Development and Validation of a Rp-hplc Method for Simultaneous Estimation of Lamivudine, Tenofovir Disoproxil Fumarate and Efavirenz in a Combined Tablet Dosage Form

Objective: Development of an accurate, simple, precise and rapid method for estimating Tenofovir disoproxil fumarate, Lamivudine and Efavirenz simultaneously, in a combined tablet form. Methods: The method uses Reverse Phase High Performance Liquid Chromatography (RP-HPLC). Symmetry C18 (4.6 x 100 mm, 3.5 μm) column operated with a mixture of phosphate buffer of pH 4.0 with ortho phosphoric acid and Acetonitrile (42:58) as mobile phase was found to be suitable for the simultaneous estimation. The flow rate was maintained at 0.5 mL/min. Detection was carried out at 254 nm using a UV detector. Results: The total run time was less than 11 minutes with the retention times of 2.2 min, 3.3 min and 10.8 min respectively for Lamivudine, Tenofovir disoproxil fumarate and Efavirenz. Conclusion: Validation of the method was performed for precision, accuracy, linearity, ruggedness, specificity and sensitivity to conform to the ICH guidelines for validation of an analytical method.