Chromatographic Analyses of Ether-Linked Lipids Involved in PAF Metabolism
A number of chromatographic separations were essential in elucidating the structure of l-alkyl-2-acetyl-sn-glycero-3-phosphocholine [(alkylacetyl-GPC) platelet activating factor (PAF)] (Benveniste et al., 1979; Blank et al., 1979; Demopoulos et al., 1979; Hanahan et al., 1980) and chromatography remains an indispensable tool for all biochemical studies involving PAF. The three main types of chromatography used in PAF research are (1) open-column chromatography with columns packed with silicic acid, (2) thin-layer chromatography (TLC) on layers of silica gel, and (3) high-performance liquid chromatography (HPLC) with both normal- and reverse-phase columns. Rather than trying to describe the myriad of modifications of solvent systems that have appeared in the literature, we attempt to review what we consider as basic chromatographic systems for the analyses and isolation of PAF. Because it is important to determine the particular species of alkylacyl-GPC as precursors (reactions catalyzed by phospholipase A2 and acetyl-CoA: acetyltrans-ferase) in the biosynthesis of PAF and the molecular origin of the acyl moieties that acylate alkyllyso-GPC (the latter formed by acetylhydrolase inactivation of PAF), we describe a number of practical chromatographic methods that can be used to identify the subclasses and specific molecular species of the diradyl phospholipids involved. Chromatographic methods that we have used for analysis of alkylacetyl-glycerols (an alternate precursor of PAF) and their metabolites are also described in this chapter.
KeywordsMolecular Species Glyceryl Ether Rabbit Platelet Individual Molecular Species Phospholipid Molecular Species
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