Simultaneous determination of aflatoxins, ochratoxin A and Fusarium toxins in maize by liquid chromatography/tandem mass spectrometry after multitoxin immunoaffinity cleanup.

A liquid chromatography tandem mass spectrometry method was developed for the simultaneous determination of aflatoxins (B1, B2, G1, G2), ochratoxin A, fumonisins (B1, B2), deoxynivalenol, zearalenone, T-2 and HT-2 toxins in maize. A double extraction approach, using a phosphate buffered solution followed by methanol, was applied to achieve effective co-extraction of the 11 mycotoxins under investigation having quite different polarity and chemical structure. A new multitoxin immunoaffinity column containing antibodies for all these mycotoxins was used to clean up the extract. Detection and quantification of the 11 mycotoxins were performed by reversed phase liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (HPLC-ESI-MS/MS), using as chromatographic mobile phase a linear gradient of methanol/water containing 0.5% acetic acid and 1 mM ammonium acetate. Method performances were quite satisfactory for all tested mycotoxins at contamination levels close to or below the relevant EU maximum permitted or recommended levels. Limits of detection in maize ranged from 0.4 µg/kg to 5 µg/kg. Recoveries higher than 79% were obtained for all tested mycotoxins with relative standard deviations less than 13%.