Abstract
Objective: To study the Therapeutic drug monitoring and pharmacokinetics of marketed antibiotics formulation by developing a sensitive and specific Bioanalytical Chromatographic method.
 Methods: In the present study, we developed a rapid, sensitive and selective chromatographic method for simultaneous estimation of Cefoperazone (CEF) And Sulbactam (SAL) in male Wistar rat plasma. A novel liquid phase extraction method has adopted the preparation of plasma sample preparation. The CEF and SAL were eluted 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 mmol ammonium acetate and acetonitrile. A UV detector was set at 250 nm and retention times for CEF and SAL were approximately 5.6 and 14.2 min, respectively. The proposed HPLC method was validated according to the US FDA guidelines with respect to the linearity, accuracy, precision, detection and quantitation limits, robustness and specificity.
 Results: Calibration curves of CEF and SAL were linear across the concentration 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, CEF and SAL were stable in plasma for at least 24 h when stored at room temperature and 2-8 °C. 
 Conclusion: The developed chromatographic method was effectively utilized to measure the plasma CEF and SAL concentrations in a pharmacokinetics study after intravenous injection to the healthy male Wistar rats.
Highlights
Antibiotics are chemically synthesized antimicrobial molecules regularly prescribed over the times for the prevention and treatment of various infectious diseases
Appropriate dosing of the antibiotics is of utmost significance for clinical management
Attaining the correct therapeutic level is of high importance for avoiding the development of antimicrobial resistance
Summary
We developed a rapid, sensitive and selective chromatographic method for simultaneous estimation of Cefoperazone (CEF) And Sulbactam (SAL) in male Wistar rat plasma. The proposed HPLC method was validated according to the US FDA guidelines with respect to the linearity, accuracy, precision, detection and quantitation limits, robustness and specificity. CEF and SAL were as obtained as a gift sample from Emcure Bhosari, Pune. Cefoperazone-Sulbactam injection (1:0.5) purchased from a local medical shop, Pune. HPLC grade ammonium acetate and acetonitrile were obtained from Merck Specialties Private Limited, Mumbai, India. Syringe filters (0.22 μm), filter papers (0.45 μm) and ethylenediamine tetraacetic acid (EDTA) tubes were obtained from Merck Specialties Private Limited, Mumbai, India. Male Wistar Rats (200-220 gm) were purchased from Agharkar Research Institute, Pune
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