Quantitation of lyso-platelet activating factor molecular species from human neutrophils by mass spectrometry.

摘要: Two physicochemical methods have been developed for the quantitative analysis of lyso-platelet activating factor (lyso-PAF) based on gas-liquid chromatography-mass spectrometry (GLC/MS) and fast atom bombardment-mass (FAB/MS) using stable isotope dilution. After addition deuterated internal standards, lyso-PAF produced from neutrophils was purified by silicic acid chromatography thin-layer (TLC). The GLC/MS assay employed phospholipase C or hydrofluoric hydrolysis phosphocholine moiety to yield ether monoglycerides. Condensation monoglycerides with acetone yielded 1-O-alkyl-2,3-isopropylidene glycerol which could be analyzed GLC/MS. ions corresponding M-15 fragments both labeled unlabeled derivatives were monitored in a selected ion recording mode. Standard curves found linear over range tested (10-2000 ng) limit detection below 200 pg injected column. For FAB/MS assay, unmodified well suited direct analysis; however, (S/N greater than 3) matrix 5 ng placed probe tip. It that human contain approximately 300 pg/10(6) cells increased 2-3-fold during 5-min period following challenge 1.9 microM calcium ionophore, A23187. molecular species identified as hexadecyl octadecyl ethers at sn-1 predominating abundance after stimulation.