Determinations of estrogens in water using novel reverse osmosis enrichment method with subsequent
in vitro analysis


                             C. Wangmo, E. Rozmánková, and M. Bittner*

               Masaryk University, RECETOX, Kamenice 5, CZ62500 Brno, Czech Republic


            *Corresponding author: E-mail: bittner@recetox.muni.cz, Tel +420 549493807


Abstract

Various compounds in the aquatic environment with potential estrogenicity may be present at very
low (<ng/L) individual concentrations. Accurate and reliable information on the overall estrogenic
effect in the complex matrix can be obtained if the whole water sample is analyzed. Present study
describes applicability of a novel benchtop device based on reverse osmosis (RO) process that was
used to increase the concentration of the estrogenic compounds (17β-estradiol, 17α-ethinyl
estradiol, estrone and estriol) in spiked tap water. Estrogenicity in RO-enriched whole water
samples was measured using optimized in vitro bioassay based on transgenic hERα-HeLa-9903 cell
line. The effectiveness of RO enrichment method was assessed by calculating recovery at ten-fold
enrichment level. Observed recovery of 47 to 79 percent suggests that the novel RO device is
promising method providing enriched whole water samples suitable for subsequent in vitro analysis.

Keywords: reverse osmosis; endocrine disrupting compounds; sample concentration; in vitro bioassay