Separation and quantification of PGE 3 following derivatization with pancyl bromide by high pressure liquid chromatography with fluorometric detection

Abstract

Separation of prostaglandin E 3 (PGE 3) from prostaglandin E 2 (PGE 2) and prostaglandin E 1 (PGE 1) was achieved following derivatization with p-(9-anthroyloxy)phenacyl bromide (panacyl bromide). The eicosanoid esters were analysed by reverse phase high pressure liquid chromatography with fluometric detection (excitation 360nm and emission 470nm). Human, rat and mouse adherent cells were incubated overnight and the culture medium extracted, derivatized and analysed for PG production. PGE 2 was detected from biological samples of each species tested. PGE 2 synthesis was reduced when cells were incubated overnight with 5μM eicosapentaenoic acid. PGE 3 was not detectable under these experimental conditions. Studies were also undertaken using adherent cells from mice, rats and human subjects given dietary fish oil supplements rich in EPA. PGE 3 production by these cells was not detected although the dietary regimens yielded substantial incorporation of EPA into cell membranes and leukocyte LTB 5 production was demonstrable.