Quantification of distinct molecular species of the 2-lyso metabolite of platelet-activating factor by gas chromatography-negative-ion chemical ionization mass spectrometry.
The biological activity of platelet-activating factor (PAF) is comprised by a few molecular species of phosphatidylcholine which contain a fatty alcohol connected by an ether linkage to the sn-1 position of the glycerol backbone and an acetate ester at the sn-2 position. The various molecular species of PAF differ in chain length and degree of unsaturation in the fatty alcohol residue side-chain. PAF is rapidly hydrolyzed to lyso-PAF by an acetylhydrolase enzyme which is quite active in a number of cells that synthesize PAF. We describe a method for quantitation of lyso-PAF which involves conversion to its propionate derivative in the presence of an internal standard (deuterium-labelled PAF), digestion to the diglyceride with Bacillus cereus phospholipase C, conversion to the pentafluorobenzoate derivative and capillary column gas chromatographic-negative-ion methane chemical ionization mass spectrometric analysis. Distinct molecular species of lyso-PAF can be individually quantitated at levels of 1 ng or less. These methods are applied to the demonstration of lyso-PAF accumulation in renal tissue from transplanted allografts undergoing acute rejection, in renal tissue from kidneys subjected to cold storage and autotransplantation, and in intestinal mucosa subjected to warm ischemia and reperfusion.
Turk
J
J
Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110.
Bohrer
A
A
Stump
W T
WT
Ramanadham
S
S
Mangino
M J
MJ
D000818:Animals; D003903:Deuterium; D004285:Dogs; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D007668:Kidney / Q000737:chemistry; D016030:Kidney Transplantation; D010972:Platelet Activating Factor / Q000031:analogs & derivatives* / Q000032:analysis; D014184:Transplantation, Homologous
C029271:O-deacetyl platelet activating factor; D010972:Platelet Activating Factor; D003903:Deuterium