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2L - Immunotoxicity - genotoxicity - ED (WEP/99) Rapid determination of endocrine disrupting compounds (EDCs) in in vivo and in vitro biological assays. Pojana, Giulio1, Bonfà, Angela1, Collarin, Anna 1, Marcomini, Antonio1, 1 Department of Environmental Sciences, University of Venice, Venice, Italy ABSTRACT- Endocrine disrupting compounds (EDCs) are receiving an increasing attention due to their wide occurrence in the aquatic environment and their potential hazard to aquatic organisms. While field investigations can highlight effects on aquatic life, laboratory experiments, such as in vivo and in vitro biological tests, can quantify estrogenic potency of tested chemicals. The nominal concentrations of the tested chemicals in biological experiments often differ considerably from actually measured concentrations, thus giving rise to a significant source of error in the determination of the correct dose-response curve. The large variety of chemical structures of EDCs, the complexity of matrices and low volumes employed for in vitro experiments, as well as the high number of samples needed for statistically reliable results, require an adequate analytical approch. New methods have been developed for a rapid (up to 10 minutes analysis time), high-through-put (isocratic conditions) determination of actual concentrations of natural (genistein) and synthetic (nonylphenol, nonylphenol mono- and diethoxylates, nonylphenol monoethoxylate carboxylate, bisphenol-A, benzophenone) EDCs in in vivo and in vitro biological assays by HPLC coupled with Ion Trap Mass Spectrometry. The high sensitivity and selectivity of the MS detector allowed a reliable quantification of selected analytes simply by direct injection, thus avoiding any sample pretreatment, in the chromatographic system. The method quantification limits (MQLs) were in the 0.05-10 Key words: biological assay, endocrine disrupting compounds, HPLC-MS |
g/L, matching the lowest concentration levels used for response-dose curve experiments. The developed methods can moreover be applied to EDC mixtures without decrease of analytical performance.