title: Platelet-Activating Factor Quantification Using Reversed Phase Liquid Chromatography and Selected Reaction Monitoring in Negative Ion Mode.
abstract: Platelet-activating factor (PAF) is a potent biologically active phospholipid that mediates human physiological and pathophysiologic responses. PAF levels increase transiently and are typically assessed by techniques with limitations related to expense, sensitivity, pre-analysis derivatization and interference with isobaric molecules. This study elucidates a facile, accurate liquid chromatography-mass spectrometry analytical method for PAF. In negative ion mode using electrospray ionization, collisionally-activated dissociation analysis showed a unique product ion for acetate adducts of PAF molecular species representing the loss of methyl acetate from the polar head group and loss of a part of the acetate group from the sn-2 position. This product ion was exploited for selected reaction monitoring of PAF molecular species following separation by reversed-phase liquid chromatography. Standard calibration responses were determined, and this method was able to detect as low as 100 fmol of PAF. Finally, PAF molecular species were quantified in human neutrophils and monocytes.
authors.affiliation: Edward A. Doisy Department of Biochemistry and Molecular Biology, and Center for Cardiovascular Research, Saint Louis University School of Medicine, St. Louis, MO, 63104, USA.
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Pike
authors.affiliation: Edward A. Doisy Department of Biochemistry and Molecular Biology, and Center for Cardiovascular Research, Saint Louis University School of Medicine, St. Louis, MO, 63104, USA.
authors.forename: Celine L
authors.initials: CL
authors.lastname: Hartman
authors.affiliation: Edward A. Doisy Department of Biochemistry and Molecular Biology, and Center for Cardiovascular Research, Saint Louis University School of Medicine, St. Louis, MO, 63104, USA.
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Weissler
authors.affiliation: Edward A. Doisy Department of Biochemistry and Molecular Biology, and Center for Cardiovascular Research, Saint Louis University School of Medicine, St. Louis, MO, 63104, USA.
authors.forename: Elisa N D
authors.initials: EN
authors.lastname: Palladino
authors.affiliation: Edward A. Doisy Department of Biochemistry and Molecular Biology, and Center for Cardiovascular Research, Saint Louis University School of Medicine, St. Louis, MO, 63104, USA.
authors.forename: Carolyn J
authors.initials: CJ
authors.lastname: Albert
authors.affiliation: Edward A. Doisy Department of Biochemistry and Molecular Biology, and Center for Cardiovascular Research, Saint Louis University School of Medicine, St. Louis, MO, 63104, USA. fordda@slu.edu.
authors.forename: David A
authors.initials: DA
authors.lastname: Ford
mesh_terms: D000085:Acetates / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D006801:Humans; D009000:Monocytes / Q000378:metabolism*; D009504:Neutrophils / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000032:analysis* / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000085:Acetates; D010972:Platelet Activating Factor
keywords: Mass spectrometry; Monocytes; Neutrophils; Platelet-activating factor