Faculty Collaboration Database

An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry. J Lipid Res 2005 Feb;46(2):373-82

Date

11/03/2004

Pubmed ID

15520455

DOI

10.1194/jlr.D400029-JLR200

Scopus ID

2-s2.0-13944253190 (requires institutional sign-in at Scopus site)   20 Citations

Abstract

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 1,000 pg of PAF. Our method also overcomes the artifacts from isobaric lipids that 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.

Author List

Owen JS, Wykle RL, Samuel MP, Thomas MJ

Author

Michael J. Thomas PhD Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Catalysis
Chromatography
Chromatography, High Pressure Liquid
Clinical Chemistry Tests
Glycerophospholipids
Humans
Lipid Metabolism
Lipids
Lysophosphatidylcholines
Mass Spectrometry
Neutrophils
Platelet Activating Factor
Sensitivity and Specificity
Time Factors