ANALYTICAL METHOD DEVELOPMENT AND VALIDATIONFORTHEESTIMATIONOFTENOFOVIRN DISOPROXIL FUMARATE BY USING RP-HPLC

A simple, precise, and accurate Reverse Phase High-Performance Liquid Chromatographic (RP-HPLC) method was developed for the quantitative estimation of Tenofovir Disoproxil Fumarate (TDF) in tablet dosage form. The separation was carried out using an ODS C18 column (250 × 4.6 mm, 5 µm) with a mobile phase consisting of Acetonitrile, Methanol, and Water in the ratio of 5:50:45 (v/v/v), along with 1 mL of 0.1% Ortho Phosphoric Acid (OPA). The flow rate was maintained at 1.0 mL/min with UV detection at 260 nm and an injection volume of 20 µL. The retention time of TDF was found to be approximately 5.92 minutes. The method showed excellent linearity in the concentration range of 0.4–0.6 mg/mL (80%–120%) with a correlation coefficient (R²) of 0.9999. The assay of the marketed formulation demonstrated 99.12% of the labeled amount of TDF, falling within the ICH acceptance range of 97.0%–102.0%. System suitability parameters such as theoretical plate count, tailing factor, and %RSD were within acceptable limits, confirming method reliability. Hence, the proposed RP-HPLC method is sensitive, accurate, and reproducible, and can be successfully applied for routine quality control analysis of Tenofovir Disoproxil Fumarate in pharmaceutical dosage forms.