Neonicotinoids, which are widely used worldwide, including in Turkey, are an insecticide group that are synthetic derivatives of nicotine. Recently, they have attracted attention due to their toxic effects on non-target organisms, especially bees. Numerous studies have shown that neonicotinoids have been found in detectable levels in the environment and cause various undesirable effects on living organisms, including humans and other mammals. In this study, the possible toxic effects of imidacloprid and acetamiprid, commonly used neonicotinoids, are investigated by their pure forms and commercial formulations on HT-29 cells with individual and combined exposures. According to our results, imidacloprid and acetamiprid induced cytotoxicity by caspasemediated apoptosis, mitochondrial membrane depolarization, DNA damage, and oxidative stress under these experimental conditions. It is worth mentioning low doses of DNA damage, mixture exposure causes toxic effects at lower concentrations than individual exposure, and formulation groups are at the forefront of toxicity formation, though this varies depending on the parameters.