High-Performance Liquid Chromatography for the Simultaneous Estimation of Cefoperazone and Sulbactam in Rat Plasma and its Importance in Therapeutic Drug Monitoring

Abstract

Therapeutic drug monitoring and pharmacokinetic studies of antibiotics need a sensitive and precise measurement ofplasma drug concentrations. In the present study, we developed a rapid, sensitive and selective chromatographicmethod for simultaneous estimation of cefoperazone (CEF) and sulbactam (SAL) in male Wistar rat plasma. A novelliquid phase extraction method has adopted the preparation of plasma sample preparation. The CEF and SAL wereeluted on a peerless Basic C18 (25 cm; 4.6 mm x 5 μm) column maintained at controlled environmental conditions.The gradient mobile phase comprised of 10 mM ammonium acetate and acetonitrile. A UV detector was set at 250nm and retention times for CEF and SAL were approximately 5.6 and 14.2 min, respectively. The proposed HPLCmethod was validated according to the US FDA guidelines with respect to the linearity, accuracy, precision, detectionand quantitation limits, robustness and specificity. Calibration curves of CEF and SAL were linear across theconcentration range of 600-1000 and 6-10 μg/mL, with correlation coefficients (r2) >0.9977 and (r2) >0.9987,respectively. The limits of detection for CEF and SAL were 70.48 and 0.35 μg/mL, respectively. Additionally, CEFand SAL were stable in plasma for at least 24 h when stored at room temperature and 2-8 C. Furthermore, thedeveloped chromatographic method was effectively utilized to measure the plasma CEF and SAL concentrations in apharmacokinetics study after intravenous injection to the healthy male Wistar rats.