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Title:   Author  item
                                            REITERER, JACQUELINE - University Of Natural Resources & Applied Life Sciences - Austriaitem
                                            VARGA, ELISABETH - University Of Natural Resources & Applied Life Sciences - Austriaitem
                                            NATHANAIL, ALEXIS - Finnish Food Safety Authorityitem
                                            BUESCHL, CHRISTOPH - University Of Natural Resources & Applied Life Sciences - Austriaitem
                                            RECHTHALER, JUSTYNA - University Of Natural Resources & Applied Life Sciences - Austriaitem
                                            McCormick, Susanitem
                                            ADAM, GERHARD - University Of Natural Resources & Applied Life Sciences - Austriaitem
                                            BERTHILLER, FRANZ - University Of Natural Resources & Applied Life Sciences - Austriaitem
                                            LEMMENS, MARC - University Of Natural Resources & Applied Life Sciences - Austriaitem
                                            SCHUHMACHER, RAINER - University Of Natural Resources & Applied Life Sciences - Austria
  Submitted to: Analytical and Bioanalytical Chemistry
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 8/12/2015
                                    Publication  Date: 10/3/2015
                                    Citation: Meng-Reiterer, J., Varga, E., Nathanail, A.V., Bueschl, C., Rechthaler, J., McCormick, S.P., Michlmayr, H., Malachova, A., Fruhmann, P., Adam, G., Berthiller, F., Lemmens, M., Schuhmacher, R. 2015. Tracing the metabolism of HT-2 toxin and T-2 toxin in barley by isotope-assisted untargeted screening and quantitative LC-HRMS analysis. Analytical and Bioanalytical Chemistry. 407(26):8019-8033.Interpretive Summary: T-2 toxin is a trichothecene mycotoxin produced when the fungus Fusarium infects small grains including barley. Ingestion of T-2 toxin contaminated grain can cause feed refusal, diarrhea, and hemorrhage. Plants infected with mycotoxin-producing fungi form sugar derivatives of toxins. These sugar derivatives, sometimes called masked mycotoxins, are a potential food safety concern because they are not detected by standard approaches and could potentially be converted back to the parent toxin during digestion or food processing. This study used sophisticated mass spectrometry methods to identify and track T-2 toxin and twenty-two T-2 toxin related metabolites in barley. Development of accurate methods to assess the occurrence of these metabolites in cereals are needed to fully determine the risk posed by consuming foods contaminated with T-2 toxin. This study found that T-2 toxin and T-2 toxin glucoside are rapidly converted into other metabolites that should be considered when determining the risk of parent toxin.
                                Technical Abstract: An extensive study of the metabolism of the type-A trichothecene mycotoxins HT-2 toxin and T-2 toxin in barley using liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) is reported. A recently developed untargeted approach based on stable isotopic labelling, LC-Orbitrap-MS analysis with fast polarity switching and data processing by MetExtract software was combined with targeted LC-Q-TOF-MS(/MS) analysis for metabolite structure elucidation and quantification. In total, 9 HT-2 toxin and 13 T-2 toxin metabolites plus tentative isomers were detected and were successfully annotated by calculation of elemental formulas, further LC-HRMS/MS measurements, and comparison with authentic standards. As a result, glucosylated forms of the toxins, as well as malonylglucoside, acetyl- and feruloyl-conjugates were elucidated. Additionally, time courses of metabolite formation and mass balances were established. For absolute quantification, the method was validated in-house for those compounds for which standards were available by determining apparent recovery, signal suppression or enhancement and extraction recovery. Interestingly, T-2 toxin was rapidly metabolised to HT-2 toxin while HT-2 toxin-3-O-ß-glucoside was identified (confirmed by authentic standard) as a main metabolite of both parent toxins which reached its maximum concentration one day after toxin treatment.


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