Stability-Indicating RP-HPLC Method for Determination of Ursodeoxycholic Acid in Tablet Dosage Form

Abstract

The aim of the present study is to develop and validate a novel, simple, selective and sensitive stability indicating reverse phase HPLC method for the determination of ursodeoxycholic acid (UDCA) in tablet dosage form after being subjected to different stress conditions, such as hydrolysis (0.1 N HCl and 0.1 N NaOH), oxidation (30 % H₂O₂), heat (80 ºC for 48 h) and photolysis (UV, 254 and 366 nm). The validation studies were carried out as per the International Conference on Harmonization (ICH) and United States Pharmacopoeia (USP) guidelines. An isocratic HPLC method was developed to separate ursodeoxycholic acid from the degradation products, using a BDS Hypersil C₈ column (thermo, 250 mm × 4.6 mm, 5 μ) with an isocratic mobile phase comprising of methanol, water and phosphoric acid (77:23:0.6 v/v).The flow rate was 1.0 mL/min and was carried out in refractive index detector. Retention time for ursodeoxycholic acid was about 3 min. A linear response was observed in the range of 240-360 μg/mL, (r² = 0.995) for the drug. The drug was decomposed in acid, base, 30 % H₂O₂ and in heat but was found to stable in photolytic stresses. The method was validated in terms of linearity, precision, accuracy, specificity, limit of detection and quantitation and robustness. The procedure was found to be specific, linear, precise (including intra and inter day precision), accurate and robust. Applicability of the method has been illustrated performing the assay of the drug in the tablet.