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Women and Smokers Have Elevated Urinary F2-Isoprostane Metabolites: A Novel Extraction and LC–MS Methodology

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Lipids

Abstract

F2-Isoprostanes (F2-IsoPs), regio- and stereoisomers of prostaglandin F2α (PGF2α), and urinary F2-IsoP metabolites including 2,3-dinor-5,6-dihydro-8-iso-PGF2α [2,3-dinor-8-iso-PGF1α (2,3-dinor-F1)] and 2,3 dinor-8-iso-PGF2α (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 PGF2α; 8-iso-PGF2α; 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 F2-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 F2-IsoPs metabolites were identified using SRM. The sum of all urinary F2-IsoP metabolites was 50–100 μg/g creatinine indicating their greater abundance than F2-IsoPs. Women had higher F2-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 F2-IsoPs and their metabolites for the determination of oxidative stress.

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Abbreviations

CE:

Collision energies

F2-IsoPs:

F2-Isoprostanes

GC–MS:

Gas chromatography-mass spectrometry

LC–MS:

Liquid chromatography-mass spectrometry

SPE:

Solid phase extraction

SRM:

Selected reaction monitoring

2,3 dinor-F1:

2,3-dinor-5,6-dihydro-8-iso-PGF2α, or 2,3-dinor-8-iso-PGF1α

2,3 dinor-F2:

2,3-dinor-8-iso-PGF2α

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Acknowledgments

The study authors would like to thank Katie Payne for excellent technical assistance, Dr. Clifford Pereira and Cody Olsen for statistical consultations, and Phenomenex for the donation of SPE cartridges used during method development. The project described was supported by grants to MGT (R01DK059576 and R01DK067930 from the National Institute of Diabetes and Digestive and Kidney Diseases and the Office of Dietary Supplements) and to the Environmental Health Sciences Center at Oregon State University from the National Institute of Environmental Health Sciences (NIH P30 ES00210). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Diabetes and Digestive and Kidney Diseases or the National Institutes of Health.

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Authors and Affiliations

  1. Linus Pauling Institute, Oregon State University, Corvallis, OR, 97331-6512, USA

    Alan W. Taylor & Maret G. Traber

  2. Department of Nutritional Sciences, University of Connecticut, Storrs, CT, 06269-4017, USA

    Richard S. Bruno

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  1. Alan W. Taylor
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  2. Richard S. Bruno
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  3. Maret G. Traber
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Correspondence to Maret G. Traber.

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Taylor, A.W., Bruno, R.S. & Traber, M.G. Women and Smokers Have Elevated Urinary F2-Isoprostane Metabolites: A Novel Extraction and LC–MS Methodology. Lipids 43, 925–936 (2008). https://doi.org/10.1007/s11745-008-3222-1

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  • DOI: https://doi.org/10.1007/s11745-008-3222-1

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