Occurrence of diclofenac and its metabolites in surface water and effluent samples from Karachi, Pakistan.

Diclofenac and five of its transformation products were identified in Malir River and Lyari River water as well as in effluent samples from Karachi, Pakistan. To the best of our knowledge, this is the first report on the identification of 3'-hydroxydiclofenac and 8-chlorocarbazole-1-yl-ethanoic acid in environmental samples. Their estimated concentrations ranged between 0.08-0.3 microg L(-1) and 0.03-0.4 microg L(-1), respectively. In addition, 4'- and 5-hydroxydiclofenac as well as 1-(2,6-dichlorophenyl)-1,3-dihydro-2H-indole-2-one were detected in the samples at concentrations between 0.4-1.8, 0.01-0.3, and 0.02-0.2 microg L(-1), respectively. The hydroxy derivatives seem to reflect human excretions via household wastewater, while the chlorocarbazole derivative is related to abiotic photolytic transformation of diclofenac.

Expression of heat shock protein 70 in a permanent cell line (EPC) exposed to sediment extracts from the North Sea and the Baltic Sea.

Heat shock proteins 70 (hsp70) are known to be induced by a great variety of chemical stressors. The effects of different environmental contaminants, which were identified in sediments of the North Sea and the Baltic Sea as main contaminants or as contaminants of special toxicological interest, on the extent of expression of hsp70 in a permanent cell line Epithelioma papulosum cyprini (EPC) were investigated. Concentrations of the single contaminants leading to elevated hsp70 levels were compared to the respective concentrations of these contaminants encountered in the sediment extracts thus allowing an assessment of their contribution to the toxicity of these sediments. Analysis of hsp70 was performed using western blotting techniques with subsequent comparative quantification by densitometry. Eleven of thirteen contaminants have shown a dose-dependent increase in hsp70 levels. An expression of hsp70 was observed in most cases between 0.1 and 10 microg/mL test solution. Some of the investigated stressors led to extremely high hsp70 contents even at low concentrations. However, in most cases contaminant concentrations in sediments were too low to cause an effect. For this reason, effects of the tested sediments are attributed to other contaminants or rather to mixtures of stressors.

Brominated indoles and phenols in marine sediment and water extracts from the north and baltic seas-concentrations and effects.

This work presents results from analytical as well as ecotoxicologic investigations of sediment and water samples from the North and Baltic Seas. A bioassay-directed procedure was used to investigate cause-effect relationships between observed effects in acute laboratory bioassays (luminescent bacteria assay with Vibrio fischeri and embryo test with Danio rerio) and analyte concentrations in extracted samples. Brominated phenols and indoles-including 4-bromophenol; 2,4-dibromophenol; 4- and 6-bromoindole; 3,4-, 4,6-, and 3,6-dibromoindole; and tribrominated compounds-were identified in partly remarkable concentrations (up to 40,000 ng g(-1) total organic carbon TOC for 4-bromophenol) in North Sea sediment samples and water samples (913 ng L(-1) 3,6-dibromoindole) from the German Bight. The toxicity of some of the identified brominated substances was low, with median effect concentration levels (EC(50)) ranging from 0.08 to 21.7 mg/L for V. fischeri and 4.3 to 46.3 mg/L for D. rerio. Comparison of the concentrations of analytes with ECs showed a toxicity contribution of brominated phenols and indoles to overall toxicity of the fraction. In the case of one water sample from the German Bight, brominated phenols and indoles accounted for the observed toxicity. Brominated phenols and indoles, which are assumed to be of biogenic origin, have rarely been discussed so far in the context of ecotoxicologic effects in marine ecosystems.

Bioassay-directed chemical analysis of River Elbe surface water including large volume extractions and high performance fractionation.

A bioassay-directed fractionation and identification (toxicity identification evaluation procedure) was performed on extracts of 10 1 River Elbe water samples. The experimental method included a SDB-1 solid phase extraction followed by RP-HPLC fractionation and subfractionation. Chemical analysis by GC-MS as well as acute toxicity testing using a luminescent bacteria assay were conducted in the respective fractions. Many substances were identified, among which were pesticides and pharmaceuticals, but many compounds remained unknown.

Synthetic musks in the environment. Part 1: Species-dependent bioaccumulation of polycyclic and nitro musk fragrances in freshwater fish and mussels.

Bioaccumulation of polycyclic musks (HHCB, AHTN) and nitro musks (musk xylene, musk ketone, and their amino metabolites) in aquatic biota was investigated by analyzing 18 fish samples (rudd, tench, crucian carp, eel) and 1 pooled zebra mussel sample from the pond of a municipal sewage treatment plant. Furthermore, water samples taken at the effluent of the sewage plant as well as water samples and two series of semipermeable membrane devices (SPMDs) from the pond were included. This comprehensive data set allowed the determination of species-dependent bioaccumulation factors on a lipid basis (BAF(L)), e.g., for HHCB the BAF(L) in tench were more than 20 times higher than in eel. The BAF(L) for HHCB and AHTN in biota were lower than the partition coefficients K(SPMD/W) obtained from SPMD samples, which are assumed to represent model bioconcentration values. This stresses that metabolism of these compounds in fish must not be neglected.

