Quantitation of plasma total 15-series F(2)-isoprostanes by sequential solid phase and liquid-liquid extraction.
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).
Taylor
Alan W
AW
Linus Pauling Institute, Oregon State University, Corvallis, OR 97331, USA. alan.taylor@oregonstate.edu
Traber
Maret G
MG
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015237:Dinoprost / Q000031:analogs & derivatives / Q000032:analysis; D028441:F2-Isoprostanes / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D015227:Lipid Peroxidation; D052616:Solid Phase Extraction / Q000379:methods*; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*
D028441:F2-Isoprostanes; C075750:8-epi-prostaglandin F2alpha; D015237:Dinoprost