Quality by Design Approach in HPLC Method Development and Validation of Baloxavir Marboxil in Pharmaceutical Dosage Form

Abstract

A simple, robust, and validated RP-HPLC method was developed for the quantitative estimation of Baloxavir in bulk and pharmaceutical dosage forms, as per ICH Q2(R1) guidelines. Chromatographic separation was achieved on a PLATISIL C18 column (4.6×250 mm, 5 µm) using a mobile phase of methanol and KH2PO4 buffer (pH 4.5) in a 60:40 v/v ratio, at a flow rate of 1 mL/min, with UV detection at 247 nm. System suitability parameters, including theoretical plates (>2000) and tailing factor (<2), complied with acceptance criteria, confirming method reliability. The method exhibited excellent linearity in the range of 10–50 µg/mL with a correlation coefficient (R²) of 0.9998. Precision studies yielded %RSD < 1%, while accuracy tests showed recoveries within 98–102%. Sensitivity studies revealed a LOD of 0.37 ppm and LOQ of 1.25 ppm, indicating high detection capability. Robustness and ruggedness evaluations confirmed the stability of the method under small, deliberate changes in chromatographic conditions. Overall, the validated RP-HPLC method is specific, precise, accurate, and reproducible, making it highly suitable for routine quality control, stability assessment, and regulatory analysis of Baloxavir formulations in the pharmaceutical industry.