Box-Behnken Design Assisted AQbD Approach for the Optimization and Quality Assessment of Ensitrelvir in Bulk and Dosage forms by RP-HPLC

ABSTRACT: Ensitrelvir, is an oral SARS-CoV-2 3CL protease inhibitor that was approved in Japan to treat SARS-CoV-2 infections. This paper describes the AQbD approach and Box Behnken Design assisted development of a HPLC method and its validation for Ensitrelvir in bulk and dosage form. The three independent variables of the RP-HPLC method were flow rate, organic ratio in mobile phase and runtime and the responses retention time and tailing factor were taken as dependent variables. The study utilized a PLATSIL C18-EP column (4.6 x 250mm, 5µm) the chromatographic conditions were optimized using Acetonitrile: Triethylamine pH: 4 (60:40 mL) as the mobile phase, 1 mL/min as flow rate with a Rt of 9.609 min, at a λ max of 228 nm. The devised technique was found to be linear with a serial dilution of 10–50 μg/ml with (r2) of 0.991. The tailing factor (TF) and theoretical plates (N) were 1.28 and 4883 results indicated the system suitability test, respectively. The precision for Intraday and Interday were determined and % RSD was observed to be 1.6 and 0.9 %. The robustness values were below 2%. No other coeluting peaks were found with the Ensitrelvir peak, according to the chromatographic peak purity data. According to ICH specifications, the parameters for method validation were within the permissible range.