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The estrogenic effect of chlorinated water increased even without residual chlorine. The components that can be called "estrogenic effect formation potential" and "estrogenic effect intermediates" are defined in chlorinated humic acid solution. This understanding is very similar to that of the process of trihalomethanes (THMs) formation. An appropriate procedure to detect the estrogenic effect formation potential, which represents the maximum value of estrogenic effect, was demonstrated. On the other hand, organic matters of which estrogenic effect increase or decrease by chlorination are present in natural water. The important point is that the estrogenic effect of chlorinated drinking water is the sum of the increased activity and the decreased activity. An appropriate procedure that concentrates the increased estrogenic activity and the decreased activity in chlorinated drinking water simultaneously was established. The estrogenic effect formation potential of chlorinated Lake Biwa water was measured according to the procedure established in this study. Includes 26 references, figures.