The purpose of the Ames short-term bacterial reverse mutation experiment is to identify a variety of chemicals that can cause genetic harm that results in gene mutations. In this study, we investigated the mutagenic potential of 1-[[(3-hydroxy-1-adamantyl) amino] acetyl] pyralidine-(S)-2-carboxamide (Vildagliptin amide impurity) using Salmonella typhimurium and Escherichia coli tester strains. The evaluation utilized plate incorporation methods to assess point mutations at the histidine locus in four strains of Salmonella and at the tryptophan locus in E. coli WP2uvrA, both with and without a metabolic activation system (S9). A series of concentrations (15.8 to 5000 µg/plate) were tested, revealing no significant decreases in revertant colony counts or cytotoxicity, as evidenced by the integrity of the bacterial lawn and lack of precipitation. Confirmatory assays at varying concentrations indicated that treatment did not induce notable increases in revertant counts across the tester strains, further supporting the absence of mutagenic activity. Positive controls demonstrated expected responses, confirming the reliability of the test conditions and the functionality of the metabolic activation system. Overall, the findings suggest that Vildagliptin amide impurity does not exhibit mutagenic properties under the tested conditions.