Abstract
A solid-phase extraction technique for the isolation of platelet activating factor [1-O-alkyl-2-acetyl- sn-glycero-3-phosphocholine (AGEPC)] from biological matrices was developed. Amberlite XAD-2 was effective in retaining different molecular species of AGEPC from plasma and incubation media. The recovery of the three molcular species (C 14-, C 16-, and C 18-AGEPC) was greater than 95%. XAD-2 also removed large amounts of plasma impurities, giving a cleaner high-performance liquid chromatographic (HPLC) profile. AGEPC in plasma or incubation media was not significantly removed by passage of the sample through a column packed with ODS-silica. A reversed-phase HPLC technique for separation and estimation of different species of AGEPC was developed. Resolution of C 14-, C 16- and C 18-AGEPC was accomplished on a Hamilton PRP-1 resin column using an aqueous acetonitrile gradient containing 1 m M methanesulphonic acid. The detection limit was of the order of 50 ng of AGEPC at 210 nm. The AGEPC purified by the technique described retained its biological activity as determined by its ability to release endogenous serotonin from rabbit platelets.
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