Analytical and Bioanalytical Chemistry

, Volume 394, Issue 5, pp 1361–1373

Analysis of glutathione adducts of patulin by means of liquid chromatography (HPLC) with biochemical detection (BCD) and electrospray ionization tandem mass spectrometry (ESI-MS/MS)

  • Nils Helge Schebb
  • Helene Faber
  • Ronald Maul
  • Ferry Heus
  • Jeroen Kool
  • Hubertus Irth
  • Uwe Karst
Original Paper

DOI: 10.1007/s00216-009-2765-1

Cite this article as:
Schebb, N.H., Faber, H., Maul, R. et al. Anal Bioanal Chem (2009) 394: 1361. doi:10.1007/s00216-009-2765-1

Abstract

A novel method for the identification of glutathione/electrophile adducts that are inhibiting glutathione-S-transferase (GST) activity was developed and applied for the analysis of the mycotoxin patulin. The method is based on high-performance liquid chromatography (HPLC) coupled to a continuous-flow enzyme reactor serving as biochemical detector (BCD) in parallel to electrospray mass spectrometric detection (ESI-MS). This HPLC-BCD technique combines a separation step and the detection of the inhibition and is therefore ideally suited for the analysis of the activity of single patulin/glutathione adducts within a complex mixture of adducts. Two out of at least 15 detected patulin–glutathione adducts showed strong GST inhibition. In ESI-MS, the inhibitory active adducts were characterized by [M + H]+ ions with m/z 462.1138 and m/z 741.2011, respectively. They could be identified as a dihydropyranone adduct containing one molecule glutathione and a ketohexanoic acid bearing two glutathione molecules.

Graphical Abstract

OnlineAbstractFigure

Keywords

HPLC-BCD Patulin–glutathione adducts Glutathione-S-transferase inhibition ESI(+)-MS/MS Fragmentation reactions 

Supplementary material

216_2009_2765_MOESM1_ESM.pdf (71 kb)
ESM I (PDF 71.1 kb)

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Nils Helge Schebb
    • 1
  • Helene Faber
    • 1
  • Ronald Maul
    • 2
  • Ferry Heus
    • 3
  • Jeroen Kool
    • 3
  • Hubertus Irth
    • 3
  • Uwe Karst
    • 1
  1. 1.Institut für Anorganische und Analytische ChemieWestfälische Wilhelms-Universität MünsterMünsterGermany
  2. 2.Center for Cardiovascular Research (CCR), Institut für PharmakologieCharité-Universitätsmedizin BerlinBerlinGermany
  3. 3.Department of Chemistry and Pharmaceutical Sciences, Section Analytical Chemistry & Applied SpectroscopyVrije Universiteit AmsterdamAmsterdamThe Netherlands