The present study was aimed to develop a novel, simple, accurate, precise, robust and economic UV spectrophotometric method for the detection and quantification of vancomycin in bulk and its formulation. Quantification was carried at absorption maxima 280.50 nm using 0.01M Sodium hydroxide as diluent. The developed method was validated with accordance to ICH Q2 (R1) guidelines. Beer’s law was obeyed in the concentration range of 40-240 μg/mL with correlation coefficient of 0.9991. The sensitivity was checked as the limit of detection and limit of quantification which were found to be 1.6 and 4.8 μg/mL respectively. Further studies of the accuracy and precision of the proposed method were performed using standard addition method; the mean quantitative recovery of such studies were found to be in the range 99.39 ± 0.41 to 101.24 ± 0.89 with RSD ≤ 2.0%. From the results it can be concluded that the developed method was specific, selective, precise and robust. This method could be successfully applied for analysis of vancomycin in its formulation.