An accurate, precise, robust and stability-indicating RP-HPLC method was developed for the estimation of Semaglutide using QbD approach. After conducting several trials using CCD method, one desirable method was optimized. Stationary phase selected was Kromasil C18 (250×4.6 mm, 5 µm) and potassium dihydrogen orthophosphate (pH 2) and Methanol used as mobile phase in the ratio of 61.2: 38.8. Detection was carried out at the wavelength 230nm. Flow rate selected for separation was 0.98ml/min and the temperature of 29.150C. The retention time was found to be 2.518 at the run time of 5min. The developed method was subjected to validation as per ICH guidelines. Semaglutide linear in the concentration range of 0-9µg/ml and the regression coefficient was found to be 0.999. The developed method was accurate, precise and robust. When a sample solution subjected to stress conditions like acid hydrolysis, neutral hydrolysis, photodegradation and thermal degradation, no degradation products were observed. In basic degradation studies two degradation products and oxidative conditions one degradation product was observed. From stress studies, we can observe that the degradation of Semaglutide was less than 10 in all conditions, and there is no impact of degradation products on system suitability parameters. The peak was homogenous I all conditions, thus proving the stability-indicating nature of the method. This method can be applied for the determination of Semaglutide in the drug substance.