eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2025-09-30 22 3 1162 1175 10.13005/bbra/3431 56207 article Madhu Reddemma 1 Pratyusha Valligatla 1 Sivagami Bojan 1 Sunil Kumar Ellampati 1 Mounika Tummalapalli 2 Charumathi Salva 3 Department of Pharmaceutical Chemistry, Seven Hills College of Pharmacy (Autonomous), Andhrapradesh, India. Department of Pharmaceutical Chemistry¸ Acharya Nagarjuna University, Guntur, India Department of Pharmaceutical Analysis, RBVRR Womens College of Pharmacy, Telangana, India A sensitive and robust UPLC method was developed and validated for the simultaneous analysis of bempedoic acid and ezetimibe in drug substance and drug product. Chromatographic conditions were accomplished using a UPLC (Acquity) CSH C₁₈ column (100 × 2.1 mm, 1.7 μm) under isocratic conditions, utilizing a movable phase comprising of acetonitrile and heptane sulphonic acid buffer (pH 2.5) /OPA in a 5:95 v/v ratio. The absorption wavelength was set at 232 nm, with a flow rate of 0.5 mL/min. International Council for Harmonisation Q2(R1) standards were used for method optimization and included stress studies. The retention times for bempedoic acid and ezetimibe were 0.365 and 1.326 minutes, respectively. The method demonstrated a regression coefficient (R² > 0.999), ruggedness (% RSD < 2%), recovery range (between 98–102%) and robustness. The LOD was 0.54 μg/mL for bempedoic acid and 0.03 μg/mL for ezetimibe. Both compounds exhibited sensitivity to acidic, oxidative, and thermal degradation, confirming the stability-indicating method of the compounds. Overall, the developed UPLC method is precise, accurate, specific, and suitable for quality analysis and stability assessment of bempedoic acid and ezetimibe in both drug substances and drug products. https://www.biotech-asia.org/vol22no3/quantification-of-bempedoic-acid-and-ezetimibe-with-comprehensive-stability-assessment-in-drug-substance-and-drug-product-by-uplc/ Bempedoic Acid; Ezetimibe; ICH; Method Development; Stability Assessment; Simultaneous Quantification; UPLC; Validation