Abstract
METHOD DEVELOPMENT, VALIDATION AND SIMULTANEOUS ESTIMATION OF OFLOXACIN AND TINIDAZOLE APIS AND TABLET FORMULATION IN HUMAN PLASMA USING RP-HPLC

Ravichandran S.* and Gowtham Prabakaran

ABSTRACT

A superficial, selective, precise, and rapid isocratic reverse phase high performance liquid chromatography method with UV detection at 293nm was developed and validated for simultaneous estimation of Ofloxacin and Tinidazole as APIs and combined formulation from spiked human plasma. The column was equilibrated for at least 30min and separation was achieved by using Shimadzu-shim pack solar C8 (250×4.6mm, 5μm). The column was maintained at ambient temperature (25˚C). The chromatographic separation was performed with a mobile phase of acetonitrile: potassium dihydrogen orthophosphate buffer pH 4.5 (30:70v/v). The flow rate was 1mL/min, and the injector volume was 20μL. Ofloxacin and Tinidazole were well resolved from plasma constituents. The retention times was found to be 1.9 min for Ofloxacin and 3.2 min for Tinidazole. The method was found to be linear in the range of 1-5 μg/mL for Ofloxacin and 3-15 μg/mL for Tinidazole. The developed method was validated as per ICH guidelines. The developed precise method was validated in respect to precision, range, limit of detection, limit of quantitation, robustness as per ICH Q2B guidelines.

Keywords: Ofloxacin, Tinidazole, Plasma analysis, Bio-analytical method, RP-HPLC.


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