Robust RP‐HPLC Method for Vadadustat: Development, Validation, Stability, and Green Metrics

ABSTRACT Chronic kidney disease is a degenerative disorder affecting millions worldwide, often leading to complications such as anemia. Vadadustat, a hypoxia‐inducible factor prolyl hydroxylase inhibitor, has been developed as an oral treatment to stimulate erythropoiesis. Environmentally sustainable, accurate, and reliable analytical methods are essential for regulatory compliance and quality control. The present study focuses on the development, validation, and greenness assessment of a stability‐indicating reverse‐phase high‐performance liquid chromatography method for the quantification of vadadustat in its pure form and synthetic mixture. The validated RP‐HPLC method was developed using a Kromasil C18 (250 × 4.6 mm i.d., 5 µm) column, with Ultraviolet detection at 317 nm, and a mobile phase comprising of 6 mM ammonium acetate buffer (pH 3.2) and acetonitrile (30:70 v/v) at a flow rate of 1.0 mL/min. Validation parameters including accuracy, precision, linearity, detection and quantification limits, and robustness, were evaluated according to international guidelines. Stress degradation studies were conducted under acidic, oxidative, basic, photolytic, and thermal conditions. The method showed a retention time of 3.9 min and effective separation of degradation products. Vadadustat degraded significantly under acidic, alkaline, and oxidative conditions but remained stable under photolytic and thermal stress. The method demonstrated excellent linearity (correlation coefficient = 0.9992), along with high precision and robustness. Environmental and practical assessments indicated favorable performance using various green chemistry evaluation tools. The developed RP‐HPLC method is stability‐indicating, precise, and robust with demonstrated environmental sustainability and practical applicability. Suitable for routine quality control and pharmaceutical analysis of vadadustat in dosage forms.