Synthetic musks in the environment. Part 2: Enantioselective transformation of the polycyclic musk fragrances HHCB, AHTN, AHDI, and ATII in freshwater fish.

A method for the enantioselective separation of the chiral polycyclic musks HHCB, AHTN, AHDI, and ATII is presented. Eighteen fish samples (rudd, tench, crucian carp, eel) and one pooled zebra mussel sample from the pond of a municipal sewage treatment plant were investigated with regard to their concentrations and the enantiomeric ratios (ERs) of polycyclic musks. In addition, three water samples taken at the effluent of the sewage plant, as well as two water samples and two series of semipermeable membrane devices (SPMDs) consisting of six samples each from the pond were included in the present study. This comprehensive data set allowed a reliable evaluation of species-dependent metabolization processes. The pattern of the polycyclic musks in the chromatograms obtained by enantioselective gas chromatography seems to be typical of each species, like a fingerprint. The highest deviations from the racemic ER were found for trans-HHCB and trans-ATII in crucian carp with values of or below 0.1. Calculations showed that enantioselective transformation seems to be the most important process, resulting in the observed lower concentrations in crucian carp compared to tench. Consequences for a risk assessment are discussed.

Seasonal dependence of the chiral composition of alpha-HCH in coastal deposition at the North Sea.

The environmental pollutant alpha-HCH was analysed in seawater, air and bulk deposition samples from the North Sea. Enantioselective analyses of the sample extracts gave evidence of the change of the direction of net air-water gas transfer of the contaminant in dependence on the season. Due to warmer water surface temperatures during late summer to early autumn the equilibrium of alpha-HCH between air and water is dominated rather by volatilisation than deposition. The volatilisation of non-racemic alpha-HCH, as known to occur from seawater, changes the enantiomeric ratio in air, which is reflected in the observed ratio in rain that passes the air column.

New method for rapid solid-phase extraction of large-volume water samples and its application to non-target screening of North Sea water for organic contaminants by gas chromatography-mass spectrometry.

A method has been developed that allows the solid-phase extraction of microorganic compounds from large volumes of water (10 l) for non-target analysis of filtered seawater. The filtration-extraction system is operated with glass fibre filter candles and the polymeric styrene-divinylbenzene sorbent SDB-1 at flow-rates as high as 500 ml/min. Recovery studies carried out for a couple of model substances covering a wide range of polarity and chemical classes revealed a good performance of the method. Especially for polar compounds (log Kow 3.3-0.7) quantitative recovery was achieved. Limits of detection were between 0.1 and 0.7 ng/l in the full scan mode of the MS. The suitability of the method for the analysis of marine water samples is demonstrated by the non-target screening of water from the German Bight for the presence of organic contaminants. In the course of this screening a large variety of substances was identified including pesticides, industrial chemicals and pharmaceuticals. For some of the identified compounds their occurrence in marine ecosystems has not been reported before, such as dichloropyridines, carbamazepine, propyphenazone and caffeine.

Chromatographic enantiomer separation of chiral xenobiotics and their metabolites--a versatile tool for process studies in marine and terrestrial ecosystems.

A review discussing methodical aspects of enantioselective chromatographic separation of chiral environmental xenobiotics as well as examples for process studies reported in literature. The process studies include microbial transformation of chiral pollutants in aquatic ecosystems, their enzymatic transformation in biota, their photochemical degradation, air/sea exchange processes and atmospheric long range transport, and enantioselective toxic effects.

Enantiomeric composition of the polycyclic musks HHCB and AHTN in different aquatic species

Synthetic polycyclic musk fragrances are mainly represented by the compounds HHCB (Galaxolide(TM)) and AHTN (Tonalide(TM)). Because of their volume of use and their bioaccumulation potential, there is concern with respect to their environmental safety. HHCB and AHTN are chiral compounds, and gas chromatography using modified cyclodextrins as chiral stationary phases coupled to high-resolution mass spectrometry enabled enantioselective analysis even under unfavorable matrix conditions. The gas chromatographic elution order of (4S,7RS)- and (4R,7RS)-HHCB was assigned using synthetic (4S, 7RS)-HHCB. Fish and mussels reared in a pond associated with a municipal waste water treatment plant and semipermeable membrane devices exposed in the pond were analyzed for HHCB and AHTN. The highest lipid concentrations of HHCB and AHTN were observed in mussels (Dreissena polymorpha), tench (Tinca tinca), and crucian carp (Carassius carassius). Pronounced deviations in enantiomeric composition from racemic HHCB were observed in crucian carp and from racemic AHTN in tench. Correlations between lipid levels, enrichment, and enantioselective biotransformation of HHCB or AHTN were not seen. Selective biotransformation depended on both the compound and the species involved. The present study gives the first account of the enantiomeric composition of HHCB and AHTN in aquatic species. The lactone, 1,3,4,6,7,8-hexahydro-4,6,6,7,8, 8-hexamethylcyclopenta[g]-2-benzopyran-1-one, an oxidation product of HHCB, has been identified for the first time in environmental samples. Copyright 1999 Wiley-Liss, Inc.

