Enzyme immunoassay for a urinary metabolite of 4-hydroxynonenal as a marker of lipid peroxidation.
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.
Guéraud
Françoise
F
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
Peiro
Géraldine
G
Bernard
Hervé
H
Alary
Jacques
J
Créminon
Christophe
C
Debrauwer
Laurent
L
Rathahao
Estelle
E
Drumare
Marie-Françoise
MF
Canlet
Cécile
C
Wal
Jean-Michel
JM
Bories
Georges
G
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
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