Stability indicating method development and validation of semaglutide by RP-HPLC in pharmaceutical substance and pharmaceutical product

A New specific, economic and selective, accurate, precise and robust Reverse Phase High Performance Liquid Chromatography was developed for the quantification of Semaglutide in pharmaceutical substance and product. Chromatographic separation was achieved by C18 column (Azilent C18 150x 4.6, 5pm) is used as stationary phase and 0.01N Potassium dihyrogen ortho phosphate: Acetonitrile (50:50) used as a mobile phase at a flow rate of 1.0 mL/min and monitored at 230nm. The run time was 5min. The retention time of Semaglutide was found to be 2.222min. To fulfil the International Conference on Harmonisation requirements developed method was validated. Validation parameters include system suitability, specificity, linearity, accuracy, precision, intermediate precision and robustness. Calibration curve was linear over the concentration range of 7-42 pg/ml. From regression analysis relative correlation coefficient (R2) value was found to be 0.999. Accuracy and precision results were within the limits. LOD and LOQ were found to be 0.007 pg/ml and 0.022 pg/ml respectively. Semaglutide was subjected to the stress conditions like acidic, basic, oxidative, photolytic and thermal conditions. In acidic, alkaline and peroxide stress conditions. In acidic, alakaline and oxidative conditions degradant peaks were observed but there is no interference with semaglutide peak in all accelerated conditions, hence proving the stability indicating and specific nature of method. Assay result was found to be 99.99% W/W, thus it is proving that the method can also be applied for the estimation of Semaglutide in pharmaceutical drug product.