Daunorubicin is an anthracycline antibiotic it used to treat cancer. A simple, sensitive and specific reverse phase high performance liquid chromatographic (RP HPLC) method for the determination of Daunorubicin was developed and validated by using Cosmosil C18 column (250 mm x 4.6 mm, 5pm) using a mixture of Water(0.1% OPA) and Acetonitrile in the ratio 05:95 (v/v) as a mobile phase This was found to give sharp peak of Daunorubicin at a retention time of 4.3167 min. RP HPLC analysis of Daunorubicin was carried out Detection of wavelength by UV detector at 230 nm with a flow rate of 1ml/min. The applied RP HPLC method allowed Daunorubicin with good linearity (r2 = 0.999) in the studied concentration range. Limit of detection and limit of quantification were found to be 0.21µg/ml and 0.66µg/ml respectively. The method was validated as per the International Conference on Harmonization (ICH) guidelines. The developed method was employed with a high degree of precision and accuracy limit of quantification, limit of detection. The method was validated for accuracy, precision, robustness. The precision, accuracy, repeatability, short retention time and composition of the mobile phase indicated that this method is better for the /11 15:13 quantification of Daunorubicin,