Quantitative determination and structure elucidation of type A- and B-trichothecenes by HPLC/ion trap multiple mass spectrometry

A method is presented for the quantification and structure confirmation of trichothecenes in wheat by high-performance liquid chromatography combined with multiple mass spectrometry (MSn). Nine type A- and B-trichothecenes we re determined (nivalenol, deoxynivalenol, fusarenon-X, 3-acetyldeoxynivalen ol, 15-acetyldeoxynitralenol, neosolaniol, diacetoxyscirpenol, HT-2 toxin, and T-2 toxin). Extraction was carried out with acetonitrile/water. The ext ract was purified on a MycoSep column. Quantification was based on an inter nal standard and atmospheric pressure chemical ionization in the positive i on mode. Recoveries from spiked wheat were in the range of 80-106% at level s of 500 ppb. The limits of quantification for the whole method were betwee n 10 and 100 ppb. Ion adduct formation with ammonium and acetate ions and M Sn experiments provided information about substitution and fragmentation be havior of the mycotoxins. A scheme has been established for the partial str ucture elucidation of type A- and B-trichothecenes in fungal cultures.