A quality by design HPLC method for cephalosporin analysis in pharmaceuticals and water samples with environmental impact assessment.

The present study applied a combined analytical quality-by-design and green analytical chemistry approach to develop an HPLC method for the determination of four cephalosporin pharmaceuticals in both their formulations and water samples. These drugs include ceftriaxone, cefotaxime, ceftazidime and cefoperazone. A Box-Behnken experimental design was employed to optimize three chromatographic parameters: mobile phase composition, flow rate and buffer pH. The predicted optimal conditions involved using a mobile phase of acetonitrile and 0.04 M phosphate buffer at pH 6 in a 7:93 (v/v) ratio, pumped at 1.3 mL/min through a Nucleosil C18 (4.6 × 250 mm, 5 μm) column with UV detection at 240 nm. Under these optimum conditions, the developed HPLC method successfully separated the four drugs with good resolution in less than 6 min. Linearity was established across the concentration ranges of 5-300 µg/mL for ceftriaxone and cefotaxime, 5-400 µg/mL for ceftazidime and 5-100 µg/mL for cefoperazone. Furthermore, full validation of the method in terms of accuracy, precision, specificity and robustness was carried out as per ICH guidelines. The greenness profile of the optimized HPLC method was also evaluated using the Analytical GREEnness (AGREE) tool and found to be environmentally friendly with AGREE score of 0.75, making it a greener alternative for quality control and routine analysis of the investigated cephalosporins in their pharmaceutical formulations and tap water samples. Furthermore, the blueness assessment of the proposed HPLC method using the blue applicability grade index (BAGI) tool yielded a value of 77.5, indicating its high analytical practicality and substantial potential for routine analysis applications.