In vivo tests are not commonly used in effect directed analysis (EDA) approaches. In the present study, a novel methodology was developed whereby Potamopyrgus antipodarum, which is known to be sensitive to endocrine disrupting compounds, was used as test organism. Field sediments from a polluted site in the north of Belgium were extracted and fractionated using three coupled and automatically switched normal-phase HPLC columns. Part of the fractions were spiked to artificial sediments and tested in a sediment contact test with P. antipodarum. The other part was used for an in vitro effect confirmation with the ER-LUC and anti-AR CALUX assays. Two of the six tested fractions stimulated the reproduction of the snails, while two others inhibited the reproduction. The fractions that caused an increase in reproduction also showed an increased estrogenic potency in the ER-LUC assay. Chemical analysis revealed that one of the most prominent compounds in those fractions was bisphenol-A, which has already been reported to have a stimulating effect on the reproduction of P. antipodarum by other authors. Due to the fact that previous studies have shown that this snail is also present in the field at this certain site, it was possible to directly link the results with effects that were observed in the field. This study indicates that effect directed analyses, supported by in vivo biotests, are very useful tools in order to identify the compounds that cause adverse effects on organisms or even population level.
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