title: Characterization of Deoxynivalenol-Glutathione Conjugates Using Nuclear Magnetic Resonance Spectroscopy and Liquid Chromatography-High-Resolution Mass Spectrometry.
abstract: Glutathione (GSH) conjugates of the mycotoxin 4-deoxynivalenol (DON), 1, have been detected in plants by LC-MS, but their identities were not confirmed due to a lack of standards. We have synthesized DON-GSH conjugates in alkaline solution. The major products 2 and 5 were isolated and their structures determined by mass spectrometry and NMR spectroscopy as GSH adducts at C-13 and C-10 (via epoxide and Michael addition, respectively) of 1. Other Michael addition products were also tentatively identified by LC-MS. Concentrations of 2 and 5 were determined by quantitative NMR and are suitable for use as quantitative standards for LC-MS studies of plant and animal metabolism of 1. LC-MS showed that in the presence of human glutathione S-transferases of the alpha and mu classes, the reaction of DON and GSH proceeded with a half-life of 17 h, identical with the rate of the uncatalyzed reaction rate, indicating an absence of catalysis.
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: Morten
authors.initials: M
authors.lastname: Sandvik
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: D002851:Chromatography, High Pressure Liquid; D005978:Glutathione / Q000737:chemistry*; D005982:Glutathione Transferase / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D021241:Spectrometry, Mass, Electrospray Ionization; D014255:Trichothecenes / Q000737:chemistry*
chemical_list: D014255:Trichothecenes; D005982:Glutathione Transferase; D005978:Glutathione; C007262:deoxynivalenol
keywords: ERETIC; GSH; GST; HRMS; LC-MS; NMR; conjugation; mycotoxin; thiol