Genotoxic effects of commercial sample of ponceau 4R-based food colorant in micronucleus assay on human whole blood culture

BACKGROUND: Genetic safety assessments of food additives have traditionally been conducted using highly purified substances. In the Russian Federation, according to the requirements of the Customs Union Technical Regulation, genotoxicity testing of approved food colorants is not mandatory. The regulation is limited to determining the content of the main colorant component and select constituents. However, this approach is insufficient, as it does not account for the possible presence of toxic and genotoxic impurities in food colorants.AIM: To evaluate the genetic safety of a commercially available Ponceau 4R (E124)-based food colorant using the micronucleus assay on human whole blood culture with cytokinesis block, both in the presence and absence of a metabolic activation system.METHODS: The Ponceau 4R-based colorant was purchased in a retail store. The cells from a healthy donor were cultured under cytokinesis-block conditions with and without S9 rat liver metabolic activation, exposed to the colorant at concentrations ranging from 0 to 2 mg/mL. Cytome assay was performed using an extended micronucleus assay protocol. Statistical analysis was conducted using the χ2 test and the Mann-Whitney U test.RESULTS: A statistically significant increase in the frequency of cells with genetic damage was observed, following a U-shaped dose-response pattern. Without metabolic activation, significant effects were found at concentrations of 0.0000256, 0.00064, and 0.4 mg/mL; with S9 activation, at concentrations of 0.0000256, 0.000128, and 0.016 mg/mL. In addition, in the presence of the S9 fraction, an increased frequency of trinucleated cells, stimulation of mitotic activity, and suppression of apoptosis were also observed.CONCLUSION: Genotoxic effects of the Ponceau 4R-based food colorant obtained from the retail market were detected at or below the acceptable daily intake level for humans. The proposed approach may serve as a foundation for the development of a system for genetic safety assessment of food colorants and additives

Micronucleus · Micronucleus test · Biochemical Analysis and Sensing Techniques · Chemistry · Consumer Attitudes and Food Labeling · Dye analysis and toxicity · Food Science · Organic Chemistry