ISSN:
0887-6134
Keywords:
Chemistry
;
Analytical Chemistry and Spectroscopy
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Chemistry and Pharmacology
Notes:
Platelet activating factor (PAF) bioactivity has been demonstrated in saliva from normal volunteers. We sought structural confirmation and evidence of heterogeneity in the 1-O-alkyl chain of the acetyl glyceryl ether phosphoryl choline (AGEPC) extracted from saliva by employing stable isotope dilution techniques in conjunction with gas chromatography/mass spectrometry. The method described involves removal of the polar phosphocholine moiety, accounts for acetyl group migration, and allows for acylation of the resultant free hydroxyl with pentafluorobenzoyl chloride. Thin-layer chromatography (TLC) purification is undertaken after phospholipase C cleavage and again after pentafluorobenzoyl chloride derivatization. The majority of the ion current is represented in the molecular anion, allowing measurement of 50 pg in biological fluid with a signal-to-noise ratio of ≥9. In one subject with markedly increased salivary PAF levels, we found evidence for molecular heterogeneity of AGEPC with production of not only C16:0 but also C18:0 and C18:1 in the alkyl chain. This technique, by using TLC in lieu of high-performance liquid chromatography, avoids potentially confounding trace contamination effects, produces spectra with few interfering signals and increases sample throughput.
Additional Material:
5 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/bms.1200180409
