title: Enzyme immunoassay for a urinary metabolite of 4-hydroxynonenal as a marker of lipid peroxidation.
abstract: Free radical reactions are involved in the pathogenesis of numerous diseases, so there is a real need to develop biomarkers that reflect these reactions in vivo. 4-Hydroxy-2-nonenal (HNE) is a major product of the lipid peroxidation process that is a consequence of free radical reactions. We present here the development and validation of an enzyme immunoassay (EIA) of the major urinary metabolite of HNE, namely 1,4-dihydroxynonane-mercapturic acid (DHN-MA). EIA allowed direct measurement of DHN-MA in rat urine with good sensitivity (0.02 ng/ml) and precision (intraassay CV = 5.7%). Recovery was complete (99-102%). Cross-reactivity was very low with 1,4-dihydroxynonene and with different mercapturic acids except with one other HNE urinary metabolite. Good correlation (EIA = 0.79 x LC/MS + 14.03, r = 0.877, p < 10(-8)) was obtained between EIA and liquid chromatography/mass spectrometry (LC/MS) quantitation when analyzing urine samples of rats with different oxidative status, due to treatment with either BrCCl(3) or trinitrobenzene sulfonic acid, which are known to induce hepatic lipid peroxidation or colon inflammation, respectively.
authors.affiliation: Institut National de la Recherche Agronomique, UMR-1089 Xénobiotiques, INRA/ENVT, BP 3, 180 chemin de Tournefeuille, 31931 Toulouse Cedex 9, France. fgueraud@toulouse.inra.fr
authors.forename: Françoise
authors.initials: F
authors.lastname: Guéraud
authors.forename: Géraldine
authors.initials: G
authors.lastname: Peiro
authors.forename: Hervé
authors.initials: H
authors.lastname: Bernard
authors.forename: Jacques
authors.initials: J
authors.lastname: Alary
authors.forename: Christophe
authors.initials: C
authors.lastname: Créminon
authors.forename: Laurent
authors.initials: L
authors.lastname: Debrauwer
authors.forename: Estelle
authors.initials: E
authors.lastname: Rathahao
authors.forename: Marie-Françoise
authors.initials: MF
authors.lastname: Drumare
authors.forename: Cécile
authors.initials: C
authors.lastname: Canlet
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Wal
authors.forename: Georges
authors.initials: G
authors.lastname: Bories
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000276:immunology / Q000652:urine; D000447:Aldehydes / Q000652:urine*; D000475:Alkenes / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000652:urine*; D001975:Bromotrichloromethane / Q000494:pharmacology; D002853:Chromatography, Liquid; D003429:Cross Reactions; D005609:Free Radicals; D007124:Immunoenzyme Techniques; D015227:Lipid Peroxidation; D008297:Male; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology
chemical_list: C091765:1,4-dihydroxy-2-nonene; C501116:1,4-dihydroxynonene mercapturic acid; D000447:Aldehydes; D000475:Alkenes; D015415:Biomarkers; D005609:Free Radicals; D014302:Trinitrobenzenesulfonic Acid; D001975:Bromotrichloromethane; C027576:4-hydroxy-2-nonenal; D000111:Acetylcysteine