Abstract
Environmental agencies must monitor an ever increasing range of contaminants of emerging concern, including endocrine disrupting compounds (EDCs). An alternative to using ultra-trace chemical analysis of samples for EDCs is to test for biological activity using in vitro screening assays, then use these assay results to direct analytical chemistry approaches. In this study, we used both analytical approaches and in vitro bioassays to characterize the EDCs present in treated wastewater from four wastewater treatment plants (WWTPs) in Ontario, Canada. Estrogen-mediated activity was assessed using a yeast estrogenicity screening (YES) assay. An in vitro competitive binding assay was used to assess capacity to interfere with binding of the thyroid hormone, thyroxine (T4) to the recombinant human thyroid hormone transport protein, transthyretin (i.e. hTTR). An in vitro binding assay with a rat peroxisome proliferator responsive element transfected into a rainbow trout gill cell line was used to evaluate binding and subsequent gene expression via the peroxisome proliferator activated receptor (PPAR). Analyses of a suite of contaminants known to be EDCs in extracts from treated wastewater were conducted using either gas chromatography with mass spectrometry (GC-MS) or liquid chromatography with tandem mass spectrometry (LC-MS/MS). Estrogenic activity was detected in the YES assay only
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Metcalfe, Chris D., E-mail: cmetcalfe@trentu.ca;
[1]
Kleywegt, Sonya;
[2]
Letcher, Robert J.;
[3]
Topp, Edward;
[4]
Wagh, Purva;
Trudeau, Vance L.;
Moon, Thomas W.
[5]
- Environmental and Resource Studies, Trent University, Peterborough, ON, K9J 7B8 (Canada)
- Standards Development Branch, Ontario Ministry of the Environment, 40 St. Clair Ave. West, Toronto, ON, M4V 1M2 (Canada)
- Ecotoxicology and Wildlife Health Division, Science and Technology Branch, Environment Canada, National Wildlife Research Centre, Carleton University, Ottawa, ON, K1A 0H3 (Canada)
- Agriculture and Agri-Food Canada, Southern Crop Protection and Food Research Centre, London, ON, N5V 7T3 (Canada)
- Department of Biology and Centre for Advanced Research in Environmental Genomics, University of Ottawa, Ottawa, ON, K1N 6N5 (Canada)
Citation Formats
Metcalfe, Chris D., E-mail: cmetcalfe@trentu.ca, Kleywegt, Sonya, Letcher, Robert J., Topp, Edward, Wagh, Purva, Trudeau, Vance L., and Moon, Thomas W.
A multi-assay screening approach for assessment of endocrine-active contaminants in wastewater effluent samples.
Netherlands: N. p.,
2013.
Web.
doi:10.1016/J.SCITOTENV.2013.02.074.
Metcalfe, Chris D., E-mail: cmetcalfe@trentu.ca, Kleywegt, Sonya, Letcher, Robert J., Topp, Edward, Wagh, Purva, Trudeau, Vance L., & Moon, Thomas W.
A multi-assay screening approach for assessment of endocrine-active contaminants in wastewater effluent samples.
Netherlands.
https://doi.org/10.1016/J.SCITOTENV.2013.02.074
Metcalfe, Chris D., E-mail: cmetcalfe@trentu.ca, Kleywegt, Sonya, Letcher, Robert J., Topp, Edward, Wagh, Purva, Trudeau, Vance L., and Moon, Thomas W.
2013.
"A multi-assay screening approach for assessment of endocrine-active contaminants in wastewater effluent samples."
Netherlands.
https://doi.org/10.1016/J.SCITOTENV.2013.02.074.
