Analysis of platelet-activating factor by gas chromatography-mass spectrometry: low-energy electron impact of the corresponding 3-acetyl-2-tert.-butyldimethylsilyl derivative.
A method for the analysis of platelet-activating factor in platelets employing gas chromatography and selected-ion monitoring mass spectrometry with low-energy electron impact and stable isotope dilution was developed. The procedure involved Bligh and Dyer extraction of the sample followed by thin-layer chromatographic purification. Platelet-activating factor is successively hydrolysed to the corresponding 2-acetyl-1-O-alkylglycerol by digestion with phospholipase C, and the product is allowed to isomerize to the more thermodynamically stable 3-acetyl-1-O-alkylglycerol before column purification and derivatization of the free OH with tert.-butyldimethylchlorosaline-imidazole. This reagent is of common use in platelet-activating factor derivatization, but is made to react with 2-acetyl instead of 3-acetyl isomer. The advantages of using the latter for the final derivatization are discussed and this method is compared with others currently available for gas chromatographic-mass spectrometric analysis of platelet-activating factor.
Triolo
A
A
Research Division Analytical Department, A. Menarini Industrie Farmaceutiche Riunite S.R.L., Florence, Italy.
Bertini
J
J
Mannucci
C
C
Perico
A
A
Pestellini
V
V
D001792:Blood Platelets / Q000737:chemistry; D004583:Electrons; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007477:Ions; D010972:Platelet Activating Factor / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000378:metabolism*
D007477:Ions; D010972:Platelet Activating Factor