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

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