A Single Reversed-Phase UPLC Method for Quantification of Levofloxacin in Aqueous Humour and Pharmaceutical Dosage Forms

Abstract

An attempt was made to develop a single, rapid, specific, and sensitive gradient reversed-phase ultra-performance liquid chromatographic method for quantitative analysis of levofloxacin. The single method thus developed is applied for the quantification of levofloxacin both in aqueous humour as well as pharmaceutical dosage forms (i.e., tablets and eye drops). The newly developed method is applicable for pharmacokinetic studies of eye formulations. The chromatographic separation of levofloxacin was achieved on a Waters Acquity HSS T-3 column (100 x 2.1 mm, 1.8 microm) within a short run-time of 5 min. The method was validated according to the ICH guidelines with respect to system suitability, linearity, limit of quantitation and detection, precision, accuracy, robustness, and specificity. Forced degradation studies were also performed in levofloxacin bulk drug samples to demonstrate the stability-indicating power of the developed ultra-performance liquid chromatography method. The developed method was then successfully applied for the ocular pharmacokinetic study of levofloxacin eye formulations and assay of levofloxacin pharmaceutical dosage form.