Developing and Validating a Stability-Indicating Method for the Analysis of Piperacillin and Tazobactam in Bulk and Dosage Forms in Human Plasma Using RP-HPLC.

Abstract

A meticulously developed and thoroughly validated High-Performance Liquid Chromatography (HPLC) method was designed for the simultaneous determination of Piperacillin and Tazobactam in a Fixed-Dose Combination. The chromatographic profiles were constructed using a mobile phase of Methanol: 0.1% orthophosphoric Acid (OPA) in water (85:15) with a flow rate of 0.7 ml/min. Employing a C18 Column (4.6 x 250 mm, 5μm particle size) as the stationary phase, detection at 231 nm facilitated reliable quantification. The method exhibited linear correlation within concentration ranges of 16-80 µg/ml for Piperacillin and 02-10 µg/ml for Tazobactam, with regression values of 0.9992 and 0.9999, respectively. Precision studies demonstrated % RSD values below 2% for both drugs across all selected concentrations. The limit of detection (LOD) and limit of quantification (LOQ) were determined as 0.258 μg/ml and 0.784 μg/ml for Piperacillin, and 0.143 μg/ml and 0.435 μg/ml for Tazobactam, respectively. Beyond method validation, stability studies subjected analyte solutions to acid, base, oxidative, and hydrolytic stress conditions. The validation process adhered to the guidelines outlined by the International Conference on Harmonization (ICH).