the analysis: cod (Gadus morrhua), coalfish (Gadus virers), whiting (Gadus merlangus), pollack (Gadus pollachius), plaice (Pleuronectes platessa), and eel (Anguilla vulgaris), as well as some samples of sprat (Clupea sprattus). Samples of liver were taken from coalfish, cod and pollack. Water and sediment samples were collected from places near to where the chlorinated pollutants are discharged into the fjord.
EXTRACTION A N D CLEAN-UP P R O C E D U R E S For the analysis, either whole fish or fish fillets(and liver)were taken. For each sample, between 15 and 250 g were used depending on the fat content. The samples were homogenized and extracted by shaking for three hours at room temperature with cyclohexane/isopropanol (1:1, v/v). The extraction was repeated once. For both extractions, between 60 and 200 ml of cyclo-
221 hexane were used. The extracts were combined and the cyclohexane phase was separated by addition of water. The cyclohexane extract was further washed twice with distilled water to remove inorganic halogen and dried over sodium sulphate. One aliquot of the cyclohexane extract (1--2 ml) was then treated with an equal volume of concentrated sulphuric acid in order to remove the non-persistent organic compounds. A further clean-up of the sulphuric acid resistant extract was performed by column chromatography on an alumina column using cyclohexane as eluting agent. The alumina was previously washed with cyclohexane and methanol, dried at 100 °C and heated at 550 °C. After this activation, the alumina was deactivated by adding 5% by weight of distilled water and allowed to equilibrate for at least 1 day before use. The alumina was dry-packed in a Pasteur-pipette (5 mm i.d.) using a glass wool plug at the b o t t o m . The height of the column packing was a b o u t 5 cm. The sulphuricacid-treated cyclohexane aliquot was transferred to the column and eluted with 3 times its volume of cyclohexane. The eluate was finally concentrated to the initial aliquot volume by evaporating at 50 °C under a gentle stream of nitrogen. The more polar c o m p o u n d s present in the extracts will n o t be eluted under these conditions. Recovery of the analysed c o m p o u n d s in the column clean-up step has been checked by running some samples on a gas chromatagraph both before and after this treatment. No significant losses of any of the c o m p o u n d s could be observed. The fat content of the samples was determined b y weighing after evaporating the solvent. The sediment samples were extracted in the same way as the fish samples. Four-litre samples of water were extracted for two hours under continuous mixing with a magnetic stirrer with 200 ml cyclohexane. After separation of the cyclohexane phase the same clean-up procedure was used as for the fish samples. A sample of cod liver oil with a high level of chlorinated c o m p o u n d s was also treated separately according to Jensen [7] in order to obtain high sensitivity on the gas chromatography/mass spectrometry (GC/MS) analysis. This method, combined with a two-stage column clean-up and fractionation, give the necessary enrichment of the chlorinated hydrocarbons for identification with GC/MS.
Identification and quantification Determination of the chlorinated c o m p o u n d s in the extracts after sulphuric acid t r e a t m e n t was performed by using a Perkin--Elmer 3920 gas chromatograph with an electron capture detector (ECD). The GC conditions were: Column: 2 m 1/8" i.d. stainless steel; stationary phase: 3% SE-30 on Supelcoport; column temperature: Programmed from 130 °C to 230 °C with a rate of 8 °C/min; carrier gas: N 2 , 2 0 ml/min; ECD " m a k e - u p " gas: Ar with 5% CH4, 20 ml/min. In addition to SE-30 a mixture of SP-2250 {1.5%) and SP-2401 (1.95%) was used as stationary phase to facilitate the identification. The same SE-30 column and identical temperature programming was used for the GC/MS
222 analysis. For this analysis, He was used as carrier gas at a flow-rate of 20 ml/min. The chlorinated benzenes and octachlorostyrene were quantified by comparing with the corresponding standards. Standard for heptachlorostyrene was not available. Because hexachlorobenzene and octachlorostyrene have fairly equal response on ECD, the content of heptachlorostyrene was determined by comparing this compound with a hexachlorobenzene standard. Two isomers of tetrachlorobenzene and three isomers of both trichlorobenzene and heptachlorostyrene were found in the samples. The values given for these compounds are a sum of the isomers found in each sample. PCB is a complex mixture of compounds, and by GC separation a pattern of peaks will be found. The pattern will be different for PCB mixtures with different amounts of chlorine incorporated. A Clophen A 60 {Bayer, 60% C1) mixture was used for determination of PCB, because this standard gives the best resemblance with the PCB pattern of the samples. Standard curves were used for quantification of the different compounds.
