Simultaneous determination of lysophospholipids by high-performance liquid chromatography with fluorescence detection.
A high-performance liquid chromatography (HPLC) procedure for the separation of choline lysophospholipids including 1-acyl-lysophosphatidylcholines and 1-O-alkyl-lysophosphatidyl- cholines, like the lysoform of the platelet activating factor (2-lysoPAF), is described. The lysophospholipids are derivatized at the sn-2 position of the hydroxyl group by 7-diethylaminocoumarin-3-carbonylazide, which converts them into the corresponding carbamoyl derivatives. The derivatized compounds were well separated by reversed-phase HPLC and quantified by fluorimetric detection. This method shows a high sensitivity and allows the separation and quantification of mixtures of lysophospholipids at picomolar level. The method was applied to assay enzyme activities, like phospholipase A2 and PAF-acetylhydrolase, on single phospholipids or their mixtures.
Servillo
L
L
Dipartimento di Biochimica e Biofisica, II Università degli Studi di Napoli, Italy.
Iorio
E L
EL
Quagliuolo
L
L
Camussi
G
G
Balestrieri
C
C
Giovane
A
A
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005453:Fluorescence; D008246:Lysophospholipids / Q000032:analysis*; D000432:Methanol / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014867:Water / Q000737:chemistry
D008246:Lysophospholipids; D014867:Water; D000432:Methanol