To assess the estrogenic activity potentially stemmingfrom 17
-estradiol (E2) in drinking water, ESI-LC-MS wasused to identify the products of its aqueous chlorination underthe following conditions: 50
g/L E2, 1.46 mg/L sodiumhypochlorite, pH 7.5, 25
C. Seven products, inc
luding 2,4-dichloro-17
-estradiol, monochloroestrone, 2,4-dichloroestrone,
and the four byproducts such as 4-[2-(2,6-dichloro-3-hydroxyphenyl)ethyl]-7
-methyloctahydroinden-5-one(product C in the text) were identified in chlorinated E2so
lution. The estrogenic activities of the aqueous chlorinatedE2 so
lution at 10, 30, 60, 120,
and 180 min contact timewere assessed by a yeast two-hybrid system based on thelig
and-dependent interaction of two proteins, a humanestrogen receptor (ER)
and a coactivator. All five so
lutionselicited transcriptional activation induction. The maximal
-galactosidase activities induced by the chlorinated so
lutionat 10, 30,
and 60 min were similar
and slightly lowerthan those before chlorination, while the activities of thechlorinated so
lution at 120
and 180 min were about 40% ofthose before chlorination. Finally, 4-chloro-17
-estradiol (4-chloro-E2) (we failed to synthesize the 2-chloroestrone (2-chloro-E1)), 2,4-dichloro-17
-estradiol (2,4-dichloro-E2),
and2,4-dichloroestrone (2,4-dichloro-E1) were synthesized,
and product C was fractionated by HPLC. It was found that4-chloro-E2 elicited strong estrogenic activity, at almostthe same level as that of estrone (EC50 = 10
2 nM), while 2,4-dichloro-E2 elicited weaker
-galactosidase activitycompared with that of 4-chloro-E2. The EC50 was ca. 10
3nM. The maximal
-galactosidase activity for 2,4-dichloro-E1 was lower than that of 2,4-dichloro-E2, while its EC50was similar to that of 2,4-dichloro-E2. In addition, product C,4-[2-(2,6-dichloro-3-hydroxyphenyl)ethyl]-7
-methyloctahydroinden-5-one, induced high
-galactosidaseactivity at the relatively higher concentration of 3.5 × 10
5nM. On the basis of the dose-response curve of asingle byproduct of chlorinated E2, the estrogenic activityat 120
and 180 min appears to be induced mainly by 2,4-dichloro-E2
and 2,4-dichloro-E1.