DEVELOPMENT AND VALIDATION OF HPLC METHOD FOR THE ESTIMATION OF ANTIMICROBIAL DRUGS IN COMBINED DOSAGE FORM

Abstract

A simple, precise, accurate, and robust reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous estimation of Enmetazobactam (EZB) and Cefepime (CFP) in combined dosage form. Chromatographic separation was achieved on a C18 column (250 mm × 4.6 mm, 5 µm) using a mobile phase of methanol and acetonitrile (50:50 v/v) under isocratic conditions at a flow rate of 1.0 mL/min with detection at 254 nm. The retention times for EZB and CFP were found to be 3.75 min and 4.92 min, respectively. The method exhibited excellent linearity in the concentration range of 5–25 µg/mL for EZB and 10–50 µg/mL for CFP, with correlation coefficients (r²) of 0.9991 and 0.9990, respectively. The percentage recoveries ranged from 98.19% to 98.73%, confirming the accuracy of the method. Precision and robustness studies showed %RSD values less than 2%, indicating high method reproducibility. The LOD and LOQ values were 0.35 µg/mL and 0.85 µg/mL for EZB, and 0.50 µg/mL and 1.50 µg/mL for CFP, respectively. The developed method was successfully applied to the assay of marketed formulations, with assay values of 99.77% for EZB and 99.31% for CFP. Hence, the proposed RP-HPLC method is rapid, reliable, and suitable for routine quality control analysis of Enmetazobactam and Cefepime in pharmaceutical dosage forms.

KEYWORDS:

Enmetazobactam; Cefepime; RP-HPLC; Method Validation; Linearity; Precision; Recovery; LOD; LOQ; Combined Dosage Form.