The chase to improve the quality of life has stimulated desirable changes in research to design and developa new drug and enhanceits safety and effectiveness. Thus, there is a gradualrise in demand to develop susceptible and specific analytical techniques for newly developed drugs. Thus, analystsare striving very hard to develop new and efficientanalytical methods to achieve these targets. Semaglutide was approved by United states Food and Drug administration(USFDA). It comes under antidiabetic agents; it acts by improves the efficiency of incretin function by activating GLP-1 receptors. This review article represents the collection and discussion of various analytical methods available in the literature for the estimation of the Semaglutide in pharmaceutical formulations and biological samples consisting of UV, HPLC, UPLC and hyphenated techniques such as LC-MS, LC-MS/MS. Moreover, we discuss about Semaglutide chemical structure, mechanism of action, and pharmacodynamics/pharmacokinetics properties. The present review can be effectively exploredto conduct future analytical investigation for the estimation of Semaglutide. Keywords: Semaglutide, Analytical Methods, RP-HPLC, UPLC, LC-MS/MS.