Evaluation of mutagenic activity of the food dye Ponceau 4R in a micronuclear test in mice

Introduction. Food monoazo dye E124 Ponceau 4R is used in the food, pharmaceutical, and cosmetic industries. An analysis of the available data on the evaluation of the genetic safety of E124 based on the main marker events of genotoxicity revealed their inconsistency, possibly associated with quantitative and/or qualitative differences in the studied samples for impurities.

The aim of the work is to evaluate the mutagenic activity in the micronucleus test in mice of several commercial samples of E124 present on the domestic market.

Materials and methods. Three samples of E124 from different manufacturers (India) were studied. To assess the identity of Ponceau 4R substances, there was used method of MIR-spectroscopy — universal ART technique. Aqueous dye solutions were administered to the stomach of male CBA × C57Bl6/j) mice at doses of 125 to 2000 mg/kg twice with an interval of 24 hours and preparations were prepared 24 hours after the last injection. To assess the frequency of polychromatophilic erythrocytes (PCE) with micronuclei (MN), 4000 PCE were analyzed, the proportion of PCE among all erythrocytes — 500 cells per animal.

Results. Ponceau 4 R was determined in all samples. All E124 samples caused an increase in the frequency of PCE with MN (p < 0.05–0.001, T test) after exposure at all levels studied in a linear dose-dependent manner. The proportion of PCE from all erythrocytes did not change.

Limitations. The data obtained do not allow determining the mechanism of the genotoxic action of the test substance.

Conclusion. An analysis of the frequency of MN PCEs in the bone marrow in mice after a double injection at doses of 125–2000 mg/kg revealed the genotoxicity of three samples Ponceau 4R.

Compliance with ethical standards. The study was approved by the local ethical committee of the Research Institute of EDiTO Federal State Budgetary Institution «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation, carried out under the European Convention for the Protection of Vertebrate Animals Used for Experiments or Other Scientific Purposes (ETS N 123), Directive of the European Parliament and Council of the European Union 2010/63/EU of 22.09.2010 on the protection of animals used for scientific purposes.

Contribution:
Yurchenko V.V. — concept and design of the study, work with animals, preparation of preparations for cytogenetic analysis, statistical analysis, analysis of literature, writing a text;
Akhaltseva L.V. — cytogenetic analysis, search for literature sources, analysis of literature;
Yurtseva N.A. — work with animals, preparation of preparations for cytogenetic analysis, search for literature sources;
Konyashkina M.A. — work with animals, preparation of preparations for cytogenetic analysis, cytogenetic analysis, search for literature sources;
Lebedev A.S. — verification of Ponceau 4R substances for identity. All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version.

Conflict of interest. The authors declare no conflict of interest.

Acknowledgment. The work was carried out within the framework of the state task «Complex system for assessing the genotoxicity of food additives» Centre for Strategic Planning of FMBA of Russia.

Received: August 9, 2023 / Accepted: November 15, 2023 / Published: December 8, 2023