Title | Women and smokers have elevated urinary F(2)-isoprostane metabolites: a novel extraction and LC-MS methodology. |
Publication Type | Journal Article |
Year of Publication | 2008 |
Authors | Taylor AW, Bruno RS, Traber MG |
Journal | Lipids |
Volume | 43 |
Issue | 10 |
Pagination | 925-36 |
Date Published | 2008 Oct |
ISSN | 0024-4201 |
Keywords | Chromatography, High Pressure Liquid, Dinoprost, F2-Isoprostanes, Female, Humans, Male, Mass Spectrometry, Oxidative Stress, Sex Factors, Smoking, Solid Phase Extraction |
Abstract | 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. |
DOI | 10.1007/s11745-008-3222-1 |
Alternate Journal | Lipids |
PubMed ID | 18751751 |
PubMed Central ID | PMC2770443 |
Grant List | R01DK059576 / 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 |