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Harmonized Collaborative Validation of a Simultaneous and Multiple Determination Method for Nivalenol, Deoxynivalenol, T-2 Toxin, HT-2 Toxin, and Zearalenone in Wheat and Barley by Liquid Chromatography Coupled to Tandem Mass Spectrometry (LC-MS/MS)

Hiroyuki Nakagawa, Shigehiro Naito, Yuusuke Kitani, Yoshinao Ito, Yoshikazu Aoyama, Matsuhisa Koyama, Yuusuke Hiejima, Keisuke Nakamura, Hiroshi Miyazaki, Horie-Ishiguro Morita, Masayoshi Tamura, Naoki Mochizuki, Masaru Nakamura, Yuusuke Seki, Hisae Kadokura, Hidaka Ikeda, Tomoko Horie-Ishiguro, Yoriko Saito, Miyoko Tajima, Yohsuke Shigemasu, Kikuko

Harmonized collaborative validation of a simultaneous and multiple determination method for nivalenol, deoxynivalenol, T-2 toxin, HT-2 toxin, and zearalenone in wheat and barley by liquid chromatography tandem mass spectrometry (LC-MS/MS) was conducted by participants from 12 laboratories. The fortified samples of wheat and barley at three different levels and one naturally contaminated wheat sample were extracted, consecutively purified through a Presep C18 (ODS) solid phase extraction column and a Bond Elut Mycotoxin multifunctional column, and were analyzed by LC-MS/MS. The employment of internal standards (verrucarol and zearalanone) was apparently effective to ensure repeatability and reproducibility with sufficient recovery of each mycotoxin. This is the first report of the harmonized collaborative validation study of a simultaneous and multiple determination method for both type A and B trichothecenes along with zearalenone by LC-MS/MS. The validated method should be practical for monitoring of the major Fusarium mycotoxins contained in wheat and barley.