Since the mid-1950s, the volatile structure of butter oil and butter were researched, and an exhaustive list of elements has been collected. Diacetyl is an aromatic popular synthetic fragrance that gives food a buttery taste used in ice cream, snacks and potting with butter, strawberry, caramel, or cheese flavor. The chromosomal aberrations and micronuclei are commonly used biomarkers of chromosomal damage, genome stability, and cancer risk assessment.In vivo trials are still important to assess the genetic toxicology of chemical products such as industrial chemicals, pharmaceuticals, and food additives. This study aimed at assessing the potential genotoxic effect of diacetyl and butter flavors on swiss albino mice using alterations in liver function enzymes, micronucleus (MN), and chromosomal aberrations (CA) assays. The results showed that exposure of swiss albino mice malesto diacetyl and butter flavors induced (CA) and (MN) in a statistically highly significant manner compared to the control. Meanwhile, the biochemical analysis revealed that these substances caused an exceptional rise in liver function enzymes (AST, ALT, and ALP) activity in serum of treated experimental animals.In conclusion, both tested compounds have increased the chromosomal aberration, micronucleus test, and serum levels of liver function enzymes indicating their high potential of being cytotoxic and genotoxic materials.