Validation of the 11+Myco MS-PREP® Method for Determination of Aflatoxins, Fumonisins, Deoxynivalenol, Ochratoxin A, Zearalenone, HT-2, and T-2 Toxins in Cereals, Baby Food, Spices, and Animal Feed by Immunoaffinity Column with LC-MS/MS: AOAC Performance Tested MethodSM 112401.

BACKGROUND: The 11+Myco MS-PREP® immunoaffinity column (IAC) contains a gel suspension of monoclonal antibodies specific to the toxins of interest. Following sample extraction, the IAC is used for cleanup and concentration of mycotoxins prior to analysis by LC with tandem mass spectrometry (LC-MS/MS). OBJECTIVE: This study evaluated the IAC with LC-MS/MS method for Performance Tested MethodSM certification for simultaneous determination and confirmation of aflatoxins (AF) B1, B2, G1, G2, and M1; deoxynivalenol (DON), fumonisins B1, B2, and B3; ochratoxin A (OTA); T-2; HT-2; and zearalenone (ZON) from corn, wheat, cereal-based baby food (with and without dairy ingredients), paprika, chili powder, and animal feed. METHODS: A single extraction method using acetonitrile-water (1 + 1, by volume) was used for all matrixes. The method developer validated all matrixes and an independent laboratory verified method performance on corn and animal feed. Data were analyzed for recovery, repeatability precision, LOD, LOQ, confirmation of identity, and method selectivity. RESULTS: Recovery (72-138%) and repeatability (0.46-24%), with the exception of sporadic data points, were within acceptance criteria. LOQ was estimated as AFB1 0.018-0.32 μg/kg, AFB2 0.037-0.28 μg/kg, AFG1 0.019-0.14 μg/kg, AFG2 0.036-0.28 μg/kg, DON 4.0-75 μg/kg, fumonisin B1 4.9-37 μg/kg, fumonisin B2 4.0-32 μg/kg, fumonisin B3 2.0-16 μg/kg, OTA 0.15-4.4 μg/kg, T-2 0.5-7.5 μg/kg, HT-2 0.70-7.5 μg/kg, and ZON 1.3-7.2 μg/kg, depending on matrix. Method performance was verified with reference and QC materials. Selectivity and confirmation of identity were also demonstrated. CONCLUSION: The 11+Myco MS-PREP IAC with LC-MS/MS method demonstrated acceptable performance for simultaneous determination of 12 mycotoxins in seven matrixes. HIGHLIGHT: The data were reviewed by the AOAC Performance Tested MethodsSM Program and approval was granted for certification of the 11+Myco MS-PREP Method as PTM 112401.