Eco-optimized RP-HPLC method for chiral tazobactam and piperacillin drugs: integration with Box–Behnken design and multi-sustainability tools

Piperacillin-tazobactam (PIP-TAZ) is an effective antimicrobial agent, indicated primarily for severe infections resulting from susceptible organisms, and it has undisputed importance within antimicrobial therapy. PIP-TAZ combinations are widely used to treat severe infections caused by Gram-negative and Gram-positive bacteria, including those that produce β-lactamases. Our study involved the development of an environmentally friendly RP-HPLC method for the simultaneous determination of TAZ and PIP. In lieu of conducting numerous experiments, an efficient method to achieve the three major parameters of chromatography-pH, flow rate, and column temperature-was optimized with the help of a Box-Behnken Design (BBD) to minimize variability. This approach streamlines the experimental process and enhances results reliability. By carefully controlling these parameters, researchers can achieve more consistent and reproducible chromatographic analyses. This research addresses the impact that these variables have on such critical responses as retention times for TAZ and PIP and their resolution. The identification of optimal conditions was based on desirable and overlay plots. The samples were dissolved in ethanol, water, and tetrabutylammonium hydroxide in the mobile phase and separated through chromatography (58.5:40:1.5, v/v/v), and the pH was maintained at 3.5 with phosphoric acid. The method used UV detection at 230 nm, an injection volume of 20 μL, and a column temperature of 30°C. It was run at a flow rate of 1.0 mL/min. It used a Symmetry Shield RP18 column for 100-micron pore size, 5-micron particle size, and 4.6-mm by 250-mm dimensions. A comparison of carbon footprint reduction index (CaFRI), analytical greenness (AGREE), analytical eco-scales, green analytical procedure index (GAPI), modified GAPI (MoGAPI), and analytical method volume intensity (AMVI), which authenticates the accuracy, precision, and robustness of the method, was made in conjunction with a lesser environmental impact.