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. 2013 May;54(5):1505-11.
doi: 10.1194/jlr.D034553. Epub 2013 Feb 21.

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

Jessica Larose  1 , Pierre Julien, Jean-François Bilodeau
Affiliations

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

Jessica Larose et al. J Lipid Res. 2013 May.

Abstract

Plasma F2-isoprostanes (F2-isoPs) are reliable biomarkers of oxidative stress. Several possible F2-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 F2-isoPs, namely 8-iso-15(R)-prostaglandin F2α (PGF2α), 8-iso-PGF2α, 15(R)-PGF2α, iPF2α-IV, iPF2α-VI, 5-iPF2α-VI, and (±)5-8,12-iso-iPF2α-VI. We have validated this method in plasma of pregnant women, a mild physiological oxidative stress known to increase F2-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 F2-isoPs. The F2-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 F2-isoPs in plasma of pregnant women (P < 0.05). The 15(R)-PGF2α 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 F2-isoP regioisomers.

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Figures

Fig. 1.
Fig. 1.
Chemical structures of analytes investigated by LC-MS/MS.
Fig. 2.
Fig. 2.
Mass chromatograms of a standard solution (A–F) and of a typical plasma sample spiked with 50 pg of each internal standard (G–L). Peak identification (see Fig. 1 for structure): 1, 8-iso-15(R)-PGF2α; 2, 8-iso-PGF2α; 3, 8-iso-PGF2β; 4, 11β-PGF2α; 5, 15(R)-PGF2α; 6, 5-trans-PGF2α; 7, PGF2α; 8, 8-iso-PGF2α-d4; 9, PGF2α-d4; 10, iPF2α-IV; 11, iPF2α-IV-d4; 12, iPF2α-VI; 13, 5-iPF2α-VI; 14, (±)5-8,12-iso-iPF2α-VI; 15, iPF2α-VI-d11; 16, 5-iPF2α-VI-d11; 17, (±)5-8,12-iso-iPF2α-VI-d11; ?, unknown compounds. IS, deuterated internal standards.
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