Analysis of F2-isoprostanes in plasma of pregnant women by HPLC-MS/MS using a column packed with core-shell particles

Plasma F-2 -isoprostanes (F-2-isoPs) are reliable bio-markers of oxidative stress. Several possible F-2-isoPs are generated by the oxidation of arachidonic acid esterified in phospholipids. The separation of these isomers represents a technical challenge for rapid and selective determination. We have developed a HPLC-MS/MS method for the simultaneous determination of seven plasma F-2-isoPs, namely 8-iso-15(R)-prostaglandin F-2 alpha (PGF(2 alpha)), 8-iso-PGF(2 alpha), 15(R)-PGF(2 alpha), iPF(2 alpha)-IV, iPF(2 alpha)-VI, 5-iPF(2 alpha)-VI, and (+/-) 5-8,12-iso-iPF(2 alpha)-VI. We have validated this method in plasma of pregnant women, a mild physiological oxidative stress known to increase F-2-isoPs. Thus, plasma samples of women collected at the third trimester of pregnancy (n = 20) were subjected to alkaline hydrolysis followed by liquid-liquid extraction in order to extract total F-2-isoPs. The F-2-isoPs were separated within 16.5 min using a column packed with core-shell particles. The class VI isomers were the most abundant, accounting for 65% of the total level of all quantified F-2-isoPs in plasma of pregnant women (P < 0.05). The 15(R)-PGF(2 alpha) was the most abundant of the class III isomers quantified. This method allowed fast and selective separation of seven isomers from three different classes of F-2-isoP regioisomers.-Larose, J., P. Julien, and J.-F. Bilodeau. Analysis of F-2-isoprostanes in plasma of pregnant women by HPLC-MS/MS using a column packed with core-shell particles. J. Lipid Res. 2013. 54: 1505-1511.