Establishment of health food safety evaluation method-In vivo genotoxicity evaluation by micronucleus assay

In recent years, various factors such as shifts in health consciousness, changes in lifestyle habits, increased focus on dietary intake, the promotion of alternative medical concepts, and advancements in life sciences and technology have propelled the robust growth of the health food industry. An increasing number of reports suggest that "functional foods," containing beneficial functional components, may enhance short-term well-being and are increasingly regarded as healthful. Building upon these findings, this study aims to enhance the production capacity of high-value agricultural products. However, for quality assurance, toxicity assessment of functional foods is imperative. There is a growing necessity to employ in vivo genotoxicity assays to evaluate the carcinogenic potential of these products. In this study, we tried to establish another in vivo genotoxicity evaluation platform via acridine orange induction. According to all results in this study, during the experiment, there were no statistically significant differences in BW between the negative control group and the normal control group (p > 0.05). Moreover, the clinical observations of the experimental mice showed that all mice survived until the end of the experiment, and no abnormal clinical symptoms were observed. The mouse’s food consumption in each group were monitored during the experiments. The food consumption of mice was recorded daily after the experiment. During the experiments, there were no significant difference between two groups on the food consumption. The percentage of RETs/1,000 RBCs in the negative control group was significantly decrease than the normal control group (p < 0.001). RETs/1,000 RBCs (‰) in ICR mice (n = 5/group) in the negative control group and the normal control group at 48^th and 72^th hours-experiment were respectively 18 ± 4.6 / 45.6 ± 4.0 (48^th hours-experiment) and 19 ± 4.8 / 47.6 ± 4.8 (72^th hours-experiment). The percentage of Mn-RETs/1,000 RETs in the negative control group was significantly increase than the normal control group (p < 0.001). Mn-RETs/1,000 RETs (‰) in ICR mice (n = 5/group) in the negative control group and the normal control group were 10.6 ± 2.1 / 0.4 ± 0.5 and 8.2 ± 1.8 / 0.4 ± 0.5, respectively. Taken all results of RET/RBCs (‰) and MN-RET/RETs (‰) together, the genotoxicity in the negative control group has been successfully induced. In light of these requirements, we propose utilizing the mammalian erythrocyte micronucleus assay to detect damage induced by test substances to erythroblast chromosomes. This study endeavors to establish a sensitive and stable method for quantifying micronuclei formation in erythrocytes in vivo. The resulting data will facilitate the evaluation of potential toxicity associated with agricultural functional products in the future.