Water quality — Determination of the estrogenic potential of water and waste water — Part 3: In vitro human cell-based reporter gene assay

This document specifies a method for the determination of the estrogenic potential of water and waste water by means of a reporter gene assay utilizing stably transfected human cells. This reporter gene assay is based on the activation of the human estrogen receptor alpha.

This method is applicable to:

— fresh water;

— waste water;

— aqueous extracts and leachates;

— eluates of sediments (fresh water);

— pore water;

— aqueous solutions of single substances or of chemical mixtures;

— drinking water;

— the limit of quantification (LOQ) of this method for the direct analysis of water samples is between 0,3 ng/l and 1 ng/l 17β-estradiol equivalents (EEQ) based on the results of the international interlaboratory trial (see Annex F). The upper working range was evaluated [based on the results of the international interlaboratory trial (see Table F.3)] up to a level of 75 ng EEQ/l. Samples showing estrogenic potencies above this threshold have to be diluted for a valid quantification. Extraction and pre concentration of water samples can prove necessary if their estrogenic potential is below the given LOQ.