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Year :	2007
Tome :	158
Volume :	10
Pages :	504-508
Title :	HPLC assay of zearalenone and reduced metabolites in S9 fractions of duck liver
Authors :	M. KOLF-CLAUW, F. AYOUNI, D. TARDIEU, P. GUERRE
Summary :	HPLC analysis of zearalenone (ZEA), zearalenols (?-ZOL and ß-ZOL) and zearalanols (?-ZAL and ß-ZAL) was developed, in order to obtain a sensitive and reproducible method to quantify ZEA and its reduced metabolites in subcellular fractions of animal livers (S9 samples). Optimal in vitro metabolism was observed by incubating 5 mg S9 proteins with 0.016 ?mol. ZEA. Acetonitrile and diethylether/chloroform mixture were compared for extraction, as well as different mobile phases and two detection modes in HPLC analysis. Extracted samples were eluted with water/acetonitrile (55:45, v/v) at a flow-rate of 1.0 ml/min-1, resulting in well separated peaks between ZEA and the metabolites. The limits of detection ranged from 0.5 to 2 ng/mg S9 proteins using UV, and from 0.04 to 4 ng/mg S9 proteins, using fluorescence detection. Fluorescence showed a ten-fold higher sensitivity than UV detection for ZEA and ?-ZOL. Repeatability (10 assays) was 2.7% to 6.99% for zearalenols. Day-by-day coefficients of variation for zearalenone and zeranols with UV detection were 3.3 to 8.5 %, and 2.5 to 4.3 %, respectively. This analysis applied to S9 samples from ducks after 30 min of ZEA incubation allowed to demonstrate that ?-ZOL is the main reduced metabolite in the duck. The present method is particularly adapted for studying in vitro metabolism of ZEA and inter-species variations.
Keywords :	Zearalenone, zearalenols, S9 liver fractions, duck, metabolism, HPLC.
Correspondence :	M. KOLF-CLAUW
Adress :	Unité de Mycotoxicologie, Ecole Nationale Vétérinaire de Toulouse, 23 chemin des Capelles, BP 87214, 31076 Toulouse Cedex FRANCE - [email protected]
Link :	pdf

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