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)
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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.
KeywordsHPLC-BCD Patulin–glutathione adducts Glutathione-S-transferase inhibition ESI(+)-MS/MS Fragmentation reactions
We thank Dr. Heinrich Luftmann (University of Münster, Germany) for helpful discussions. The “Studienstiftung des Deutschen Volkes” (Bonn, Germany) is gratefully acknowledged for financial support in form of a Ph.D. scholarship for Nils Helge Schebb.
- 3.Drusch S, Ragab W (2003) J Food Prot 66:1514–1527Google Scholar
- 4.WHO (1995) World Health Organ Tech Rep Ser 859:1–54Google Scholar
- 14.Dickens F, Jones HEH (1961) Brit J Cancer 15:85–100Google Scholar
- 19.Larsson P, Tjalve H (1992) Cancer Res 52:1267–1277Google Scholar
- 22.Rychlik M (2005) Nutrition 29:61–68Google Scholar
- 24.Lindroth S, von Wright A (1990) J Environ Pathol Toxicol Oncol 10:254–259Google Scholar
- 25.Lindroth S, von Wright A (1978) Appl Environ Microbiol 35:1003–1007Google Scholar
- 32.Lake BG (1987) In: Snell K, Mullock B (eds) Biochemical toxicology: a practical approach. IRL Press, Oxford, pp 183–215Google Scholar
- 39.van Ommen B, Ploemen JH, Bogaards JJ, Monks TJ, Gau SS, van Bladeren PJ (1991) Biochem J 276(Pt 3):661–666Google Scholar
- 41.Ashoor SH, Chu FS (1973) Food Cosmet Toxicol 11:995–1000Google Scholar
- 44.Askelof P, Guthenberg C, Jakobson I, Mannervik B (1975) Biochemical J 147:513–522Google Scholar