Activation analysis Extract samples (2 ml), both untreated and sulphuric-acid treated, were analyzed for total content of organic-bound chlorine and bromine by activation analysis. These analyses were carried out at The Norwegian Institute for Atomic Energy. The method has been described previously [8].
RESULTS A N D C O M M E N T S
The results for fish from the Frierfjord are presented in Table 1, and for fish from the Eidangerfjord, a neighbouring fjord, in Table 2. The localization of the two fjords is shown on the map, Fig. 1. In Table 3 the total amount of chlorine after the sulphuric acid treatment is presented for some selected samples, and compared to the amount of chlorine in the identified and quantified compounds.
/
oC~, ~ /~
- •Frierfjord
~']~'/ r~Eidanger~.,/fjord
Fig. 1. Map showing the localization of the Frierfjord and the Eidangerfjord.
223 WATER 0
C=
NI
511
}
=II
x
x
OI
•
I1ili !I
',I oll i !I
x
x
x
Fig. 2. ECD gas chromatogram of water sample from the Frierfjord. Analytical conditions are given in the text.
Figures 2--4 show typical gas chromatograms of the non-polar extracts isolated from water, sediment and fish. A chromatogram for one of the fractions of cod liver oil especially prepared for GC/MS analysis, is shown in Fig. 5. All the identified compounds are listed on the chromatogram. Many of the compounds reported in this paper have previously been found in biological samples. The levels found have, however, mostly been very low and it has not been possible to fred the sources of the pollutants. The relatively high levels of these compounds present in the samples from the Frierfjord have made it possible to study both the transport, the bioaccumulation and the internal distribution of a number of chlorinated compounds that are formed by a high-temperature reaction between chlorine and tar-containing material. The same compounds will probably be formed by most processes of this type, even if the reaction conditions like temperature and oxygen supply vary. This means that the presence of compounds such as octachlorostyrene or heptachlorostyrene in fish samples may indicate that chlorinated benzenes and decachlorobiphenyl are also present. Because HCB also originates from other sources, similar conclusions cannot be drawn when HCB is observed in environmental samples. The high values found for many of the chlorinated compounds in the fish samples from the Frierfjord (Table 1) show that these compounds are taken up and accumulated very efficiently' in fish. Furthermore, the concentration
224
SEDIMENT
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i
J x
Fig. 3. ECD gas chromatogram of sediment sample from the Frierfjord. Analytical conditions are given in the text.
levels are highest in samples collected near the effluent outlet and decrease with increasing distance from the place of release. This is clearly demonstrated by comparing the levels in Tables 1 and 2 for samples from the Frierfjord and the Eidangerfjord. These results are in accordance with the results previously reported from the same area [1]. By comparing the levels in different fish species in this investigation, it is evident that some species, especially among the gadoid fishes {cod, whiting, and pollack) are different from the others. These species have especially high contents of hexachlorobenzene, heptachlorostyrene, and octachlorostyrene. The levels are considerably higher than the ones previously found in sprat (1). Also the relative levels of the most important chlorinated c o m p o u n d s (HCB, HCS, OCS) vary for the different fish species. While OCS levels are higher than HCB in the cod fishes (especially in liver), the levels are fairly similar in other types of fish as for instance sprat. For eel and plaice, the HCB levels are even higher than the OCS levels in a few samples. Different life patterns and feeding habits of the different fishes will probably determine the a m o u n t of pollutants taken up, while the metabolism and fat content in the tissues will influence the internal distribution of the compounds. It does n o t appear that the close contact between the sediment and fish will influence the levels as the content of pollutants for b o t t o m fish, for example eel or flounder, is lower than the levels found in cod fishes.
