Validation study on urinary biomarkers of exposure for aflatoxin B-1, ochratoxin A, fumonisin B-1, deoxynivalenol and zearalenone in piglets
Résumé
The multi-biomarker approach was used to validate urinary biomarkers in piglets administered boluses contaminated with mixtures of deoxynivalenol (DON), aflatoxin B-1 (AFB(1)), fumonisin B-1 (FB1), zearalenone (ZEA) and ochratoxin A (OTA) at different concentrations. Boluses contaminated with mycotoxins were prepared by slurrying and freeze-drying feed material fortified with culture extracts of selected toxigenic fungi. Piglets were individually placed in metabolic cages to collect urine before gavage and 24 h post dose. Urine samples were hydrolysed with beta-glucuronidase and analysed by a multi-biomarker LC-MS/MS method developed and validated to identify and measure biomarkers of FB1, OTA, DON, ZEA and AFB(1). Urinary levels of FB1, OTA, DON + de-epoxy-deoxynivalenol, ZEA + alpha-zearalenol and aflatoxin M-1 were selected as biomarkers of FB1, OTA, DON, ZEA and AFB(1) ,respectively. Mean percentages of dietary mycotoxins excreted as biomarkers in 24 h post dose urine were 36.8% for ZEA, 28.5% for DON, 2.6% FB1, 2.6% for OTA and 2.5% for AFB(1). A good correlation was observed between the amount of mycotoxins ingested and the amount of relevant biomarkers excreted in 24 h post dose urine. Linear dose-response correlation coefficients ranged between 0.68 and 0.78 for the tested couples of mycotoxin/biomarker. The good sensitivity of the LC-MS/MS method and the good dose-response correlations observed in this study permitted to validate the selected mycotoxin biomarkers in piglets at dietary levels close to the maximum permitted levels reported in Commission Directive 2003/100/EC for AFB(1) and the guidance values reported in Commission Recommendation 2006/576/EC for DON, ZEA, OTA and FB1.
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