Separation of Hydroxyeicosatetraenoic Acids and Leukotrienes Originating from Kidney Tissue by C-18 Reverse-Phase High-Pressure Liquid Chromatography

Metabolism of arachidonic acid results in formation of eicosanoids including prostaglandins (PGs), thromboxanes (Txs), monohydroxyeicosatetraenocinoids (HETEs), leukotrienes (LTs), lipoxins (Lxs), epoxy metabolites, and hepoxilins (1 ). Most of these compounds are formed in minute quantities in response to specific stimuli and participate in modulation of the vascular tone or of the inflammatory process. We have been studying the formation of arachidonic acid metabolites in kidney tissue (mainly glomeruli) following immune injury. Normal glomeruli synthesize very small quantities of HETEs and LTs. Following immune injury, glomerular synthesis and levels of these eicosanoids increase (2 ). Leukotrienes (B₄ , C₄ , and D₄ ), 5-HETE, and 12-HETE can be separated, detected, and quantitated by several analytical techniques, including immunoassay, bioassay, gas chromatography-mass spectrometry (GC-MS), thin-layer chromatography (TLC), ultraviolet spectroscopy, and high pressure liquid chromatography (HPLC).