Abstract
Diclofenac is a frequently prescribed drug for rheumatic diseases and muscle pain. In rare cases, it may be associated with a severe hepatotoxicity. In literature, it is discussed whether this toxicity is related to the oxidative phase I metabolism, resulting in electrophilic quinone imines, which can subsequently react with nucleophiles present in the liver in form of glutathione or proteins. In this work, electrochemistry coupled to mass spectrometry is used as a tool for the simulation of the oxidative pathway of diclofenac. Using this purely instrumental approach, diclofenac was oxidized in a thin layer cell equipped with a boron doped diamond working electrode. Sum formulae of generated oxidation products were calculated based on accurate mass measurements with deviations below 2 ppm. Quinone imines from diclofenac were detected using this approach. It could be shown for the first time that these quinone imines do not react with glutathione exclusively but also with larger molecules such as the model protein β-lactoglobulin A. A tryptic digest of the generated drug–protein adduct confirms that the protein is modified at the only free thiol-containing peptide. This simple and purely instrumental set-up offers the possibility of generating reactive metabolites of diclofenac and to assess their reactivity rapidly and easily.
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Acknowledgement
The German Federal Environmental Foundation (Deutsche Bundesstiftung Umwelt, DBU, Osnabrück, Germany) is gratefully acknowledged for financial support in form of a Ph.D. scholarship for Helene Faber.
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Published in the special paper collection on Electrochemistry–Mass Spectrometry with guest editors Uwe Karst and Martin Vogel.
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Faber, H., Melles, D., Brauckmann, C. et al. Simulation of the oxidative metabolism of diclofenac by electrochemistry/(liquid chromatography/)mass spectrometry. Anal Bioanal Chem 403, 345–354 (2012). https://doi.org/10.1007/s00216-011-5665-0
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DOI: https://doi.org/10.1007/s00216-011-5665-0