A sensitive, accurate and precise stability-indicating high performance liquid chromatography (HPLC) method was developed for determination of odevixibat in pure form and pharmaceutical formulation. In this study, comprehensive stress testing of odevixibat has been carried out according to ICH guidelines Q1A (R2). Odevixibate was subjected to forced degradation studies under hydrolytic (acid and base), oxidative, photolysis, and thermal stress conditions. The chromatographic separation was performed using a Phenomenex C8 column, flow rate of 1.0 mL/min, and UV detection at 224 nm. The optimized HPLC system was achieved using mobile phase composition containing methanol:water (85:15, v/v). The proposed method showed well separation between the drugs and its degradation products and had a good accuracy. The proposed method showed well separation between the drugs and their degradation products and had a good accuracy. The method was linear with 5-50 ng mL-1 for odevixibat. The method showed to be linear (r2 > 0.999), precise (RSD < 0.350%), accurate (recovery of 99.95% for odevixibat), specific and robust. LOD and LOQ values were 1.09 ng mL-1 and 3.60 ng mL-1 for odevixibat. The method was validated according to ICH guidelines and applied for determination of the cited drugs in its pharmaceutical formulation.