medlitГигиена и санитарияHygiene and Sanitation0016-99002412-0650Federal Scientific Center of Hygiene named after F.F. Erisman10.47470/0016-9900-2023-102-6-580-583qhujwymedlit-3197Research ArticleГИГИЕНА ПИТАНИЯFOOD HYGIENEОценка цитогенетической активности пищевого красителя азорубина в микроядерном тесте на мышахEvaluation of the cytogenetic activity of the food dye Azorubine in a micronucleus test in micehttps://orcid.org/0000-0003-4377-245XЮрченкоВалентина ВасильевнаYurchenkoValentina V.

Канд. мед. наук, вед. науч. сотр. отд. профилактической токсикологии и медико-биологических исследований ФГБУ «ЦСП» ФМБА России, 119121, Москва.

e-mail: VYurchenko@cspmz.ru

MD, PhD. Leading Researcher of the Department of Preventive Toxicology and Biomedical Research in the Centre for Strategic Planning of FMBA of Russia, Moscow, 119121, Russian Federation.

e-mail: VYurchenko@cspmz.ru

VYurchenko@cspmz.ruhttps://orcid.org/0000-0002-3619-3858АхальцеваЛюдмила В.AkhaltsevaLyudmila V.noemail@neicon.ruhttps://orcid.org/0000-0002-8319-1329КоняшкинаМария А.KonyashkinaMariya A.noemail@neicon.ruhttps://orcid.org/0000-0001-5031-2916ЮрцеваНадежда А.YurtsevaNadezda A.noemail@neicon.ruФГБУ «Центр стратегического планирования и управления медико-биологическими рисками здоровью» Федерального медико-биологического агентстваCentre for Strategic Planning of FMBA of Russia2023310720231026580583Copyright © Юрченко В.В., Ахальцева Л.В., Коняшкина М.А., Юрцева Н.А., 20232023Юрченко В.В., Ахальцева Л.В., Коняшкина М.А., Юрцева Н.А.Yurchenko V.V., Akhaltseva L.V., Konyashkina M.A., Yurtseva N.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.rjhas.ru/jour/article/view/3197Введение

Введение. Моноазокраситель азорубин Е122 (кармуазин, пищевой красный 3) используют при изготовлении десертов, карамелей, конфет, мармеладов, мороженого, алкогольных и безалкогольных напитков и т. д. Оценка безопасности пищевых добавок включает изучение генотоксического потенциала. При этом для веществ с высокой экспозицией или небольшой, но длительной (в том числе для пищевых добавок) обязательны тесты in vivo.

Материалы и методы

Материалы и методы. Микроядерным методом на клетках костного мозга мышей (самцы, гибриды F1 CBA × C57Bl6/j) изучена генотоксическая активность водного раствора синтетического пищевого азокрасителя азорубина Е122. Исследуемые вещества вводили в желудок мышей в диапазоне доз 250–2000 мг/кг двукратно с интервалом 24 ч, приготовление препаратов осуществляли через 24 ч после последнего введения. Частоту полихроматофильных эритроцитов (ПХЭ) с микроядрами (МЯ) оценивали по результатам анализа 4000 ПХЭ, долю ПХЭ — по результатам анализа 500 эритроцитов на каждое животное.

Результаты

Результаты. Не выявлено статистически значимого повышения частоты ПХЭ с МЯ по отношению к параллельному контролю при двукратном введении азорубина во всех изученных дозах. После воздействия в дозах 1000 и 2000 мг/кг частота ПХЭ с МЯ несколько превышала верхний предел 95%-го ДИ накопленного отрицательного контроля. Эффект статистически значимо зависел от дозы, что не позволяет признать ответ чётко отрицательным.

Ограничения исследования обусловлены методологией теста: проанализированы только цитогенетические нарушения в единственной ткани в условиях двукратного введения изученного образца.

