Analysis of human platelet arachidonate metabolism by high performance liquid chromatography （HPLC）

In order to evaluate whether the arachidonic acid metabolism of the platelets from patients with platelet dysfunction is nomal or not, the analysis of the metabolites via cyclo-oxygenase and lipoxygenase pathway was done, using high performance liquid chromatography（HPLC）. Reversed phase HPLC for analysis of the lipoxygenase products revealed 5－HETE, 12－HETE and 15－HETE in the supernatants from normal washed human platelets induced by arachidonic acid（0.2mM）. PGD_2, TXB_2 and PGE_2, which are the cyclo-oxygenase products, were also identified in the supernatants using 9－anthryl-diazomethane derivatization of prostaglandins for normal phase HPLC. Although 5－HETE and 15－HETE were produced by the contaminated polymorphonuclear leukocytes, the metabolites of arachidonic acid could be separated well and be detected at high sensitivity. According to these facts, the assay method using HPLC could be a useful tool for not only the detection of a series of arachidonic acid metabolites in human platelets, but also the diagnosis of the patients who show platelet function abnormality