Korean J Physiol Pharmacol.  1998 Dec;2(6):779-786.

Simultaneous HPLC analysis of arachidonic acid metabolites in biological samples with simple solid phase extraction

• Kim HG
• Huh YN
• Park KS

Abstract

A reversed-phase high-performance liquid chromatography (RP-HPLC) has been developed to analyze the metabolites of arachidonic acid based on the specificities of ultraviolet absorption of these various metabolites and is sensitive to the nanogram level. This procedure makes it possible to extract complex mixtures of eicosanoids efficiently with a single step and to analyze them simultaneously by RP-HPLC from biological samples using octadesylsilyl silica extraction column and PGB2 as an internal standard. The cyclooxygenase, products (prostaglandin (PG)D2, PGE1, PGE2, PGF1alpha, PGF2alpha, 6-keto-PGF1alpha, and thromboxane B2 (TXB2)) and lipid peroxidation product, isoprostanes, of arachidonic acid were monitored by one isocratic HPLC system at 195 nm wavelength. The lipoxygenase products (leukotriene(LT)B4, LTC4, LTD4, and 5-hydroxyeicosatetraenoic acid (5-HETE), 12-HETE, 15-HETE) were measured by another isocratic HPLC system at 280 nm for LTs and 235 nm for HETEs. This method provides a simple and reliable way to extract and assess quantitatively the final arachidonic acid metabolites.

Keyword

High-performance liquid chromatography; Arachidonic acid; Eicosanoid; Prostaglandin; Leukotriene; Hydroxyeicosatetraenoic acid; Isoprostane; Solid phase extraction

MeSH Terms

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid
Absorption
Alprostadil
Arachidonic Acid*
Chromatography, High Pressure Liquid*
Chromatography, Liquid
Complex Mixtures
Dinoprost
Dinoprostone
Eicosanoids
Hydroxyeicosatetraenoic Acids
Isoprostanes
Leukotriene C4
Leukotriene D4
Lipid Peroxidation
Lipoxygenase
Prostaglandin-Endoperoxide Synthases
Silicon Dioxide
Solid Phase Extraction*
Thromboxane B2
12-Hydroxy-5,8,10,14-eicosatetraenoic Acid
Alprostadil
Arachidonic Acid
Complex Mixtures
Dinoprost
Dinoprostone
Eicosanoids
Hydroxyeicosatetraenoic Acids
Isoprostanes
Leukotriene C4
Leukotriene D4
Lipoxygenase
Prostaglandin-Endoperoxide Synthases
Silicon Dioxide
Thromboxane B2