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Quantitation of plasma total 15-series F(2)-isoprostanes by sequential solid phase and liquid-liquid extraction
Alan W Taylor1, Maret G Traber
1Linus Pauling Institute, Oregon State University, Corvallis, OR 97331, USA. alan.taylor@oregonstate.edu
Abstract:
F(2)-isoprostanes are stereo- and regioisomers of prostaglandin F(2alpha) (PGF(2alpha)) and are used as biomarkers for lipid peroxidation. We modified our liquid-liquid extraction (LLE) procedure for F(2)-isoprostane analysis (Anal. Biochem. 350 (2006) 41-51) to use a combination of solid phase extraction (SPE) and LLE to produce a cleaner extract that can be easily concentrated. In addition, shortening the high-performance liquid chromatography (HPLC) separation increased peak heights in HPLC-tandem mass spectrometry (HPLC-MS/MS) analysis. Both changes increased the overall sensitivity of the assay. MS/MS analysis served to confirm the identity of specific peaks that may be better biomarkers than the commonly measured 8-iso-PGF(2alpha).
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