Bacterial Reverse Mutation Test of 2-(3-hydroxyadamantan-1-yl)-1-iminohexahydro-pyrrolo [1,2-a] pyrazin-4-one (Vildagliptin cyclic amidine impurity) using Salmonella Typhimurium and Escherichia Coli

The Ames Salmonella/microsome mutagenicity assay (Salmonella test; Ames test) is a short-term bacterial reverse mutation assay specifically designed to detect a wide range of chemical substances that can produce genetic damage that leads to gene mutations. In this study, we are going to investigate the mutagenic potential of 2-(3-hydroxy-adamantan1-yl)-1-iminohexahydro-pyrrolo[1,2-a] pyrazin-4-one (Vildagliptin cyclic amidine impurity) using Salmonella typhimurium and Escherichia coli tester strains. The evaluation was conducted to assess point mutations at the histidine locus in four strains of Salmonella typhimurium and the tryptophan locus in E. coli WP2uvrA. The experiment employed the plate incorporation method, both with and without a metabolic activation system (S9). Cytotoxicity was assessed by measuring the thinning of the bacterial lawn and the reduction in revertant colony counts compared to vehicle controls. Results indicated no significant cytotoxic effects or mutagenic activity across tested concentrations (15.8 to 5000 µg/plate) in both the presence (10% and 20%) and absence of metabolic activation. The findings demonstrated that Vildagliptin cyclic amidine impurity did not induce point mutations in the selected bacterial strains, highlighting the lack of mutagenic potential of the compound within the tested parameters.