Заключение

Заключение. Анализ частоты ПХЭ с МЯ в костном мозге мышей после двукратного введения азорубина в дозах 250–2000 мг/кг позволил квалифицировать результат эксперимента как неопределённый.

Соблюдение этических стандартов. Исследование одобрено локальным этическим комитетом НИИ ЭДиТО ФГБУ «НМИЦ онкологии им. Н.Н. Блохина» Минздрава России, проведено в соответствии с Европейской конвенцией о защите позвоночных животных, используемых для экспериментов или в иных научных целях (ETS N 123), директивой Европейского парламента и Совета Европейского союза 2010/63/EC от 22.09.2010 г. о защите животных, использующихся для научных целей.

Участие авторов

Участие авторов: Юрченко В.В. — концепция и дизайн исследования, работа с животными, приготовление препаратов для цитогенетического анализа, статистический анализ, анализ данных литературы, написание текста; Ахальцева Л.В. — цитогенетический анализ, поиск источников литературы, анализ данных литературы; Коняшкина М.А. — работа с животными, приготовление препаратов для цитогенетического анализа; цитогенетический анализ, поиск источников литературы; Юрцева Н.А. — работа с животными, приготовление препаратов для цитогенетического анализа, поиск источников литературы. Все соавторы — утверждение окончательного варианта статьи, ответственность за целостность всех частей статьи.

Конфликт интересов

Конфликт интересов. Авторы декларируют отсутствие явных и потенциальных конфликтов интересов в связи с публикацией данной статьи.

Финансирование

Финансирование. Работа выполнена в рамках государственного задания «Комплексная система оценки генотоксичности пищевых добавок» ФГБУ «ЦСП» ФМБА России.

Поступила

Поступила: 06.02.2023 / Принята к печати: 07.06.2023 / Опубликована: 30.07.2023

Introduction

Introduction. Azorubine E122 (Carmoisine, Food Red 3) monoazo dye is used in the manufacture of desserts, caramels, sweets, marmalades, ice cream, alcoholic and non-alcoholic drinks, etc. The safety assessment of food additives includes the study of genotoxic potential. At the same time, for substances with a high or a small but long-term exposure (including food additives), in vivo tests are required.

Materials and methods

Materials and methods. The genotoxic activity of aqueous solutions of synthetic food azo dyes E122 Azorubin was studied by the micronuclear method on bone marrow cells in mice (males, hybrids F1 CBA × C57Bl6/j). The studied substances were injected into the stomach of mice at doses of 250–2000 mg/kg twice with an interval of 24 hours, with the preparation of bone marrow preparations 24 hours after the last administration. The frequency of micronucleated (MN) polychromatophilic erythrocytes (PCEs) was estimated on the base of the results of the analysis of 4000 PСЕ. The proportion of PСE among all red blood cells was determined by analyzing 500 cells per animal.

Results

Results. There was no statistically significant increase in the frequency of PCE with MN over the current control with a double administration of Azorubine in all studied doses. After exposure at doses of 1000 and 2000 mg/kg, the incidence of MN PCEs slightly exceeded the upper limit of the 95% CI of the accumulated negative control and the effect was dose-dependent and statistically significant, which does not allow recognizing the answer as clearly negative.

Limitations of the study are due to the methodology of the test: only cytogenetic disorders in a single tissue were analyzed under conditions of double enteral administration of the studied sample.

Conclusion

Conclusion. An analysis of the frequency of MN PCEs in the bone marrow of mice after a double injection of Azorubine at doses of 250–2000 mg/kg made it possible to qualify the result of the experiment as uncertain.

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

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; Konyashkina M.A. — work with animals, preparation of preparations for cytogenetic analysis, cytogenetic analysis, search for literature sources; Yurtseva N.A. — work with animals, preparation of preparations for cytogenetic analysis, search for literature sources. All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version.

Acknowledgment

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.

Conflict of interest

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

Received

Received: February 6, 2023 / Accepted: June 7, 2023 / Published: July 30, 2023

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The authors declare that there are no conflicts of interest present.