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Women and Smokers Have Elevated Urinary F 2 ‐Isoprostane Metabolites: A Novel Extraction and LC–MS Methodology
Author(s) -
Taylor Alan W.,
Bruno Richard S.,
Traber Maret G.
Publication year - 2008
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-008-3222-1
Subject(s) - metabolite , chemistry , urine , chromatography , creatinine , urinary system , solid phase extraction , isoprostanes , isoprostane , population , oxidative stress , high performance liquid chromatography , medicine , biochemistry , lipid peroxidation , environmental health
F 2 ‐Isoprostanes (F 2 ‐IsoPs), regio‐ and stereoisomers of prostaglandin F 2α (PGF 2α ), and urinary F 2 ‐IsoP metabolites including 2,3‐dinor‐5,6‐dihydro‐8‐ iso ‐PGF 2α [2,3‐dinor‐8‐ iso ‐PGF 1α (2,3‐dinor‐F1)] and 2,3 dinor‐8‐ iso ‐PGF 2α (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 2α ; 8‐ iso ‐PGF 2α ; 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 ~0.2–1.5 μg/g creatinine, 2,3‐dinor‐F1 was ~1–3 μg/g and 2,3‐dinor‐F2 was ~3–5 μg/g. Additional F 2 ‐IsoPs metabolites were identified using SRM. The sum of all urinary F 2 ‐IsoP metabolites was 50–100 μg/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.