Abstract
Erythromycin is determined in both urine and plasma samples using microbore reversed-phase liquid chromatography with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)3(2+)] electrogenerated chemiluminescence (ECL) detection. Ru(bpy)3(2+) is included in the mobile phase thus eliminating band broadening caused by post-column reagent addition. Extra column band broadening is an important concern in microbore liquid chromatography due to the small peak volumes. Erythromycin was studied in both water and biological samples. The detection limit for erythromycin in standards is 0.01 microM or 50 fmol injected with a S/N of 3 and a linear working range that extends four orders of magnitude. Human urine and blood plasma were also studied. Urine samples were diluted and filtered before injection. Ultrafiltration was used to remove protein from blood plasma samples prior to injection. Erythromycin was selectively detected in the body fluid samples without any further sample preparation. The detection limits obtained for erythromycin in urine and plasma are 0.05 and 0.1 microM, respectively, for 5 microl injected on a 150x1 mm I.D. C18 column.
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