QUALITY-BY-DESIGN–DRIVEN DEVELOPMENT AND OPTIMIZATION OF RP-HPLC METHOD FOR CONCURRENT ESTIMATION OF AMOXICILLIN, VONOPRAZAN, AND CLARITHROMYCIN IN A SYNTHETIC MIXTURE.
A reverse-phase high-performance liquid chromatography (RP-HPLC) method, developed under the Analytical Quality by Design (AQbD) framework, was systematically optimised for the simultaneous quantificationof amoxicillin trihydrate (AMO), vonoprazan fumarate (VONO), and clarithromycin (CLA) in a synthetic mixture. Critical method parameters such as trifluoroacetic acid (%TFA) concentration, mobile phase ratio and columntemperature were optimised using a Central Composite Design (CCD) to achieve the desired chromatographicresponses, including optimal retention time, theoretical plate count, and minimal peak tailing. Chromatographicseparation was achieved on a C18 column (4.6 × 100 mm, 3 μm particle size) using a gradient elution program, withUV detection carried out at 210 nm. Method validation, performed in accordance with ICH Q2(R2) guidelines, confirmed excellent linearity (r² > 0.999), accuracy, precision, and robustness within the validated range. Thefindings confirm the method’s suitability for routine quality control and stability testing of multicomponent formulations. This study highlights the effectiveness of AQbD principles in developing a robust, reliable, andregulatory-compliant analytical method, thereby contributing to enhanced pharmaceutical method development andensuring consistent product quality.
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