An integrated framework to develop an efficient valid green (EVG) HPLC method for the assessment of antimicrobial pollutants with potential threats to human health in aquatic systems†

The persistence of antimicrobial drugs in aquatic environments has raised critical concerns about their possible impact on drinkable water quality and human health. The Nile River is experiencing water pollution owing to increasing discharges of highly contaminated home and industrial effluents and inadequate water management systems. Investigations of the presence of three antimicrobial agents, ciprofloxacin (CIP), sulfamethoxazole (SMZ), and albendazole (ALB), in the Egyptian aquatic system are recommended using a chromatographic method because of their reported existence in the African aquatic environment. In this study, an integrated framework, Efficient Valid Green (EVG), for analytical techniques is proposed and displayed via its radar chart. The EVG framework is achieved through three main pillars: efficiency, validation, and greenness. The proposed EVG-HPLC method was developed and optimized using the AQbD methodology via a face-centered composite (FCC) design by identifying the proper critical method parameters (CMPs) that influence critical quality attributes (CQAs). The method was fully validated according to ICH guidelines, including a factorial robustness study within concentration ranges of 1–100 μg mL⁻¹, 2–100 μg mL⁻¹, and 10–100 μg mL⁻¹ for CIP, SMZ, and ALB, respectively. The proposed method was evaluated in terms of greenness using AGREE (score 0.55) and ComplexGAPI metrics. The optimized chromatographic conditions included a C18 column and a mobile phase of water : acetonitrile : methanol in a ratio of 60 : 19 : 21, v/v/v, respectively, with an aqueous solution of pH 3.5 adjusted with phosphoric acid at a flow rate of 1.57 mL min⁻¹ at 285 nm. The raw water samples collected from Nile River freshwater at different locations were treated using Oasis® PRiME HLB cartridges with satisfactory recoveries for the three analytes (>90%), and the three drugs were detected using the proposed EVG-HPLC method.