Analysis of Deoxynivalenol and De-Epoxy- Deoxynivalenol in Horse Blood Through Liquid Chromatography After Clean-up with Immunoaffinifty Column

F. M. C. S. Setyabudi, J. Böhm , H. K. Mayer , F. E. Razazzi

Abstract

The Fusarium toxin, deoxynivalenol (DON) has a great significance in human and animal nutrition. Five Haflinger mares were selected to study the concentration of Deoxynivalenol (DON) and De-Epoxy-Deoxynivalenol (deepoxy-DON) in plasma samples of horse fed a diet contaminated with DON. The experimental animals were feed contaminated oat (12mg /kg oat), total being 36 mg DON/day. A series of blood samples were carried out on the day of feeding (day 1) and day 10. The concentration of DON and deepoxy-DON were determined using liquid chromatographic (HPLC-UV) method coupled with the immunoaffinity columns. The method was validated with the range of 10-100 ng/mL DON and deepoxy-DON. The Relative Standard Deviation (RSD) intra-day and inter-day for DON were less than 9% and 7% and for deepoxy-DON were less than 11% and 9%, respectively. The average recoveries of both analytes were found to be approximately 81.9 ± 5.3 % and 80.5 ± 6.4%, respectively. Generally, free form of DON (without treatment with ß-glucuronidase) in the plasma samples collected 24 hour post-feeding on day ten were under the limit of detection in all animals except in two animals (11.7 and 7.6 ng/mL). The higher concentration of DON was found in the plasma one hour post-feeding. In plasma, it seems that majority of DON is present in the glucuronide-conjugated form. The levels detected are not very high, but even long-term exposure to low doses of these mycotoxins may represent a threat to horse health.

Key Words: DON, Horse, HPLC, Mycotoxins, De-Epoxy-Deoxynivalenol