Characterization of (13C24) T-2 toxin and its use as an internal standard for the quantification of T-2 toxin in cereals with HPLC–MS/MS

Abstract

In this paper, the structure and the identity of fully 13C-substituted T-2 toxin were confirmed using high-resolution mass spectrometry, 1H-NMR, 13C-NMR, tandem mass spectrometry and HPLC-DAD. The purity of this compound was estimated to be at least 98.8% according to UV data. The isotopic distribution of (13C(24)) T-2 toxin indicated a total isotopic enrichment of 98.2 +/- 1.0 atom% 13C, and the application of different MS measurement modes revealed the MS/MS fragmentation pattern of T-2 toxin. Furthermore, a stable isotope dilution mass spectrometry method for the quantification of T-2 toxin was developed using (13C(24)) T-2 toxin as internal standard. The method was evaluated with and without conventional clean-up and validated for maize and oats. Both cereals showed strong matrix enhancement effects, which could be compensated for through the application of the isotope-substituted internal standard.

Analysis of Fusarium toxins via HPLC-MS/MS multimethods: matrix effects and strategies for compensation

“…Without the use of the IS 13 C 24 -T-2, they observed enhanced toxin signals up to 146% (without cleanup) and 115% (with cleanup) (Haeubl et al 2007). This reversed matrix effect is probably caused by the very short retention time of 3.8 min compared to the retention time observed with the presented DSPE method at 26.3 min.…”

“…The first is more elaborate as every single sample has to be spiked on several concentration levels. Isotope-labeled IS added to the solid sample prior to cleanup can compensate-in an ideal case-both the losses induced by extraction, cleanup and HPLC analysis as stated by several research teams Rychlik 2006, 2007;Beyer et al 2009;Bretz et al 2006;Haeubl et al 2006Haeubl et al , 2007. However, using isotope-labeled IS for every analyte from the beginning of the sample preparation results in very high costs which are not suitable for routine analysis as well.…”

“…The matrix calibration curve showed a 50% lower slope compared to the solvent calibration when the analyte eluted very early (Matuszewski et al 1998). Moreover, Haeubl et al (2006Haeubl et al ( , 2007 used an APCI source, instead of an ESI source which was used in this work. Since the solvent evaporation and the ionization of the analytes are separated steps at the APCI source, different effects regarding matrix interactions can be observed, which could also be the reason for the reversed matrix effects (Haeubl et al 2006(Haeubl et al , 2007Matuszewski et al 1998).…”

“…Previously published methods for the determination of trichothecenes, fumonisins and/or zearalenone either did not include the compensation of such effects (Berthiller et al 2005;Cavaliere et al 2005) or included matrix calibration (Biselli et al 2005;Spanjer et al 2008) which is not feasible for routine work. Some other methods describe the use of one internal standard (IS) for several analytes (Berger et al 1999;Kloetzel et al 2006 (European Commission 1999, 2000a, b, 2002 often describe the use of isotope-labeled IS for compensation of matrix suppression or enhancement Rychlik 2006, 2007;Haeubl et al 2006Haeubl et al , 2007Herebian et al 2009;Rychlik and Asam 2008;Bretz et al 2006;Cramer et al 2007), which is in general an effective method, but very expensive as spiking is commonly performed before extraction. Some of these studies reveal that significantly lower values were obtained when no isotope-labeled IS was used (Haeubl et al 2006;Asam and Rychlik 2007), however, there is only less data about the part of matrix effects which is only caused by ionization.…”

Large-scale production of selected type A trichothecenes: the use of HT-2 toxin and T-2 triol as precursors for the synthesis of d 3-T-2 and d 3-HT-2 toxin

“…In this way, radiolabeled T-2 was prepared by biosynthetic production of F. tricinctum on rice, whereby 14 C-acetate, 14 Cmevalonate and 3 H-mevalonate were added to the culture media (Hagler et al 1981). Fully 13 C-labeled T-2 (Haubl et al 2007) and HT-2, the only isotope labeled standards of T-2 and HT-2 which are commercially available, were also produced biosynthetically.…”