Detection of lyso-platelet-activating factor by high-performance liquid chromatography after derivatisation with fluorescent fatty acids.
Lyso-platelet-activating factor (lyso-PAF) was derivatised with 9-anthracenepropionic acid in the presence of dicyclohexylcarbodiimide, p-toluenesulfonic acid and 4-dimethylaminopyridine. The reaction yield exceeded 90% when the fatty acid was present in double molar amounts versus lyso-PAF. The procedure was equally effective in the derivatisation of other lysophospholipids. The derivatized phospholipids are detected by ultraviolet absorption (lambda = 253 nm) or fluorescence detection (using excitation at 254 nm and emission at 450 nm). The technique was applied successfully to the detection of lyso-PAF in complement activated rabbit plasma.
Salari
H
H
Department of Medicine, University of British Columbia, Vancouver, Canada.
Eigendorf
G K
GK
D015761:4-Aminopyridine / Q000031:analogs & derivatives; D000818:Animals; D000873:Anthracenes; D001557:Benzenesulfonates; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004024:Dicyclohexylcarbodiimide; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D010972:Platelet Activating Factor / Q000031:analogs & derivatives* / Q000032:analysis; D011817:Rabbits
D000873:Anthracenes; D001557:Benzenesulfonates; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; C029271:O-deacetyl platelet activating factor; D010972:Platelet Activating Factor; C065158:9-anthracenepropionic acid; D004024:Dicyclohexylcarbodiimide; D015761:4-Aminopyridine; C003885:4-dimethylaminopyridine; C029501:4-toluenesulfonic acid