medlitГигиена и санитарияHygiene and Sanitation0016-99002412-0650Federal Scientific Center of Hygiene named after F.F. Erisman10.47470/0016-9900-2022-101-7-798-801medlit-2377Research ArticleГИГИЕНА ПИТАНИЯFOOD HYGIENEОценка генотоксичности пищевого красителя тартразина в микроядерном тесте in vivoEvaluation of the genotoxicity of the food dye tartrazine in a micronucleus test in vivohttps://orcid.org/0000-0002-3619-3858АхальцеваЛюдмила ВячеславовнаAkhaltsevaLyudmila V.

Канд. биол. наук, ст. науч. сотр. отд. профилактической токсикологии и медико-биологических исследований ФГБУ «Центр стратегического планирования и управления медико-биологическими рисками здоровью» ФМБА России, 119121, Москва.

e-mail: LAhalceva@cspmz.ru

MD, PhD, Senior 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: LAhalceva@cspmz.ru

lahalceva@cspmz.ruhttps://orcid.org/0000-0003-4377-245XЮрченкоВ. В.YurchenkoValentina V.noemail@neicon.ruhttps://orcid.org/0000-0001-5031-2916ЮрцеваН. А.YurtsevaNadezda A.noemail@neicon.ruhttps://orcid.org/0000-0002-8319-1329КоняшкинаМ. А.KonyashkinaMariya A.noemail@neicon.ruФГБУ «Центр стратегического планирования и управления медико-биологическими рисками здоровью» Федерального медико-биологического агентстваРоссияCentre for Strategic Planning of FMBA of RussiaRussian Federation2022100820221017798801Copyright © Ахальцева Л.В., Юрченко В.В., Юрцева Н.А., Коняшкина М.А., 20222022Ахальцева Л.В., Юрченко В.В., Юрцева Н.А., Коняшкина М.А.Akhaltseva L.V., Yurchenko V.V., Yurtseva N.A., Konyashkina M.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/2377Введение

Введение. Пищевой азокраситель тартразин (Е102) широко применяется при производстве продуктов питания, фармакологических и косметических средств. Несмотря на разрешение к применению, интерес к всесторонней оценке влияния пищевых красителей, особенно синтетических, на здоровье не ослабевает. Анализ данных литературы по оценке генотоксичности тартразина при исследованиях in vivo выявил некоторые противоречия результатов, что показало целесообразность испытания приобретённого в розничной продаже пищевого красителя в одном из рекомендованных тестов.

Материалы и методы

Материалы и методы. Генотоксичность тартразина (производство Индия, чистота 88,37%) изучена в микроядерном тесте на клетках костного мозга самцов мышей гибридов F1(CBAxC57Bl6/j). Исследуемое вещество двукратно вводили в желудок мышей в интервале доз 250–2000 мг/кг. Частоту полихроматофильных эритроцитов (ПХЭ) с микроядрами (МЯ) оценивали по результатам анализа 4000 ПХЭ. Долю ПХЭ среди всех эритроцитов определяли при анализе 1000 клеток на каждое животное.

Результаты

Результаты. Не выявлено повышения частоты полихроматофильных эритроцитов с МЯ над параллельным отрицательным контролем при двукратном энтеральном введении тартразина во всех изученных дозах. Доля ПХЭ среди всех эритроцитов не изменялась.

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

Заключение

Заключение. Образец пищевого красителя тартразина (Е102) в диапазоне доз 250–2000 мг/кг не проявил цитогенетической активности в микроядерном тесте in vivo на клетках костного мозга мышей при двукратном введении.

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: Ахальцева Л.В. — цитомный анализ, поиск источников литературы, анализ данных литературы, написание текста; Юрченко В.В. — концепция и дизайн исследования, работа с животными, приготовление препаратов для цитомного анализа, статистический анализ, анализ данных литературы, написание текста; Юрцева Н.А. — работа с животными, приготовление препаратов для цитомного анализа, цитомный анализ, поиск источников литературы; Коняшкина М.А. — работа с животными, приготовление препаратов для цитомного анализа, поиск источников литературы, анализ данных 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: 21.02.2022 / Принята к печати: 08.06.2022 / Опубликована: 31.07.2022

Introduction

Introduction. Food azo dye Tartrazine (E102) is widely used in the production of food, pharmacological and cosmetic products. Despite the approval for use, interest in a comprehensive assessment of the impact of food colours, especially synthetic ones, on health continues unabated. The analysis of literature evaluating the genotoxicity of Tartrazine in vivo studies revealed some inconsistencies in the results, that showed the possibility to test a retail food colouring in one of recommended tests.

Materials and methods

Materials and methods. The Tartrazine genotoxicity (produced in India, purity 88.37%) was studied in the micronucleus test on male mouse bone marrow cells (hybrids F1 CBA x C57Bl6/j). The test substance was double enteral administrated in the dose range 250–2000 mg/kg. The frequency of polychromatophilic erythrocytes (PCEs) with micronuclei (MNs) was estimated by the analysis of 4000 PCEs. The proportion of PCE among all erythrocytes was determined by analyzing 1000 cells per animal.

Results

Results. There was no increase in the frequency of micronucleated polychromatophilic erythrocytes over a concurrent negative control with double enteral administration of Tartrazine in all doses studied. The proportion of PCEs among all erythrocytes did not change.

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. The sample of the food dye Tartrazine (E 102) in the dose range of 250-2000 mg/kg did not show cytogenetic activity in the in vivo micronucleus test on mouse bone marrow cells after a double enteral administration.

