title: 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.
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 (13)C-labelled HNE in biotransformation studies to discriminate metabolites from endogens by liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS). (13)C-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/(13)C-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 (13)C-labelled HNE as a tracer for in vitro metabolism. (13)C-Labelled HNE could also be of benefit for in vivo studies.
authors.affiliation: INRA, UMR1331, Toxalim, Research Center in Food Toxicology, F-31027 Toulouse, France. ijouanin@toulouse.inra.fr
authors.forename: I
authors.initials: I
authors.lastname: Jouanin
authors.forename: M
authors.initials: M
authors.lastname: Baradat
authors.forename: M
authors.initials: M
authors.lastname: Gieules
authors.forename: S
authors.initials: S
authors.lastname: Taché
authors.forename: F H F
authors.initials: FH
authors.lastname: Pierre
authors.forename: F
authors.initials: F
authors.lastname: Guéraud
authors.forename: L
authors.initials: L
authors.lastname: Debrauwer
mesh_terms: D000447:Aldehydes / Q000737:chemistry / Q000493:pharmacokinetics*; D000818:Animals; D001711:Biotransformation; D002247:Carbon Isotopes; D002478:Cells, Cultured; D002853:Chromatography, Liquid / Q000379:methods*; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism; D007553:Isotope Labeling; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D000447:Aldehydes; D002247:Carbon Isotopes; C027576:4-hydroxy-2-nonenal