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A more recent version of this article appeared on February 1, 2005

Papers In Press, published online ahead of print November 1, 2004
J. Lipid Res., doi:10.1194/jlr.D400029-JLR200
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Submitted on October 13, 2004
Revised on November 1, 2004
Accepted on October 28, 2004

An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry

John S. Owen, Robert L. Wykle, Michael P. Samuel, and Michael J. Thomas

Biochemistry Dept., Wake Forest University School of Medicine, Winston-Salem, NC 27157

Corresponding Author: joowen{at}wfubmc.edu

We describe an improved assay for platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) using HPLC-tandem mass spectrometry (LC-MS/MS). The present method can readily detect as little as 1 pg (1.9 fmol) of PAF, a significant improvement over previously described LC-MS/MS methods, and gives a linear response up to 1000 pg of PAF. Our method also overcomes the artifacts from isobaric lipids which have limited the usefulness of certain existing LC-MS/MS assays for PAF. In the course of these studies, we detected three novel lipid species in human neutrophils. One of the novel lipids appears to be a new molecular species of PAF, and the other two have chromatographic and mass spectrometric properties consistent with stearoyl-formyl-glycerophosphocholine and oleoyl-formyl-glycerophosphocholine. These observations identify previously unknown potential interferences in the measurement of PAF by LC-MS/MS. Moreover, our data suggest that the previously described palmitoyl-formyl-glycerophosphocholine is not unique, but rather is a member of a new and poorly understood family of formylated lipids.


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