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Gas-liquid chromatography-tandem mass spectrometry methodology for the quantitation of estrogenic contaminants in bile of fish exposed to wastewater treatment works effluents and from wild populations

Fenlon, Kate A.; Johnson, Andrew C.; Tyler, Charles R.; Hill, Elizabeth M.. 2010 Gas-liquid chromatography-tandem mass spectrometry methodology for the quantitation of estrogenic contaminants in bile of fish exposed to wastewater treatment works effluents and from wild populations. Journal of Chromatography A, 1217 (1). 112-118. 10.1016/j.chroma.2009.10.063

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Abstract/Summary

Fish can be exposed to a complex mixture of chemical contaminants arising from the exposure to wastewater treatment works (WwTWs) effluents. Some of these contaminants are estrogenic and have been associated with feminisation of male fish and the presence of populations containing intersex individuals. In order to assess the exposure of fish to feminising contaminants in receiving waters and effluents, an analytical method was developed to quantitate the major estrogenic substances present in fish bile, where many xenobiotics are excreted and concentrated. Estrogenic metabolites in bile were deconjugated using enzymatic hydrolysis and partially purified by solid-phase extraction. Steroidal and xeno- estrogens were derivatised to their trimethylsilyl ethers and quantitated by gas-liquid chromatography-mass spectrometry (GC-MSMS) using multiple reaction monitoring. The method was validated using spiked bile samples from immature female rainbow trout ( as well as bile from sexually mature roach (Rutilus rutilus) that had been exposed to either tap water or an undiluted estrogenic effluent for 10 days or captured from a river site downstream of a WwTWs effluent discharge. The mean recovery of target analytes from spiked bile was between 86.4-99.2% and the limit of detection was between 0.1-0.7 ng/ml bile for bisphenol A (BPA), 17β-estradiol E2), estrone (E1) and 17α-ethinylestradiol (EE2), 11, 60 and 327 ng/ml bile for branched nonyl chain isomeric mixtures of 4-nonylphenolethoxylate (NP1EO), 4-nonylphenol (NP) and 4-nonylphenoldiethoxylate (NP2EO) respectively. All target analytes were detected in bile from roach (mixed sex) exposed directly to a WwTWs effluent with mean concentrations between 8-108 µg/mL bile for nonylphenolics, 0.8-1.3 µg/mL for BPA, E2 and E1 and 21 ng/mL for EE2. With the exception of NP2EO all analytes were detected in at least 2 out of the 5 fish sampled from the River Thames at concentrations between 1-984 ng/mL. BPA and NPO1EO were detected in all three reference fish held in tap water indicating contamination from laboratory plastics. The work shows that analysis of 20-100 µL quantities of bile could be a useful approach in detecting exposure to mixtures of estrogenic contaminants taken up by fish from WwTW effluents and has the potential for monitoring the efficacy of remediation strategies that may be adopted for reduction of these endocrine disrupting chemicals in the environment.

Item Type: Publication - Article
Digital Object Identifier (DOI): 10.1016/j.chroma.2009.10.063
Programmes: CEH Topics & Objectives 2009 - 2012 > Water > WA Topic 2 - Ecohydrological Processes > WA - 2.1 - Identify and quantify sources, fluxes and pathways of water, chemicals ...
CEH Sections: Acreman
ISSN: 0021-9673
NORA Subject Terms: Ecology and Environment
Health
Date made live: 20 Apr 2010 13:42
URI: http://nora.nerc.ac.uk/id/eprint/9246

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