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Quantification of F2-isoprostanes as a biomarker of oxidative stress

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Abstract

Oxidant stress has been implicated in a wide variety of disease processes. One method to quantify oxidative injury is to measure lipid peroxidation. Quantification of a group of prostaglandin F2α-like compounds derived from the nonezymatic oxidation of arachidonic acid, termed the F2-isoprostanes (F2-IsoPs), provides an accurate assessment of oxidative stress both in vitro and in vivo. In fact, in a recent independent study sponsored by the National Institutes of Health (NIH), F2-IsoPs were shown to be the most reliable index of in vivo oxidant stress when compared against other well known biomarkers. This protocol details our laboratory's method to quantify F2-IsoPs in biological fluids and tissues using gas chromatography-mass spectrometry (GC-MS). This procedure can be completed for 12–15 samples in 6–8 h.

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Figure 1
Figure 2
Figure 3: Thin layer chromatography analysis of the methyl ester of PGF2α, the TLC standard, visualized with phosphomolybdic acid.
Figure 4: Analysis of F2-IsoPs in human plasma.

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Acknowledgements

Supported by NIH grants DK48831, CA77839, GM154312 and ES13125.

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Authors and Affiliations

  1. Division of Clinical Pharmacology, Vanderbilt University School of Medicine, Nashville, 37232-6602, Tennessee, USA

    Ginger L Milne, Stephanie C Sanchez, Erik S Musiek & Jason D Morrow

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  1. Ginger L Milne
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  2. Stephanie C Sanchez
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  3. Erik S Musiek
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  4. Jason D Morrow
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Correspondence to Jason D Morrow.

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Milne, G., Sanchez, S., Musiek, E. et al. Quantification of F2-isoprostanes as a biomarker of oxidative stress. Nat Protoc 2, 221–226 (2007). https://doi.org/10.1038/nprot.2006.375

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