Analysis of deoxynivalenol and de-epoxy-deoxynivalenol in animal tissues by liquid chromatography after clean-up with an immunoaffinity column

A method for the determination of deoxynivalenol (DON) and its metabolite de-epoxy-deoxynivalenol (de-epoxy-DON) in blood serum, urine, faeces/digesta and bile fluid is described. Liquid samples (urine and bile after incubation with ß-glucuronidase) were extracted with ethyl acetate on a ChemElut column. Freeze dried faeces and digesta were extracted with a mixture of acetonitrile and water. For clean-up, serum and urine extracts could be directly applied to an immunoaffinity column (IAC). A pre-treatment prior to the IAC clean-up was necessary in the case of bile and faeces/digesta. DON and de-epoxy-DON were determined by high performance liquid chromatography (HPLC) with diode-array detection (DAD).Because of the clean extracts, low detection limits in the range of 4 ng/ml (serum) to 20 ng/g (dried faeces/digesta) were achieved. The recovery of DON and de-epoxy-DON was in the range of 75-89% and 64-85%, respectively. DON was found in blood serum, urine, bile, digesta and faeces of pigs from several feeding studies with DON contaminated feed; de-epoxy-DON could be detected only in digesta, faeces and urine.