Journal of Lipid Research, Vol 25, 753-757, Copyright © 1984 by Lipid Research, Inc.
High performance liquid chromatography of platelet-activating factors
EM Jackson, GE Mott, C Hoppens, LM McManus, ST Weintraub, JC Ludwig and RN Pinckard
Silica and C18 reverse phase high performance liquid chromatography (HPLC)
were used to fractionate synthetic molecular species of 1-O-
alkyl-2-acetyl-sn-glycero-3-phosphocholine (AGEPC) and semi-synthetic
platelet-activating factor (PAF) synthesized from beef heart plasmalogens.
A single coincident peak from silica HPLC was observed for either a mixture
of synthetic AGEPC's with alkyl chain lengths from C12 to C18 or for beef
heart-derived PAF. This peak was well separated from other classes of
phospholipid standards including 2- lysophosphatidylcholine and 3H-labeled
lyso-PAF. Subsequently, the synthetic AGEPC mixture or beef heart PAF was
separated into individual species on a C18 reverse phase column. Beef
heart-derived PAF was fractionated into at least four molecular species of
PAF activity which had similar retention times as the radioactivity of
3H-labeled beef heart PAF. Approximately 56% of the radioactivity of
3H-labeled PAF was found in the fraction with a similar retention time as
1-O-hexadecyl-2- acetyl-sn-glycero-3-phosphocholine, 10% as
1-O-octadecyl-2-acetyl-sn- glycero-3-phosphocholine, 11% as
1-O-pentadecyl-2-acetyl-sn-glycero-3- phosphocholine, and 13% in an
unidentified fraction which eluted after C-16-AGEPC. The unidentified
fraction did not correspond to any of the homologous series of synthetic
AGEPCs with saturated alkyl chain lengths from C12 to C18. Recoveries of
radioactive phospholipids from silica or reverse phase columns were greater
than 95%.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
