Mass spectrometric analysis of platelet-activating factor after isolation by solid-phase extraction and direct derivatization with pentafluorobenzoic anhydride.
Platelet-activating factor is the term used to denote a class of extremely potent lipid mediators that consist predominantly of 1-O-alkyl- and 1-O-acyl-2-acetyl-sn-glycero-3-phosphocholines. A method has been devised for rapid isolation of these acetylated phospholipids by solid-phase extraction prior to direct derivatization with pentafluorobenzoic anhydride and analysis by gas chromatography (GC)/electron-capture mass spectrometry. Recovery through the entire method (lipid isolation, derivatization, and purification) typically ranged from 70% to 85%. Using the direct derivatization procedure described here, the practical limit of detection for each of the standard alkyl- and acyl-platelet-activating factor homologs was 1 fmol injected into the GC. Results from the application of the method to the analysis of alkyl and acyl homologs of platelet-activating factor isolated from stimulated human umbilical vein endothelial cells are presented, exhibiting excellent accuracy and precision for a wide range of tissue levels of this class of potent autacoids.
Weintraub
S T
ST
Department of Biochemistry, University of Texas Health Science Center, San Antonio 78229-3900, USA. weintraub@uthscsa.edu
Satsangi
R K
RK
Sprague
E A
EA
Prihoda
T J
TJ
Pinckard
R N
RN
D000812:Anhydrides / Q000737:chemistry; D001565:Benzoates / Q000737:chemistry; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D010972:Platelet Activating Factor / Q000737:chemistry*; D005464:Fluorobenzenes; D005466:Fluorocarbons
D000812:Anhydrides; D001565:Benzoates; D007202:Indicators and Reagents; D010972:Platelet Activating Factor; C060431:pentafluorobenzoic anhydride; D005464:Fluorobenzenes; D005466:Fluorocarbons