225 COD-LIVER
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The a m o u n t of chlorine from the identified c o m p o u n d s in the samples makes up from a b o u t 40 to 100 per cent of the total a m o u n t of chlorine b o u n d in the persistent compounds (Table 3). However, also the total a m o u n t of persistent chlorinated c o m p o u n d s decreases in the same manner as for the identified c o m p o u n d s with increasing distance from the effluent outlet. The content of PCB in the samples varies mainly between 1--70 p p m . The highest values have been f o u n d in cod. The varying levels of PCB within the same species probably mean that PCB is released from local sources, or from the river Skienselva, as well as from the atmosphere. The large differences between the species indicate that there exist species-specific differences for PCB uptake and metabolism in the same way as for the other chlorinated hydrocarbons. When comparing the chromatograms of the extracts from sediments and water (Figs. 2 and 3) with extracts from fish samples (Fig. 4), there seems to be a relative increase in the c o n t e n t of the higher chlorinated c o m p o u n d s in the fish. The most probable explanation is that the lower chlorinated c o m p o u n d s are metabolized at a faster rate. Analysis of off extracted from liver and fillet samples from the same fish indicates that the chlorinated c o m p o u n d s accumulate to a larger extent in the liver than in the fillet. This may be due to a slower exchange of pollutants from the liver than from the muscular tissue.
226
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TABLE3 T O T A L A M O U N T O F PERSISTENT F A T S O L U B L E O R G A N I C C H L O R I N E A N D T H E F R A C T I O N (in %) O F THIS C H L O R I N E ORIGINATING F R O M T H E K N O W N H Y D R O C A R B O N S IN FISH F R O M T H E F R I E R F J O R D A N D T H E E I D A N G E R F J O R D
Sample
Locality
Caught date
Total persistent Cl (ppm)
Cod fillet Cod liver Whiting Pollack fillet Pollack liver Plaice Sprat Sprat Pollack fillet Pollack liver Sprat Cod fillet Cod liver
The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Frierfjord The Eidangerfjord The Eidangerfjord The Eidangerfjord The Eidangerfjord The Eidangerfjord
Dec. 1975 July 1975 May 1975 May 1975 May 1975 Sept. 1975 May 1975 Sept. 1975 July 1975 July 1975 July 1975 May 1976 May 1976
225 428 244 39 35 57 107 52 13 17 30 13 36
CI from known compounds in % of total persistent C1 66 88 69 98
73 77 39 49 104 94 56 55 41
230
The release of chlorinated compounds to the Frierfjord was drastically reduced during the summer of 1975 because of a change of the industrial process where these compounds are formed. Relatively large amounts of chlorinated compounds are stored in the sediments in the Frie.fjord, and there is some uncertainty as to what e x t e n t they are released to the water. Analysis through the coming years will probably show if the level of chlorinated compounds in the fish is influenced by the new situation of pollution. REFERENCES 1 G. Lunde and E. Baumann Ofstad, Z. Anal. Chem., 282 (1976) 395. 2 M.C. ten Noever de Brauw and J. H. Koeman, Sci. Total Environ., 1 (1972/73) 427. 3 D.W. Kuehl, H. L. Kopperman, G. D. Veith and G. E. Glass, Bull. Environ. Contain. Toxicol., 1 (1972) 338. 4 A.V. Holden, Pestic.Monit. J., 4 (1970) 117. 5 V. Zitko, Bull. Environ. Contain. Toxicol., 6 (1971) 464. 6 J.L. Johnson, D.L. Stalling and J.W. Hogan, Bull. Environ. Contain. Toxicol., 2 (1974) 393. 7 S. Jensen and B. Jansson, Ambio, 5 (1976) 257. 8 G. Lunde and E. Steinnes, Environ. Sci. Technol., 9 (1975) 155.
