HPLC method validation for simultaneous determination of three mycotoxins from corn seeds

Abstract

A new HPLC optimized method for simultaneous determination of some mycotoxins by separation and using a Dionex UltiMate3000 modular system, with multichannel UV detector was developed and validated. The method optimization was performed to determine simultaneously, several relevant mycotoxins from corn seeds that were stored over 8 months. The mycotoxins selected for quantification were: aflatoxin B1, ochratoxin A and zearalenone. They were selected due to their high concentration in the fodder. For the development stages of the HPLC method, it was introduced an internal standard to have accurate results. Taking into account that the analytes should be extracted from the complex matrix they reside, an extraction procedure was performed, using organic solvents, and the selection after repeated tests demonstrated the best capacity for ethyl acetate. The recovery is about 60-70 % after the extraction process also there is a good preconcentration (×2.5) of the analytes and the internal standard for their quantification. The limit of quantification (LOQ) obtained by chromatographic parameters optimization, for aflatoxin B1 and ochratoxin A were about 3-5 μg kg -1 and 14.4 μg μg kg -1 for zearalenone of raw biological material, making these values lower than those accepted by the actual normatives and regulations.