Trifluridine and Tipiracil Hydrochloride are anti-cancer drugs. A new HPLC method has been proposed for the simultaneous determination of Trifluridine and Tipiracil Hydrochloride tablets on Isocratic mode with 1.0 mL/min flow rate and a diluent consisting of a mixture of Acetonitrile: Water: Methanol (60:20:20v/v) was used for the study. And it used column Hypersil C18 (4.6×150mm, 5.0 µm) A Waters Model 2695 separation module HPLC system equipped with 996 PDA detector was used. The detection was done at 230 nm. The retention time were found to be 2.842 min, 3.878 min for Trifluridine and Tipiracil Hydrochloride respectively. Trifluridine showed linearity in the concentration range of 10-50 μg/mL with linear regression (R2 = 0.9999). The % RSD for precession and accuracy studies were found to be < 2%.The LOD and LOQ were found to be 0.6 µg/ml,12.8 and 1.9 & 38.90 µg/ml, respectively.. The proposed method was observed to be simple, economical and was validated according to the ICH guidelines for linearity, precision, accuracy and stability. This proposed method is highly sensitive, precise and accurate which reduces cost of analysis; hence recommended for routine quality analysis in laboratories