A simple, sensitive and accurate spectrophotometric method was developed in ultraviolet region for the estimation of Nebivolol (NEB) in pure drug, pharmaceutical formulation. Linear response obtained was in the concentration range of 5-45 µg/ml with correlation coefficient of 0.9995, 0.9989 in solvent and plasma respectively. Excellent recovery proved that the method was sufficiently accurate. There is no interference from any common pharmaceutical additives and diluents. Results of the analysis were validated by recovery studies according to ICH Q2(R1) guidelines and the same method is applied for estimation of NEB from plasma and these results were validated according to USFDA guidelines for bioanalytical method.