Determination of platelet-activating factor by a chemiluminescence method and its application to stimulated guinea pig neutrophils.

A simple, rapid and sensitive chemiluminescence method has been developed to measure platelet-activating factor (PAF). Hydrogen peroxide generated from PAF, upon phospholipase D cleavage, by choline oxidase is determined as chemiluminescence by a luminol-microperoxidase system. The detection limit of PAF by this method is 5 pmol/tube. The method is reproducible with a 5.5% coefficient of variation at 10 pmol of PAF (n = 5). Lipids were extracted from guinea pig neutrophils after stimulation with cytochalasin B and N-formyl-methionyl-leucyl-phenylalanine, and PAF was isolated by high-performance liquid chromatography and determined by chemiluminescence measurements. The amount of PAF detected was 96.1 +/- 39.7 (mean +/- SD, n = 7) pmol/10(8) cells. This highly sensitive method could be useful for the determination of PAF generated under pathophysiological conditions.