A new concept for the bioeffects-related analysis of xenoestrogens: Hyphenation of receptor assays with LC-MS

The wide variety of known and unknown endocrine disrupters demands highly efficient methods for detecting the relevant endocrine disrupting pollutants. Bioeffects-related analysis is expected to solve this problem, combining biomolecular recognition processes with chemical analysis, which therefore reduces costs of instrumental chemical analysis to relevant samples. The hyphenation of both steps provides information on potential bioeffects, structure and concentration of the substances of interest. In this work the hyphenation of an enzyme linked receptor assay (ELRA) with liquid chromatography-tandem mass spectrometry (LC-MSMS) for the analysis of estrogens and xenoestrogens is proposed. Both test systems have been validated with standards and spiked real water samples. The ELRA achieved a detection limit of 0.1 mu g/L for 17 beta-estradiol (without preconcentration). With the LC-MSMS method, steroids could be detected down to 1 ng/L (with preconcentration). Two concepts for a tighter hyphenation of biomolecular interaction and chemical analysis are discussed: the on-line coupling of receptor-affinity chromatography (RAC) to LC-MSMS and the direct measurement of receptor-ligand complexes by LC-MSMS.