High-performance liquid chromatographic analysis of three macrolide antibiotics in pharmaceutical formulations and in human plasma

ABSTRACT

A high-performance liquid chromatographic method, for the determination of three macrolide antibiotics [roxithromycin [RXE], erythromycin base [E] and erythromycin ethylsuccinate [EES]], is presented. Study of the optimum condition for separation and quantitation of these antibiotics indicated that chromatography is performed in the reversed-phase mode using a C-18 column at a temperature of 40C. The strength of phosphate buffer can be used to control selectivity of the separated compound. The addition of isopropanol to the mobile phase improves peak shapes for the compounds of interest. Validation data for the studied antibiotics is done on spiked human plasma samples after extraction by adopting a rapid and simple procedure. The intra- and inter-assay coefficients of variation were less than 5% for the spiked plasma and pharmaceutical dosage forms