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Liquid chromatography/electrospray ionisation mass spectrometric tracking of 4-hydroxy-2(E)-nonenal biotransformations by mouse colon epithelial cells using [1,2-13C2]-4-hydroxy-2(E)-nonenal as stable isotope tracer

Authors


  • Presented at the 6th Congress of the French Society of Stable Isotopes (Société Française des Isotopes Stables, SFIS) held 26–29 October 2010 in Toulouse, France.

I. Jouanin, INRA, UMR1331, Toxalim, Research Center in Food Toxicology, F-31027 Toulouse, France.

E-mail: ijouanin@toulouse.inra.fr

Abstract

4-Hydroxy-2(E)-nonenal (HNE), a product of lipid peroxidation, has been extensively studied in several areas, including metabolism with radio-isotopes and quantification in various matrices with deuterium-labelled HNE as standard. The aim of this work was to evaluate the relevance of 13C-labelled HNE in biotransformation studies to discriminate metabolites from endogens by liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS). 13C-Labelled HNE was synthesised in improved overall yield (20%), with the incorporation of two labels in the molecule. Immortalised mouse colon epithelial cells were incubated with 2:3 molar amounts of HNE/13C-HNE in order to gain information on the detection of metabolites in complex media. Our results demonstrated that the stable isotope m/z values determined by mass spectrometry were relevant in distinguishing metabolites from endogens, and that metabolite structures could be deduced. Six conjugate metabolites and 4-hydroxy-2(E)-nonenoic acid were identified, together with an incompletely identified metabolite. Stable-isotope-labelled HNE has already been used for quantification purposes. However, this is the first report on the use of 13C-labelled HNE as a tracer for in vitro metabolism. 13C-Labelled HNE could also be of benefit for in vivo studies. Copyright © 2011 John Wiley & Sons, Ltd.

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