Aspartame and acesulfame potassium are among the best known food sweeteners. Despite numerous studies showing the safety of these supplements, in recent years the attention of researchers has been focused on the negative side effects of their exposure, which can lead to the development of non-communicable diseases. The aim of this investigation was to study the nature of the impact (reversible or irreversible) of aspartame and acesulfame potassium on weight gain, as well as cytological, cytogenetic and metabolic parameters of onion roots. Roots were pre-treated with 0.5 and 1 g/l aqueous solutions of sweeteners, and then they were transferred to water for the purpose of further regenerative germination. The consequences of the toxic effect of aspartame on the weight gain of onion roots were minimal compared to the control. This indicator significantly and dose-dependently decreased to 38% in the samples with acesulfame potassium, which shows the presence of a delayed effect after exposure to this sweetener. In the experimental groups with the maximum content of sweeteners, a lower mitotic index (p≤0.05) in the meristem cells was also maintained compared to the control. The results obtained indicated an irreversible mitotoxic effect of these compounds. However, an increase in the frequency of mitosis pathologies found in the experimental samples due to pre-treatment with sweeteners was completely leveled after regenerative germination. Therefore, these genotoxic disorders were reversible. To assess the processes of lipid oxidation of root tissues, the concentration of malondialdehyde was measured. The effect of reducing the level of this indicator in all experimental samples compared with the control turned out to be irreversible, although not dose-dependent. The results obtained can be used in the development of formulations for adding food sweeteners to food products to reduce the risk of delayed toxic effects.