HPLC method for determination of lipoxygenase positional specific products.

Mammalian lipoxygenases (LOXs) play an important role in physiological and pathological processes through the biosynthesis of lipid mediators-leukotrienes, lipoxins and other arachidonic acid derivatives.There are four major families of LOXs that can be analyzed through the production of hydroxyeicosatetraenoic acids (HETEs). No analytical method to detect 5-, 8-, 12- and 15-HETE in one run has been published to date. The HPLC method combines reversed-phase separative column Nucleosil 120-5 C18 and NP column Zorbax Rx.SIL for identification. This conjunction enables separation of 12-HETE and 15-HETE to the baseline which is essential in 12/15-LOX research and elution of all four HETEs in one run. The method was successfully tested on partially purified LOXs from rat lung cytosol.