Mass spectrometric analysis of platelet-activating factor after isolation by solid-phase extraction and direct derivatization with pentafluorobenzoic anhydride.

S T Weintraub; R K Satsangi; E A Sprague; T J Prihoda; R N Pinckard

doi:10.1016/s1044-0305(99)00135-x

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Mass spectrometric analysis of platelet-activating factor after isolation by solid-phase extraction and direct derivatization with pentafluorobenzoic anhydride.

S T Weintraub·R K Satsangi·E A Sprague·T J Prihoda·R N Pinckard

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Abstract

Platelet-activating factor is the term used to denote a class of extremely potent lipid mediators that consist predominantly of 1-O-alkyl- and 1-O-acyl-2-acetyl-sn-glycero-3-phosphocholines. A method has been devised for rapid isolation of these acetylated phospholipids by solid-phase extraction prior to direct derivatization with pentafluorobenzoic anhydride and analysis by gas chromatography (GC)/electron-capture mass spectrometry. Recovery through the entire method (lipid isolation, derivatization, and purification) typically ranged from 70% to 85%. Using the direct derivatization procedure described here, the practical limit of detection for each of the standard alkyl- and acyl-platelet-activating factor homologs was 1 fmol injected into the GC. Results from the application of the method to the analysis of alkyl and acyl homologs of platelet-activating factor isolated from stimulated human umbilical vein endothelial cells are presented, exhibiting excellent accuracy and precision for a wide range of tissue levels of this class of potent autacoids.

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Anhydrides Benzoates Cells, Cultured Endothelium, Vascular Gas Chromatography-Mass Spectrometry Humans Indicators and Reagents Mass Spectrometry Platelet Activating Factor Fluorobenzenes Fluorocarbons Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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S T Weintraub, R K Satsangi, E A Sprague, T J Prihoda, R N Pinckard. 2000. Mass spectrometric analysis of platelet-activating factor after isolation by solid-phase extraction and direct derivatization with pentafluorobenzoic anhydride.. https://doi.org/10.1016/s1044-0305(99)00135-x

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