HPLC method for determination of lipoxygenase positional specific products

J Pharm Biomed Anal. 2013 Oct:84:53-8. doi: 10.1016/j.jpba.2013.05.041. Epub 2013 Jun 4.

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.

Keywords: Hydroxyeicosatetraenoic acid; Rat lung; Reversed-phase; Straight-phase; Tissue.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / chemistry*
  • Animals
  • Arachidonic Acid / chemistry*
  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, Reverse-Phase / methods
  • Hydroxyeicosatetraenoic Acids / chemistry*
  • Lipoxygenase / chemistry*
  • Lipoxygenase / metabolism*
  • Lung / enzymology
  • Lung / metabolism
  • Male
  • Rats
  • Rats, Wistar

Substances

  • Hydroxyeicosatetraenoic Acids
  • Arachidonic Acid
  • 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid
  • 15-hydroxy-5,8,11,13-eicosatetraenoic acid
  • Lipoxygenase