A reverse phase High pressure liquid chromatography (HPLC) method to determine puerarin in Pueraria tuberose was developed and described. The method involves separation of active compound using mobile phase consisting of acetonitrile: water: orthophosporic acid in the ratio of 32:67:01 and detecting the chromatogram at 280 nm using photodiode array detector. The sensitivity of the method was observed as 1.0 mg and linearity was observed between 1.0 mg to 8.0mg. The puerarin content of 3.02% w/w was found in crude P.tuberosa The proposed method in precise, sensitive, reproducible and easy to perform which can be used for detection, monitoring and quantification of puerarin in P . tuberose.