An accurate, sensitive high performance liquid chromatography method for the simultaneous determination of Tenofovir disoproxil Fumarate (TNF) and Emitricitabine (EMT) in dosage form was developed and validated as per ICH Guidelines. The separation and quantification were achieved on Agilent C18 column using Methanol – Phosphate buffer7.0 (65:35 v/v) as mobile phase at a flow rate of 1 ml / min with UV detection at 260nm for both analytes. Emitricitabine and Tenofovir disoproxil Fumarate were eluted at 4.7 min and 5.8 min with asymmetry of 1.27 and 1.22 respectively. Theoretical plates were found to be more than 9000 and the linearity ranges were 5- 70 μg / ml and 5 -70 μg/ml, respectively for TNF and EMT. The intra and inter – day RSD’s were in the range of 0.6 – 0.9% and 1.2 – 1.6% for TNF and EMT. The LOD and LOQ values for EMT and TNF were found to be 0.054 μg/ml, 0.165 μg/ml and 0.063μg/ml,0.192μg/ml respectively. The recovery studies were found to be 98.7±0.6 %, 100.5±1.1, 100.3±0.5 and 99.8±1.1, 101.2±0.8, 101.2±0.3 for LQC, MQC, HQC samples of TNF and EMT, respectively. The assay values were in the range of 98 – 102 %.