Testing Endocrine Disruption in Biota Samples: A Method to Remove Interfering Lipids and Natural Hormones

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).

SIMON Eszter;  LAMOREE Marja;  HAMERS Timo;  WEISS Jana;  BALAAM Jan;  DE BOER Jacob;  LEONARDS Pim; 

2010-11-08

AMER CHEMICAL SOC

JRC59000

0013-936X,   

https://publications.jrc.ec.europa.eu/repository/handle/JRC59000,   

10.1021/es101912z,