Sample preparation method for the ER-CALUX bioassay screening of (xeno-)estrogenic activity in sediment extracts

C.J. Houtman, P.E.G. Leonards, W. Kapiteijn, J.F. Bakker, A. Brouwer, M.H. Lamoree, J. Legler, H.J.C. Klamer

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The application of bioassays to assess the occurrence of estrogenic compounds in the environment is increasing in both a scientific and statutory context. The availability of appropriate validated methods for sample pre-treatment and analysis is crucial for the successful implementation of bioassays. Here, we present a sample preparation method for the bioassay screening of estrogenic activity in sediment with the in vitro Estrogen Receptor mediated Chemical Activated LUciferase gene eXpression (ER-CALUX®) assay. The method makes use of an Accelerated Solvent (ASE) or Soxhlet extraction with a mixture of dichloromethane and acetone (3:1, v/v), followed by clean up of the extract by Gel Permeation Chromatography (GPC). Recoveries of a panel of 17 pollutants differing largely in physical-chemical properties from spiked sediment were determined and appeared to be on average about 86%. Furthermore, the estrogenic potencies of all test compounds were individually assessed by determination of concentration-response relationships in the ER-CALUX assay. Concentration dependent estrogenic potency was found for 14 of the 17 compounds, with potencies of about 10
Original languageEnglish
Pages (from-to)134-144
Number of pages12
JournalScience of the Total Environment
Publication statusPublished - 2007


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