Simultaneous Determination of Aflatoxins, Ochratoxin A, and Zearalenone in Grains by New Immunoaffinity Column/Liquid Chromatography

Abstract

Abstract The simultaneous determination of mycotoxins was performed in 3 steps: extraction, cleanup, and detection. For extraction, a mixture of acetonitrile–water (60 + 40, v/v) was proved appropriate. For cleanup, a new Afla-Ochra-Zea immunoaffinity column was used. After derivatization with trifluoroacetic acid, the mycotoxins aflatoxins, ochratoxin A (OTA), and zearalenone (ZEA) were determined simultaneously by liquid chromatography with fluorescence detection. The detection limits in different matrixes after cleanup with the new immunoaffinity column were very low: aflatoxins, 0.002–0.7 μg/kg; OTA, 0.07–0.25 μg/kg; ZEA, 1–3 μg/kg. The limits of determination were: aflatoxins, 0.25 μg/kg; OTA, 0.5 μg/kg; ZEA, 5 μg/kg. The recovery rates for aflatoxins, OTA, and ZEA for rye and rice were between 86 and 93% when a 0.5 g sample matter per immunoaffinity column was used.