Analysis of platelet-activating factor by GC-MS after direct derivatization with pentafluorobenzoyl chloride and...

Parallel analysis of platelet-activating factor (PAF) using chemical ionization gas chromatography-mass spectrometry after direct derivatization with pentafluorobenzoyl chloride (PFB) and heptafluorobutyric anhydride (HFB) provides a facile and highly sensitive means for detecting and elucidating the structure of the numerous alkyl-chain homologs of this acetylated phospholipid autacoid. In the present study, the PFB derivative was used for initial electron capture negative ion chemical ionization analysis of PAF candidate molecules in human PMN extracts of unknown composition. Subsequent pulsed positive ion/electron capture negative ion chemical ionization evaluation of the HFB derivative furnished a measure of the molecular weight from [MH]+ and yielded the required structural information from characteristic negative ions, in particular [M-(2HF + ketene)]- and [M-(HF + acetic acid)]-. These procedures easily permitted confirmation of the presence of C16:0-, C17:0-, C18:0-, and C18:1-AGEPC (acetyl glyceryl ether phosphocholine) in extracts of stimulated human PMN and also demonstrated that the C17:0- homolog was comprised of both straight-chain and branch-chain varieties.