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Updated: May 10, 2026
Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis
Published on: November 17, 2011
HPLC method for determination of lipoxygenase positional specific products
Peter Hoffman1, Drahomíra Rauová, Lýdia Bezáková
1Faculty of Pharmacy, Department of Cellular and Molecular Biology of Drugs, Comenius University in Bratislava, Bratislava, Slovak Republic. hoffman@fpharm.uniba.sk
Abstract:
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.
