TitleWomen and smokers have elevated urinary F(2)-isoprostane metabolites: a novel extraction and LC-MS methodology.
Publication TypeJournal Article
Year of Publication2008
AuthorsTaylor AW, Bruno RS, Traber MG
Date Published2008 Oct
KeywordsChromatography, High Pressure Liquid, Dinoprost, F2-Isoprostanes, Female, Humans, Male, Mass Spectrometry, Oxidative Stress, Sex Factors, Smoking, Solid Phase Extraction

F(2)-Isoprostanes (F(2)-IsoPs), regio- and stereoisomers of prostaglandin F(2alpha) (PGF(2alpha)), and urinary F(2)-IsoP metabolites including 2,3-dinor-5,6-dihydro-8-iso-PGF(2alpha) [2,3-dinor-8-iso-PGF(1alpha) (2,3-dinor-F1)] and 2,3 dinor-8-iso-PGF(2alpha) (2,3-dinor-F2), have all been used as biomarkers of oxidative stress. A novel method was developed to measure these biomarkers using a single solid phase extraction (SPE) cartridge, separation by HPLC, and detection by negative mode selected reaction monitoring (SRM) mass spectrometry (MS), using authentic standards of PGF(2alpha); 8-iso-PGF(2alpha); 2,3-dinor-F1 and 2,3-dinor-F2 to identify specific chromatographic peaks. The method was validated in a population of healthy, college-aged nonsmokers (n = 6 M/8F) and smokers (n = 6 M/5F). Urinary F(2)-IsoP concentrations were approximately 0.2-1.5 microg/g creatinine, 2,3-dinor-F1 was approximately1-3 microg/g and 2,3-dinor-F2 was approximately 3-5 microg/g. Additional F(2)-IsoPs metabolites were identified using SRM. The sum of all urinary F(2)-IsoP metabolites was 50-100 microg/g creatinine indicating their greater abundance than F(2)-IsoPs. Women had higher F(2)-IsoP metabolite concentrations than did men (MANOVA, main effect P = 0.003); cigarette smokers had higher concentrations than did nonsmokers (main effect P = 0.036). For men or women, respectively, smokers had higher metabolite concentrations than did nonsmokers (P < 0.05). Thus, our method simultaneously allows measurement of urinary F(2)-IsoPs and their metabolites for the determination of oxidative stress.

Alternate JournalLipids
PubMed ID18751751
PubMed Central IDPMC2770443
Grant ListR01DK059576 / DK / NIDDK NIH HHS / United States
P30 ES00210 / ES / NIEHS NIH HHS / United States
R01 DK067930 / DK / NIDDK NIH HHS / United States
R01DK067930 / DK / NIDDK NIH HHS / United States
R01 DK067930-01A1S1 / DK / NIDDK NIH HHS / United States
R01 DK081761 / DK / NIDDK NIH HHS / United States
R01 DK059576 / DK / NIDDK NIH HHS / United States
P30 ES000210 / ES / NIEHS NIH HHS / United States