title: Solid-phase extraction and high-performance liquid chromatography analysis of lipoxygenase pathway products.
abstract: Liquid chromatography methods for quantitation of leukotrienes and HETEs (hydroxyeicosatetraenoic acids) in biological samples are described. Extraction is accomplished by acetonitrile precipitation of proteins followed by selective acetonitrile elution from a solid-phase C18 extraction cartridge. Isocratic elution of extracts from short reverse-phase columns with 3 microns C18-bonded silica results in elution of all components of interest in less than 10 min. The addition of the mobile-phase additives, trifluoroacetic acid and triethylamine, enable the peptido-leukotrienes to be eluted with excellent peak shape and unique elution times. As little as 1 pmol of each metabolite can be detected by uv spectrophotometry with a wavelength change from 280 to 235 nm midway through the chromatographic run. This method is demonstrated by the extraction and analysis of leukotriene and HETE standards from human serum.
mesh_terms: D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D001091:Arachidonate Lipoxygenases; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005021:Ethylamines; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis* / Q000097:blood; D007975:Leukotriene B4 / Q000032:analysis* / Q000097:blood; D008084:Lipoxygenase; D013189:SRS-A / Q000032:analysis* / Q000097:blood; D014269:Trifluoroacetic Acid
chemical_list: D005021:Ethylamines; D006893:Hydroxyeicosatetraenoic Acids; D013189:SRS-A; 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; D014269:Trifluoroacetic Acid; D001091:Arachidonate Lipoxygenases; D008084:Lipoxygenase; C016162:triethylamine