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J.Health Sci., 51(2), 178-184, 2005
Determination of Estrogenic Substances in the Water of Muko River Using
in Vitro Assays, and the Degradation of Natural Estrogens by Aquatic Bacteria
Sumiko Matsuoka,* Misa Kikuchi, Sachi Kimura, Yuko Kurokawa, and Shin'ichiro Kawai
School of Human Sciences, Kobe College, Okadayama 4-1, Nishinomiya, Hyogo 662-8505, Japan
In order to determine the level of estrogenic substances in river water and to evaluate the degradation of
estrogen by aquatic bacteria, three in
vitro assays, E-Screen, Ishikawa cell-alkaline phosphatase (Ishikawa
cell-ALP) assay and yeast estrogen screen (YES) assay were used. Water samples were collected throughout one
year at upstream, midstream and downstream locations on the Muko River, Hyogo Prefecture, Japan. Estrogenic
substances in the water were extracted by solid phase extraction using Sep-pak C18 cartridges. The levels of
estrogenic substances in river water changed daily, weekly and monthly. The highest
17beta-estradiol equivalent (E2 equivalent) was obtained as 32.9 ng/l at the midstream in July of 2002. Although some differences in E2
equivalent were observed in some water samples between the E-Screen and Ishikawa cell-ALP assays, the levels
measured by these assays were generally similar and ranged from not detected (ND) to 32.9 ng/l. E2 equivalent
levels assayed by YES were low or below the detection limit. Degradability of estrogen by aquatic bacteria was
investigated in the summer and winter. E2 and estrone (E1) were degraded completely within five days in the
summer, and within seven days in winter. Bacterial degradation of a synthetic estrogen,
17alpha-ethynylestradiol (EE2), was much lower compared to that of E2 or E1.
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