Determination of 12 Type A and B Trichothecenes in Cereals by Liquid Chromatography−Electrospray Ionization Tandem Mass Spectrometry

Abstract

A new sensitive method for the simultaneous determination of 12 trichothecenes (deoxynivalenol, nivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, fusarenon X, T-2 toxin, HT-2 toxin, neosolaniol, monoacetoxyscirpenol, diacetoxyscirpenol, T-2 triol, and T-2 tetraol) by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) is presented. The development of the method and investigations on the matrix influence on the MS signal are described in particular. The matrix effect was thereby minimized by using an internal standard, a special mobile phase, and specific fragmentation parameters. The sample was extracted with acetonitrile/water (84:16, v/v), and the extract was cleaned up with a MycoSep 227 column. Quantification was based on the internal standard de-epoxy-deoxynivalenol. Calibration curves were linear between 16 and 1600 ng/g, and the limits of detection ranged from 0.18 to 5.0 ng/g. The developed method was applied for the determination of trichothecenes in 120 naturally contaminated wheat and oat samples.

Stable isotope dilution analysis of the Fusarium mycotoxins deoxynivalenol and 3‐acetyldeoxynivalenol

“…Approaching from HPLC-MS, we compared the response of DON with ESI in both polarities and found it to be more sensitive in the negative mode, in accordance with literature [10,12,14,20,21]. In the course of tuning the instrument to develop an MRM method for quantitation purposes, we recorded the product ion spectrum of [DON-H] -.…”

“…To assure, nevertheless, high accuracy and reproducibility of the results, matrix effects have to be compensated either by laborious matrix calibration [10] or by the use of suitable internal standards (IS). In literature, structural analogues and trichothecenes such as neosolaniol [11], verrucarol [12,13] or deepoxy-deoxynivalenol [14] are described for this purpose. However, the transferability of these IS might be limited, due to differences between analytes and IS in the extraction and ionization efficiency, the recovery, the retention time, etc.…”

No carry over of unmetabolised deoxynivalenol in milk of dairy cows fed high concentrate proportions

“…The LC-ESI-MS/MS method will be described in detail elsewhere (Valenta et al, in preparation). Briefly, the separation was carried out on a Betasil Phenyl/Hexyl column (100 mm62.1 mm, 3 lm; Thermo Electron Corporation, Runcorn, UK), using a binary gradient of 0.13 mM ammonium acetate in water (pH 7.4, solvent A; described by Klötzel et al [31], and ACN (solvent B). The measurements were performed with multiple reaction monitoring (MRM) in the negative mode, selecting the mass transitions 295 265 (DON) and 279 249 (DOM-1) for quantification and 295 138 (DON) and 279 231 (DOM-1) for additional qualifying.…”

Synthesis of stable isotope labeled 3-acetyldeoxynivalenol

“…To assure high accuracy and reproducibility of the quantitative results, either performing laborious matrix calibration [6] or using suitable internal standards is mandatory. Currently, structural analogs of the trichothecenes such as neosolaniol [7], verrucarol [8,9] or deepoxy-DON [10] are common as internal standards. However, stable isotope labeled analytes would be the ideal standards for mass spectrometric detection, as they compensate matrix suppression in HPLC-MS analysis and have a relative response factor close to 1.…”