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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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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.

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Acknowledgements

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.

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Authors and Affiliations

  1. Institut für Anorganische und Analytische Chemie, Westfälische Wilhelms-Universität Münster, Corrensstraße 30, 48149, Münster, Germany

    Nils Helge Schebb, Helene Faber & Uwe Karst

  2. Center for Cardiovascular Research (CCR), Institut für Pharmakologie, Charité-Universitätsmedizin Berlin, Hessische Straße 3-4, 10115, Berlin, Germany

    Ronald Maul

  3. Department of Chemistry and Pharmaceutical Sciences, Section Analytical Chemistry & Applied Spectroscopy, Vrije Universiteit Amsterdam, De Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands

    Ferry Heus, Jeroen Kool & Hubertus Irth

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  1. Nils Helge Schebb
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  2. Helene Faber
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  3. Ronald Maul
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  4. Ferry Heus
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  5. Jeroen Kool
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  7. Uwe Karst
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Correspondence to Uwe Karst.

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Schebb, N.H., Faber, H., Maul, R. et al. 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). Anal Bioanal Chem 394, 1361–1373 (2009). https://doi.org/10.1007/s00216-009-2765-1

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  • DOI: https://doi.org/10.1007/s00216-009-2765-1

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