@misc{etde_22277169,
title = {A multi-assay screening approach for assessment of endocrine-active contaminants in wastewater effluent samples}
author = {Metcalfe, Chris D., E-mail: cmetcalfe@trentu.ca, Kleywegt, Sonya, Letcher, Robert J., Topp, Edward, Wagh, Purva, Trudeau, Vance L., and Moon, Thomas W.}
abstractNote = {Environmental agencies must monitor an ever increasing range of contaminants of emerging concern, including endocrine disrupting compounds (EDCs). An alternative to using ultra-trace chemical analysis of samples for EDCs is to test for biological activity using in vitro screening assays, then use these assay results to direct analytical chemistry approaches. In this study, we used both analytical approaches and in vitro bioassays to characterize the EDCs present in treated wastewater from four wastewater treatment plants (WWTPs) in Ontario, Canada. Estrogen-mediated activity was assessed using a yeast estrogenicity screening (YES) assay. An in vitro competitive binding assay was used to assess capacity to interfere with binding of the thyroid hormone, thyroxine (T4) to the recombinant human thyroid hormone transport protein, transthyretin (i.e. hTTR). An in vitro binding assay with a rat peroxisome proliferator responsive element transfected into a rainbow trout gill cell line was used to evaluate binding and subsequent gene expression via the peroxisome proliferator activated receptor (PPAR). Analyses of a suite of contaminants known to be EDCs in extracts from treated wastewater were conducted using either gas chromatography with mass spectrometry (GC-MS) or liquid chromatography with tandem mass spectrometry (LC-MS/MS). Estrogenic activity was detected in the YES assay only in those extracts that contained detectable amounts of estradiol (E2). There was a positive relationship between the degree of response in the T4-hTTR assay and the amounts of polybrominated diphenyl ether (PBDE) congeners 47 and 99, triclosan and the PBDE metabolite, 4-OH-BDE17. Several wastewater extracts gave a positive response in the PPAR assay, but these responses were not correlated with the amounts of any of the EDCs analyzed by LC-MS/MS. Overall, these data indicate that a step-wise approach is feasible using a combination of in vitro testing and instrumental analysis to monitor for EDCs in wastewater and other environmental matrixes. - Highlights: • Tested wastewater for in vitro activity for comparison to analytical measurements of EDCs. • Estrogenic activity was detected only in extracts that contained estradiol or estrone. • Positive relationship observed for binding to thyroid transport protein and 4-OH-PBDE17. • Wastewater showed response in peroxisome proliferator receptor assay, but no relationship observed with levels of contaminants. • Feasible to use in vitro testing as a screening tool before instrumental analysis of contaminants.}
doi = {10.1016/J.SCITOTENV.2013.02.074}
journal = []
volume = {454-455}
journal type = {AC}
place = {Netherlands}
year = {2013}
month = {Jun}
}
title = {A multi-assay screening approach for assessment of endocrine-active contaminants in wastewater effluent samples}
author = {Metcalfe, Chris D., E-mail: cmetcalfe@trentu.ca, Kleywegt, Sonya, Letcher, Robert J., Topp, Edward, Wagh, Purva, Trudeau, Vance L., and Moon, Thomas W.}
abstractNote = {Environmental agencies must monitor an ever increasing range of contaminants of emerging concern, including endocrine disrupting compounds (EDCs). An alternative to using ultra-trace chemical analysis of samples for EDCs is to test for biological activity using in vitro screening assays, then use these assay results to direct analytical chemistry approaches. In this study, we used both analytical approaches and in vitro bioassays to characterize the EDCs present in treated wastewater from four wastewater treatment plants (WWTPs) in Ontario, Canada. Estrogen-mediated activity was assessed using a yeast estrogenicity screening (YES) assay. An in vitro competitive binding assay was used to assess capacity to interfere with binding of the thyroid hormone, thyroxine (T4) to the recombinant human thyroid hormone transport protein, transthyretin (i.e. hTTR). An in vitro binding assay with a rat peroxisome proliferator responsive element transfected into a rainbow trout gill cell line was used to evaluate binding and subsequent gene expression via the peroxisome proliferator activated receptor (PPAR). Analyses of a suite of contaminants known to be EDCs in extracts from treated wastewater were conducted using either gas chromatography with mass spectrometry (GC-MS) or liquid chromatography with tandem mass spectrometry (LC-MS/MS). Estrogenic activity was detected in the YES assay only in those extracts that contained detectable amounts of estradiol (E2). There was a positive relationship between the degree of response in the T4-hTTR assay and the amounts of polybrominated diphenyl ether (PBDE) congeners 47 and 99, triclosan and the PBDE metabolite, 4-OH-BDE17. Several wastewater extracts gave a positive response in the PPAR assay, but these responses were not correlated with the amounts of any of the EDCs analyzed by LC-MS/MS. Overall, these data indicate that a step-wise approach is feasible using a combination of in vitro testing and instrumental analysis to monitor for EDCs in wastewater and other environmental matrixes. - Highlights: • Tested wastewater for in vitro activity for comparison to analytical measurements of EDCs. • Estrogenic activity was detected only in extracts that contained estradiol or estrone. • Positive relationship observed for binding to thyroid transport protein and 4-OH-PBDE17. • Wastewater showed response in peroxisome proliferator receptor assay, but no relationship observed with levels of contaminants. • Feasible to use in vitro testing as a screening tool before instrumental analysis of contaminants.}
doi = {10.1016/J.SCITOTENV.2013.02.074}
journal = []
volume = {454-455}
journal type = {AC}
place = {Netherlands}
year = {2013}
month = {Jun}
}