Musk xylene and musk ketone amino metabolites in the aquatic environment.

The monoamino metabolites of the nitro musk fragrances musk xylene (MX) and musk ketone (MK) were analysed simultaneously with their parent compounds by GC/ECD, GC/PND and GC/EI/MS in the various compartments of the aquatic environment. In this review the data of the metabolites 4-NH2-MX, 2-NH2-MX, and 2-NH2-MK in five river water and seven sewage samples, six sediment samples and in a total of 33 biota samples are summarized and discussed. In the effluents of two municipal sewage plants low nitro musk concentrations and comparatively high levels of the amino metabolites (maximum concentrations: 34 ng 4-NH2-MX/L, 250 ng 2-NH2-MK/L) were analysed indicating that besides adsorption to the sludge the metabolization pathway plays an important role in the sewage plant. In water samples from the river Elbe the transformation products were the dominant compounds as well. In general, in water samples the concentrations of 2-NH2-MK exceeded those of the main MX metabolite 4-NH2-MX significantly. In biota samples 4-NH2-MX seems to be the main metabolite, very often its contents were higher than those of the parent compound. Maximum concentrations of 4-NH2-MX were found in tenches from a sewage pond (3600 microg/kg lipid), a species dependent bioaccumulation was discussed. The bioconcentration of 2-NH2-MK in biota samples is relatively low. There are only few toxicological studies on the mixed amino nitroaromatics, whose data indicate the relevance of the monoamino metabolites in environmental analysis and toxicology and the urgent need of further investigations.

Chlorostyrenes in fish and sediment samples from the river Elbe.

Chlorinated styrenes were identified in fish (bream) and sediment samples from the river Elbe. Four hexachlorostyrenes, four heptachlorostyrenes (concentrations in fish 1-10 ng/g wet weight) and octachlorostyrene (10-45 ng/g wet weight) were analysed. The gas chromatographic retention times and the electron impact (EI) mass spectra of the respective hepta- and hexachloro congeners found in the environment as well as those of some synthesised compounds are presented and discussed. The comparison of the chromatograms from the samples in the upper part of the river with those from the estuary revealed a possible difference in the pattern of chlorostyrenes.

Enantioselective degradation of alpha-hexachlorocyclohexane and cyclodiene insecticides in roe-deer liver samples from different regions of Germany.

Remarkably high concentrations of alpha-hexachlorocyclohexane (alpha-HCH), cis-heptachlorepoxide and oxychlordane were found in roe-deer liver samples both from the northern and southern German states Schleswig-Holstein and Baden-Württemberg, respectively. The data revealed no significant regional differences, but they showed some common characteristics: a preferential degradation of (+)-alpha-HCH, and a preferential enrichment of (+)-oxychlordane and of (+)-cis-heptachlorepoxide as determined by chiral capillary gas chromatography using modified cyclodextrin phases. Calculation of the spearman rank correlation coefficients rS supported the assumption that higher concentrations of alpha-HCH may result in a stronger decomposition of the (+)-enantiomer, while higher levels of cis-heptachlorepoxide and oxychlordane appear to lead to a faster decomposition of the respective (-)-enantiomer or a preferential formation of the respective (+)-enantiomer.

Enantioselective and nonenantioselective degradation of organic pollutants in the marine ecosystem.

Enantiomeric ratios of 11 chiral environmental pollutants determined in different compartments of the marine ecosystem by chiral capillary gas chromatography and chiral high-performance liquid chromatography allow discrimination between the following processes: enantioselective decomposition of both enantiomers with different velocities by marine microorganisms (alpha-HCH, beta-PCCH, gamma-PCCH); enantioselective decomposition of one enantiomer only by marine microorganisms (DCPP); enantioselective decomposition by enzymatic processes in marine biota (alpha-HCH, beta-PCCH, trans-chlordane, cis-chlordane, octachlordane MC4, octachlordane MC5, octachlordane MC7, oxychlordane, heptachlor epoxide); enantioselective active transport through the "blood-brain barrier" (alpha-HCH); nonenantioselective photochemical degradation (alpha-HCH, beta-PCCH).

Chromatographic separation of marine organic pollutants.

A review and discussion of the chromatographic separation of marine organic pollutants is given, including sampling and clean-up procedures, fractionation and enrichment of marine pollutants, capillary gas chromatography (cGC) and high-performance liquid chromatography applying both classical and chiral stationary phases. The potential of multi-dimensional cGC for the analysis of marine organic trace pollutants is discussed for polychlorinated biphenyls (PCBs). The chromatographic separation of coplanar PCBs and of the enantiomers of chiral pollutants provides a further insight into the toxic potential of these marine organic pollutants.