Measurement of urinary leukotrienes by reversed-phase liquid chromatography and radioimmunoassay

Abstract

Leukotriene (LT) E4, an important LT metabolite appearing in urine, can be rapidly separated from normal and pathological urines by automated reversed-phase HPLC after a simple sample-processing. The recoveries of LTE4 afforded by this system (86.4 ± 6.5%, mean ± SEM for 60 ng/L, 85.4 ± 0.3% for 200 ng/L) are superior to those obtained by a manual extraction method. Consistency of results is similar. Highly reproducible retention times combined with a radioimmunoassay allow one to identify (based on co-elution) and quantify as little as 8 ng/L LTE4 in a 10-mL urine sample. LTE4 concentrations in urine from healthy persons approach this value (17 ± 5 ng/L), whereas samples from patients with cardiac ischemia show a wider range of concentrations (8 to 388 ng/L), up to 50 times the detection limit. Thus this method is applicable to the noninvasive investigation of leukotriene involvement in a wide range of ischemic, inflammatory, and hypersensitive conditions.