Analytical Quality by Design‐Based Stability‐Indicating Ultra‐High Performance Liquid Chromatography Method for Estimation of Acoramidis Hydrochloride in Bulk and Tablet Dosage Forms

ABSTRACT Acoramidis hydrochloride is a novel transthyretin stabilizer recently approved for the treatment of transthyretin amyloid cardiomyopathy. This study presents the development and validation of a novel, robust, and stability‐indicating reversed‐phase ultra‐high performance liquid chromatography (RP‐UHPLC) method for the quantification of acoramidis hydrochloride in both bulk and pharmaceutical dosage forms. Employing Analytical Quality by Design (AQbD) principles, method optimization was achieved using a phenyl column (50 × 1.0 mm, 1.7 µm) and an isocratic mobile phase comprising 0.1% formic acid and acetonitrile (50:50, v/v) at a flow rate of 0.5 mL/min. The injection volume was 5 µL, and the analyte was detected at 228 nm, with a total run time of 2.5 min. The method validation was carried out in compliance with ICH Q2(R1) guidelines, demonstrated acceptable specificity, precision (%RSD < 2), linearity ( r 2 > 0.9998) over the 5.0–1500.0 µg/mL concentration range, as well as satisfactory accuracy (99.49–100.24% recovery), sensitivity (LOD: 1.6 µg/mL; LOQ: 5.0 µg/mL), and robustness. To assess the method's specificity, forced degradation studies carried out under different stress conditions, such as acidic, alkaline, oxidative, thermal, photolytic, and neutral conditions, confirmed the method's ability to selectively quantify acoramidis hydrochloride in the presence of degradation products. Mass balance values exceeded 95% in all conditions, and peak purity tests passed under all stress conditions. The robustness of the method was assessed using an analytical quality by design approach by introducing deliberate variations in critical parameters, including the flow rate of the mobile phase, the column temperature, and the organic ratio of the mobile phase. Therefore, this developed method, with a short run time of 2.5 min, is reliable and suitable for routine quality control and stability testing of acoramidis hydrochloride in pharmaceutical research and development and quality control departments.