Introduction: Analytical methods like Ultraviolet-visible spectroscopy and High-Performance Liquid Chromatography play a critical role in equivalence and risk assessment and management. Nucleotide reverse transcriptase inhibitors tenofovir disoproxil fumarate (TDF) and Emtricitabine (ETB) is clinically a potent combination against HIV and hepatitis B virus devoid of short-term irritating toxicity. Objective: The present investigation deals with the development and validation of new UV spectrophotometric Vierordt__ampersandsignrsquo;s and Reverse Phase Ultra-Fast Liquid Chromatography (RP-UFLC) methods for the simultaneous estimation of TDF and ETB in bulk and tablet dosage form as per the International Council for Harmonization (ICH) guidelines. Methods: Vierordt__ampersandsignrsquo;s method was developed on Shimadzu UV-1800 UV/Visible Spectrophotometer. Reverse phase ultra-fast liquid chromatography was developed on Shimadzu UPLC, X-Bridge C18 column equipped with a photodiode array detector. Mobile phase composed of water: methanol 62:38. The effluent was pass through the column 1.0 ml/min at 30o C and the response was recorded at 254 nm. Results: Absorption maxima of TDF and ETB were obtained at 260 nm and 281 nm respectively. The RSD of all validation parameters was lesser than 2 % indicated the accuracy of the Vierordt__ampersandsignrsquo;s method. In UFLC, the retention time of ETB and TDB were found to be at 3.371 and 6.850 min respectively. Purity angle and purity threshold of both peaks showed spectral homogeneity over all the peak region indicated its peak purity. Conclusion: The proposed Vierordt__ampersandsignrsquo;s and RP-UFLC methods found to be sensitive, rapid, accurate and economical and it can be employed for simultaneous estimation of ETB and TDB in bulk drugs and formulations.