Monitoring eicosanoid biosynthesis via lipoxygenase and cyclooxygenase pathways in human whole blood by single HPLC run

Journal of Pharmaceutical and Biomedical Analysis
Petra FrohbergIvonne Wobst

Abstract

Eicosanoids play an important role as lipid mediators for physiological and pathological processes. Inhibitors of their biosynthesis have been developed as drugs for various diseases with major health political relevance. The search for more efficient inhibitors of eicosanoid formation requires simultaneous monitoring of various metabolic pathways. We developed an HPLC-based assay system, which quantifies lipoxygenase metabolites leukotriene B4 (LTB4), 5-hydroxyeicosatetraenoic acid (5-HETE), 12-hydroxyeicosatetraenoic acid (12-HETE), 15-hydroxyeicosatetraenoic acid (15-HETE) and cyclooxygenase metabolite 12-hydroxy-5,8,10-heptadecatrienoic acid (12-HHT) in whole human blood. Eicosanoid formation in blood is initiated with calcium ionophore A23187, arachidonic acid and calcium and magnesium ions. After solid phase extraction the different eicosanoids were separated by isocratic RP-HPLC using prostaglandin B1 as authentic standard. To verify the assay we determined the IC50 of known inhibitors of eicosanoid biosynthesis (zileuton, indomethacin, nordihydroguaiaretic acid). The test system is simple. It does not require extensive methodological experience and can be carried out in any biochemical laboratory. The analytical procedu...Continue Reading

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Citations

Feb 10, 2009·American Journal of Physiology. Lung Cellular and Molecular Physiology·Gautam DameraReynold A Panettieri
May 3, 2014·Journal of Pharmaceutical and Biomedical Analysis·Isabella SquellerioViviana Cavalca
Mar 12, 2010·Journal of Pharmaceutical and Biomedical Analysis·Ivana N BearaMira R Popović
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Apr 2, 2014·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Benedetta PorroViviana Cavalca
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May 1, 2020·Nutrients·Agnieszka StawarskaBarbara Bobrowska-Korczak
Jan 14, 2020·Journal of Trace Elements in Medicine and Biology : Organ of the Society for Minerals and Trace Elements (GMS)·Małgorzata JelińskaBarbara Bobrowska Korczak
Mar 13, 2014·Journal of Chemical Information and Modeling·Erchang ShangLuhua Lai
Apr 25, 2018·Journal of Medicinal Chemistry·Cong LiLuhua Lai

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