title: Preparation and Characterization of Cysteine Adducts of Deoxynivalenol.
abstract: Conjugation with the biologically relevant thiol glutathione is one of the metabolic pathways for the mycotoxin deoxynivalenol (DON) in wheat. The occurrence of putative DON-cysteine conjugates has also been shown in wheat, likely in part as a result of degradation of the glutathione conjugates. It was reported that thiols react in vitro with DON at two positions: reversibly at C-10 of the α,β-unsaturated ketone and irreversibly at C-13 of the epoxy group. We synthesized pure DON-cysteine adducts and made analytical standards using quantitative NMR experiments. Compounds were characterized using NMR and LC-HRMS/MS and tested in vitro for toxicity. Cysteine conjugates were much less toxic than DON at the same concentration, and LC-HRMS analysis demonstrated that there was no detectable metabolism of the conjugates in human monocytes or human macrophages.
authors.affiliation: Norwegian Veterinary Institute , P.O. Box 750, Sentrum, NO-0106 Oslo, Norway.|Department of Chemistry, University of Oslo , P.O. Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Ana
authors.initials: A
authors.lastname: Stanic
authors.affiliation: Norwegian Veterinary Institute , P.O. Box 750, Sentrum, NO-0106 Oslo, Norway.
authors.forename: Silvio
authors.initials: S
authors.lastname: Uhlig
authors.affiliation: Norwegian Veterinary Institute , P.O. Box 750, Sentrum, NO-0106 Oslo, Norway.
authors.forename: Anita
authors.initials: A
authors.lastname: Solhaug
authors.affiliation: Department of Chemistry, University of Oslo , P.O. Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Frode
authors.initials: F
authors.lastname: Rise
authors.affiliation: Norwegian Veterinary Institute , P.O. Box 750, Sentrum, NO-0106 Oslo, Norway.|Chemistry Department, University of Waikato , Private Bag 3105, 3240 Hamilton, New Zealand.
authors.forename: Alistair L
authors.initials: AL
authors.lastname: Wilkins
authors.affiliation: Norwegian Veterinary Institute , P.O. Box 750, Sentrum, NO-0106 Oslo, Norway.
authors.forename: Christopher O
authors.initials: CO
authors.lastname: Miles
mesh_terms: D002460:Cell Line; D002853:Chromatography, Liquid / Q000379:methods; D003470:Culture Media / Q000032:analysis / Q000737:chemistry; D003545:Cysteine / Q000032:analysis* / Q000737:chemistry* / Q000633:toxicity; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D009183:Mycotoxins / Q000032:analysis; D010084:Oxidation-Reduction; D053719:Tandem Mass Spectrometry / Q000379:methods; D014255:Trichothecenes / Q000032:analysis* / Q000737:chemistry* / Q000633:toxicity; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D003470:Culture Media; D009183:Mycotoxins; D014255:Trichothecenes; D014409:Tumor Necrosis Factor-alpha; C007262:deoxynivalenol; D003545:Cysteine
keywords: DON; ERETIC; HRMS; LC−MS; Michael adduct; NMR; qNMR; thiol