Isocratic determination of arachidonic acid 5-lipoxygenase products in human neutrophils by high-performance liquid chromatography.
A high-performance liquid chromatographic method was developed to determine arachidonic 5-lipoxygenase products in calcium ionophore-stimulated neutrophils. This procedure allows the simultaneous measurement of leukotriene B4 (LTB4) and its omega-oxidation products without using a gradient elution system. 20-Carboxy-LTB4, 20-hydroxy-LTB4, 6-trans-LTB4, 12-epi-6-trans-LTB4, LTB4 and 5s,12s-dihydroxyeicosatetraenoic acid can be separated and quantitated by reversed-phase chromatography using isocratic elution. The generation and degradation of 5-lipoxygenase products by human neutrophils following stimulation with calcium ionophore have been examined by this method.
Mita
H
H
Clinical Research Centre for Rheumato-Allergology, National Sagamihara Hospital, Kanagawa, Japan.
Yui
Y
Y
Yasueda
H
H
Shida
T
T
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D001094:Arachidonate 5-Lipoxygenase / Q000032:analysis*; D001091:Arachidonate Lipoxygenases / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis; D007202:Indicators and Reagents; D007536:Isomerism; D007975:Leukotriene B4 / Q000032:analysis; D009504:Neutrophils / Q000201:enzymology*; D010084:Oxidation-Reduction; D013056:Spectrophotometry, Ultraviolet
D006893:Hydroxyeicosatetraenoic Acids; D007202:Indicators and Reagents; D007975:Leukotriene B4; C022022:5-hydroxy-6,8,11,14-eicosatetraenoic acid; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; C025984:15-hydroxy-5,8,11,13-eicosatetraenoic acid; D001091:Arachidonate Lipoxygenases; D001094:Arachidonate 5-Lipoxygenase