TitleLC-MS/MS quantitation of mercapturic acid conjugates of lipid peroxidation products as markers of oxidative stress.
Publication TypeJournal Article
Year of Publication2011
AuthorsKuiper HC, Stevens JF
JournalCurr Protoc Toxicol
VolumeChapter 17
Date Published2011 Feb 01
KeywordsAcetylcysteine, Biomarkers, Calibration, Chromatography, Liquid, Humans, Lipid Peroxides, Oxidative Stress, Spectrometry, Mass, Electrospray Ionization, Tandem Mass Spectrometry

Oxidative stress-induced lipid peroxidation (LPO) leads to the formation of cytotoxic and genotoxic 2-alkenals. LPO products such as 4-hydroxy-2(E)-nonenal (HNE) and 4-oxo-2(E)-nonenal (ONE) have been the subject of many studies due to their association with the development of cardiovascular and neurodegenerative diseases, as well as cancer. LPO products are excreted in the urine after conjugation with glutathione (GSH) and subsequent metabolism to mercapturic acid (MA) conjugates. Urinary LPO-MA metabolites are stable end-product metabolites and have gained interest as non-invasive in vivo biomarkers of oxidative stress. This protocol describes a method for the quantitative analysis of LPO-MA metabolites in urine using isotope-dilution liquid chromatography coupled with electrospray tandem mass spectrometry (LC-MS/MS). Included are protocols for preparation of labeled LPO-MA conjugates from unlabeled LPO products and deuterium labeled MA.

Alternate JournalCurr Protoc Toxicol
PubMed ID21442005
PubMed Central IDPMC3062851
Grant ListP30 ES000210-35 / ES / NIEHS NIH HHS / United States
S10 RR022589 / RR / NCRR NIH HHS / United States
P30ES000210 / ES / NIEHS NIH HHS / United States
S10RR022589 / RR / NCRR NIH HHS / United States
R01 HL081721-01A1 / HL / NHLBI NIH HHS / United States
R01 HL081721 / HL / NHLBI NIH HHS / United States
S10 RR022589-01 / RR / NCRR NIH HHS / United States
R01HL081721 / HL / NHLBI NIH HHS / United States
P30 ES000210 / ES / NIEHS NIH HHS / United States