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|Title:||Testing Endocrine Disruption in Biota Samples: A Method to Remove Interfering Lipids and Natural Hormones|
|Authors:||SIMON Eszter; LAMOREE Marja; HAMERS Timo; WEISS JANA; BALAAM Jan; DE BOER Jacob; LEONARDS Pim|
|Citation:||ENVIRONMENTAL SCIENCE & TECHNOLOGY vol. 44 no. 21 p. 8322-8329|
|Publisher:||AMER CHEMICAL SOC|
|Type:||Articles in periodicals and books|
|Abstract:||A cleanup method was developed to remove co-extracted lipids and natural hormones from biota samples to enable their testing in in vitro bioassays. Unspiked and spiked fish tissues were extracted and cleaned with a combination of dialysis, gel permeation chromatography (GPC) and normal-phase liquid chromatography (NP-HPLC). Chemical recoveries of the test compounds, thyroid hormone-like and (anti-)androgenic activities of the cleaned extracts were investigated. Despite the chemical and toxicological complexity of the spiking mixture and the sequential sample treatment, chemical analysis revealed acceptable recoveries on avarage: 89±8% after each cleanup step separately and 75±3% after the whole extraction and cleanup. In addition, measured biological activities were in good agreement with the spiking levels. GC-MS lipid profiling revealed the presence of some fatty acids and cholesterol in the final extract, but their influence was negligible at dilution levels required for the bioassays. Co-extracted natural hormones in the fish extracts could be separated from xenobiotics by NP-HPLC fractionation. The developed cleanup method proved to be capable of lipid and natural hormones removal from fish extracts, enabling the application of T4*-TTR and AR-CALUX® bioassays. The method can be used as a sample preparation method of biota samples for toxicity profiling and Effect-Directed Analysis (EDA).|
|JRC Directorate:||Sustainable Resources|
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