RP-HPLC Method Development and Validation for Simultaneous Estimation of Emtricitabine and Tenofovir in Pure and Pharmaceutical Dosage Form

Abstract

A simple, precise, accurate RP-HPLC method with PDA detector has been developed and subsequently validated for the simultaneous estimation of Emtricitabine and Tenofovir in pure and pharmaceutical dosage form. The estimation was carried out on Hyperclone C18 (250mm x 4.6mm i.d; particle size 5µm) column with a mixture of potassium di hydrogen phosphate buffer (pH-2.5 adjusted with orthophosphoric acid) and methanol in the ratio of 35:65 v/v as mobile phase at flow rate 1ml/min with UV detection performed at 254 nm. The retention times of Emtricitabine and Tenofovir were 2.580 min & 3.711 min respectively. The concentration range was found to be linear 30-70 µg/ml for Emtricitabine and 45-105 µg/ml for Tenofovir. The correlation coefficient (r2) was found to be 0.999 for both the drugs. The limit of detection was found to be 2.92 and 3.05 and the limit of quantification was found to be 3.05µg/ml and 10.07µg/ml for Emtricitabine and Tenofovir. The %RSD values were less than 2 for both the drugs. The assay of Emtricitabine and Tenofovir was found to be 99.77% and 100.12% respectively. The method was validated for linearity, accuracy, precision, robustness, LOD, LOQ as per ICH guidelines. The developed method was successfully used for the quantitative analysis of commercial available dosage form.