HA6 Endocrine-Disrupting Chemicals in Wastewater Treatment Effluents
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() Removal of Endocrine Disrupting Activity from Water Reclamation Processess I. Comparing Bioassay Results with Chemical Analyses.

Hemming, J1, Drewes, J2, Ladenburger, S2, Mieritz, M1, Barman, M1, Schauer, J1, Sonzogni, W1, 1 Wisconsin State Laboratory of Hygiene, Madison, WI, USA2 Colorado School of Mines, Golden, CO, USA

ABSTRACT- In order to understand the removal of endocrine disrupting chemicals throughout wastewater treatment, it is important to determine the potential of compounds in wastewater to affect biological activity as well as determine which specific compounds are responsible for the observed biological effect. For this reason, both bioassays and chemical analyses are important tools when assessing the ability of specific treatment processes to remove endocrine disrupting chemicals. We have collected samples from twelve water reclamation facilities throughout the United States and used solid phase extraction to concentrate the samples. Two chemical methods were used to quantify specific chemicals. Enzyme-linked immunosorbent assays (ELISAs) after HPLC clean-up were used to determine testosterone, 17-estradiol and estriol concentrations. GC-NCI-MS was used to quantify several steroidal hormones (testosterone, 17-estradiol, 17-ethinylestradiol, estrone, estriol) and phenolic compounds (bisphenol A, 4-tert-octylphenol, nonylphenol). To evaluate estrogenic activity, we used the MCF-7 breast cancer cell proliferation assay (E-screen) and an estrogen receptor transfected yeast assay. An androgen receptor (AR) transfected MCF-7 cell proliferation assay (A-screen) and an AR transfected yeast assay were used to assess androgen activity. The results of these assays, expressed in estrogen or androgen equivalents, agreed reasonably well in assessing the general removal efficiencies of the treatment plant operations. E-screen results from influent samples and samples after primary treatment indicate estrogenic activity ranging from 20 ng/L to near 100 ng/L 17-estradiol equivalents (Eeq). According to the chemical analyses, the vast majority of the activity is due to three hormones (17-estradiol, estrone and estriol) rather than phenolic compounds. Samples collected after secondary treatment generally indicate greater than 90% removal. After secondary treatment, the E-screen results ranged from 0.2 - 8 ng/L Eeq. The ELISA and GC-MS results agreed well in terms of the substantial removal after secondary treatment, although the specific concentrations were not always consistent.

Key words: ELISA, endocrine disruptors, bioassay, GC-NCI-MS

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