Abstract

STABILITY INDICATING RP-HPLC METHOD FOR THE DETERMINATION OF PIPERACILLIN AND TAZOBACTAM AND THEIR RELATED SUBSTANCES IN BULK AND PHARMACEUTICAL FORMULATION

Khyathi Navle*, B. Lakshmi Prasanna, Hema Sree Dometti and Shankar Moodu

ABSTRACT

A simple, sensitive and precise high performance liquid chromatographic method was developed for the determination of related substances of Piperacillin and Tazobactam in bulk and pharmaceutical formulation. The Impurities were well separated on a Hypersil ODS column (250mm X 4.6mm, 5 μm) by the gradient program using Phosphate buffer (pH 6.0) and Acetonitrile at a flow rate of 1.0 mL /min with detection wavelength at 220 nm. The developed method separates Piperacillin and Tazobactam from its related substances by 45 minutes run time. The calibration curve shows linearity in the concentration range of 0.3-75.5 μg/ml for Piperacillin, 0.3-3.3 μg/ml for Tazobactam, 0.4-3.1 μg/ml for Aminosulfinic acid, 0.4-17.2 μg/ml for Ampicillin, 0.4-7.9 μg/ml for D-phenylglycyl EDP,0.4-15.8 μg/ml for Piperacillin sulfoxide with a correlation coefficient 0.9997, 0.9993, 0.9998, 0.9999, 0.9992, 0.9996 respectively. The method was validated for specificity, linearity, accuracy, precision, robustness and solution stability. Recovery was found to be in the range of 75-110% and RSD is <2%. The proposed method was successfully applied for the determination of Piperacillin and Tazobactam and their related substances in bulk and pharmaceutical formulations.

Keywords: Piperacillin(PIPER), Tazobactam(TAZO), HPLC, Related substances, Method Development.