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

• Published in: Journal of the American Society for Mass Spectrometry 2000-02, Vol.11 (2), p.176-181
• Main Authors: Weintraub, Susan T, Satsangi, Rajiv K, Sprague, Eugene A, Prihoda, Thomas J, Pinckard, R.Neal
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Anhydrides - chemistry; Benzoates - chemistry; Biological and medical sciences; Cells, Cultured; Endothelial cells; Endothelium, Vascular - cytology; Endothelium, Vascular - drug effects; Endothelium, Vascular - metabolism; Extraction; Fundamental and applied biological sciences. Psychology; Gas chromatography; Gas Chromatography-Mass Spectrometry; Glycerolipids, phospholipids; Homology; Humans; Indicators and Reagents; Investigative techniques, diagnostic techniques (general aspects); Lipids; Mass Spectrometry; Medical sciences; Miscellaneous. Technology; Other biological molecules; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Phospholipids; Platelet Activating Factor - chemistry
• ISSN: 1044-0305; 1879-1123
• DOI: 10.1016/S1044-0305(99)00135-X

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.