A simple, economic, selective, precise and accurate High-Performance liquid Chromatographic method used for the estimation of Adenosine in bulk drug. The mobile phase used was of Mixture of Acetonitrile and water in the proportion 5:95 respectively. This Mobile phase was allowed to flow at rate of 0.8ml/min. And this was found to give a sharp peak of Adenosine at a retention time of 3.78 min. Analysis of HPLC for Adenosine was carried out at a wavelength of 256 nm. Linear regression analysis data for the Calibration curve showed a good linear relationship, in concentration range of 50-100ppm and regression coefficient 0.991. The linear regression equation was Y=71258× the developed method was employed with a high degree of precision and accuracy for the analysis of adenosine. The inter and intraday variation was less than 2%. The mean recovery of the drug was 99.39%. The proposed method is simple, fast, accurate, and reproducible hence, it can be applied for routine quality control analysis of Adenosine.