Single laboratory-validated HPLC methods for determination of ochratoxin A, fumonisin B1 and B2, zearalenone and deoxynivalenol in cereals and cereal-based foods

The aim of this study was to optimize analytical methods for determination of ochratoxin A (OTA), fumonisins B1 and B2 (FB1, FB2), zearalenone (ZON), and deoxynivalenol (DON) in cereals and cereal-based food products. The chromatographic separation was performed by reversed phase high performance liquid chromatography (HPLC) with fluorescence detection (FLD) and ultraviolet-diode array detection (UV-DAD). Optimized sample preparation methods utilizing extraction with solvent and clean-up on immunoaffinity column (IAC) were used. The procedures were validated in accordance with single laboratory validation principles. The limit of detection (LOD) for OTA, FB1, FB2 and ZON was 0.081, 50, 40 and 12 mu g.kg(-1), respectively, whereas the limit of quantification (LOQ) for DON was 45 mu g.kg(-1) for processed cereals (group A) and 85 mu g.kg(-1) for unprocessed cereals (group B). Average recoveries varied, in the range of 92-94% for OTA, 93-87% for FB1, 83-96% for FB2, 89-103% for ZON, and 97-87% for DON at defined spiking levels. The accuracy was additionally estimated for OTA, ZON and fumonisins using a certified reference material (CRM). All methods showed good sensitivity, accuracy and precision comparable to those published recently. The methods were successfully applied to the determination of mycotoxins in cereals at examination of occurrence and content of these toxins.