Quantification of Leukotriene B4 Using High-Performance Liquid Chromatography

Abstract

Leukotriene B₄ (LTB₄) was derivatized with p-(9-anthroyloxy) phenyl bromide forming the panacyl ester. The yield from this reaction was 95 percent. Subsequent separation of this ester using reversed phase high-performance liquid chromatography resulted in a greater than 10 fold increase in sensitivity as measured directly with ultraviolet light (absorption at 254 nm). Sensitivity was 16 ng and elution time was less than 15 minutes. We applied this technique to quantification of LTB₄ production by human polymorphonuclear leucocytes.