Forced degradation studies and development of RP-HPLC method for related substances of Tenofovir alafenamide in tablet dosage form

Abstract

Abstract The development and validation of an easy and rapid stability-indicating RP-HPLC technique for Tenofovir alafenamide and its impurities. In this newly developed method, chromatographic separation of Tenofovir alafenamide and its impurities was achieved on an Inertsil ODS-3V C18 (250 mm x 4.6 mm, 5 μ) column. The impurities were extracted by a mixture of Mobile Phase A: buffer solution: Acetonitrile: Purified water (20:02:78) and Mobile Phase B: Solvent Mixture and Purified water (75:25), The injection volume was 20 μL, the column temperature was 40°C, the flow rate was 1 mL/min, and the detection was carried out at 262 nm. The retention time of Tenofovir alafenamide, PMPA, PMPA anhydrate, Phenyl PMPA and PMPA isopropyl alaninate were 56.24, 5.69, 8.71, 25.89 and 37.13 respectively. The Correlation Coefficient within the acceptance criteria is not less than 0.999. The evaluated concentrations for Tenofovir alafenamide and its impurities were in the range of 0.5–7.5 ppm. The average recovery value was in the range of 90.2–113.9%. Tenofovir alafenamide’s LOD and LOQ were determined to be 0.1 μg/mL and 0.5 μg/mL respectively. Tenofovir alafenamide solution degradation behaviour was assessed using solution stability studies. A new, accurate, and precise method has been developed, evaluated, and elevated for the detection of impurities in Tenofovir alafenamide tablet dosage form. The results obtained from the validation study established that this method is specific, reliable, precise and effective. As a result, the proposed strategy can serve as an alternate method to determine related substances in routine analysis of tablet dosage form. GRAPHICAL ABSTRACT