Products of aqueous chlorination of 4-nonylphenol and their estrogenic activity.

Seven reaction products (2-chloro-4-nonylphenol [NP], 2,6-dichloro-4-NP, trichlorophenol, 4-propyl-2'-hydroxyphenol, 4-isobutyl-2'-hydroxyphenol, 4-isoamyl-2'-hydroxyphenol, and 4-isopentyl-2'-hydroxyphenol) were identified by gas chromatography/mass spectrometry (GC/MS) in order to assess the estrogenic activity originated from 4-nonylphenol (4-NP) in drinking water. The estrogenic activities of the aqueous chlorinated 4-NP solution at 10, 60, and 120 min chlorination time were assessed by a yeast two-hybrid system based on the ligand-dependent interaction of two proteins, a human estrogen receptor (ER), and a coactivator. It was found that all three solutions inhibited transcriptional activation induction by 4-NP. Further experiments showed that these solutions also inhibited beta-galactosidase induction by 17beta-estradiol. For the solution at 10 min, the inhibition was found to be due to its toxicity, with an inhibition concentration (IC50) of about 10-fold of concentration of chlorinated 4-NP solution, suggesting the existence of some products with higher yeast toxicity than that of the parent 4-NP. Similar inhibition trends were also found in the dose response of the two solutions at 60 and 120 min, with an IC50 of 10-fold concentration. In these cases, the effects were considered to result from their antagonist action because the two solutions show lower yeast toxicity of which IC50 is 45-fold concentration. This suggests that some products in the chlorinated 4-NP solution elicit the antiestrogenic activities.