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 No. 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: Akhaltseva L.V. — cytome analysis, search for literature sources, analysis of literature, writing a text; Yurchenko V.V. — concept and design of the study, work with animals, preparation of preparations for cytome analysis, statistical analysis, analysis of literature, writing a text; Yurtseva N.A. — work with animals, preparation of preparations for cytome analysis, cytome analysis, search for literature sources; Konyashkina M.A. — work with animals, preparation of preparations for cytome analysis, search for literature sources, analysis of literature. All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version. 

Conflict of interest

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

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.

Received

Received: February 21, 2022 / Accepted: June 08, 2022 / Published: July 31, 2022

генотоксичность пищевых синтетических азокрасителейтартразинмикроядерный тест in vivoполихроматофильные эритроцитыкостный мозгмышиgenotoxicity of food synthetic azo dyesTartrazinein vivo micronucleus testpolychromatophilic erythrocytesbone marrowmiceReferencesKobylewski S., Jacobson M.F. Toxicology of food dyes.Int. J. Occup. Environ. Health. 2012; 18(3): 220-46. https://doi.org/10.1179/1077352512z.00000000034Kobylewski S., Jacobson M.F. Toxicology of food dyes. Int. J. Occup. Environ. Health. 2012; 18(3): 220–46. https://doi.org/10.1179/1077352512z.00000000034ГИОРД. Мировой рынок пищевых красителей: интенсивный рост натурального сегмента. Доступно: https://www.giord.ru/news/novosti_otrasli/mirovoy_rynok_pishchevykh_krasiteley_intensivnyy_rost_naturalnogo_segmenta/GIORD. The world market of food dyes: intensive growth of the natural segment. Available at: https://www.giord.ru/news/novosti_otrasli/mirovoy_rynok_pishchevykh_krasiteley_intensivnyy_rost_naturalnogo_segmenta/ (in Russian)European Food Safety Authority. Scientific opinion on the re-evaluation of tartrazine (E 102) as a food additive. EFSA J. 2009; 7(11): 1331-82. https://doi.org/10.2903/j.efsa.2009.1331European Food Safety Authority. Scientific opinion on the re-evaluation of tartrazine (E 102) as a food additive. EFSA J. 2009; 7(11): 1331–82. https://doi.org/10.2903/j.efsa.2009.1331WHO. Food Additives Series, No. 73. Safety evaluation of certain food additives: prepared by the Eighty-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Geneva; 2017.WHO. Food Additives Series, No. 73. Safety evaluation of certain food additives: prepared by the Eighty-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Geneva; 2017.EMA/CHMP/ICH/126642/2008. ICH guideline S2 (R1) on genotoxicity testing and data interpretation for pharmaceuticals intended for human use. European Medicines Agency; 2013. Available at: https://www.ema.europa.eu/en/documents/scientific-guideline/ich-guideline-s2-r1-genotoxicity-testing-data-interpretation-pharmaceuticals-intended-human-use-step_en.pdfEMA/CHMP/ICH/126642/2008. ICH guideline S2 (R1) on genotoxicity testing and data interpretation for pharmaceuticals intended for human use. European Medicines Agency; 2013. Available at: https://www.ema.europa.eu/en/documents/scientific-guideline/ich-guideline-s2-r1-genotoxicity-testing-data-interpretation-pharmaceuticals-intended-human-use-step_en.pdfCommittee on Mutagenicity of Chemicals in Food, Consumer Products, and the Environment.Interim guidance on a strategy for genotoxicity testing and mutagenic hazard assessment of impurities in chemical substances; 2012. Available at: https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/1043249/strategy-for-genotoxicity-testing-of-chemicals-guidance.pdfCommittee on Mutagenicity of Chemicals in Food, Consumer Products, and the Environment. Interim guidance on a strategy for genotoxicity testing and mutagenic hazard assessment of impurities in chemical substances; 2012. Available at: https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/1043249/strategy-for-genotoxicity-testing-of-chemicals-guidance.pdfЮрченко В.В., Ингель Ф.И., Ахальцева Л.В., Коняшкина М.А., Юрцева Н.А., Никитина Т.А. и др. Генетическая безопасность синтетических пищевых красителей. Обзор литературы. Экологическая генетика. 2021; 19(4): 323-41. https://doi.org/10.17816/ecogen79399Yurchenko V.V., Ingel’ F.I., Akhal’tseva L.V., Konyashkina M.A., Yurtseva N.A., Nikitina T.A., et al. Genotoxic safety of synthetic food colours. Review. Ekologicheskaya genetika. 2021; 19(4): 323–41. https://doi.org/10.17816/ecogen79399 (in Russian)Commission Regulation (EU) No. 231/2012 laying down specifications for food additives listed in Annexes II and III to Regulation (EC) No. 1333/2008 of the European Parliament and of the CouncilCommission Regulation (EU) No. 231/2012 laying down specifications for food additives listed in Annexes II and III to Regulation (EC) No. 1333/2008 of the European Parliament and of the Council.Руководство Р 1.2.3156-13. Оценка токсичности и опасности химических веществ и их смесей для здоровья человека. М.; 2014Guide Р 1.2.3156-13. Assessment of toxicity and hazards of chemicals and their mixtures for human health. Moscow; 2014. (in Russian)OECD. Test No. 474: Mammalian Erythrocyte Micronucleus Test. OECD Guidelines for the Testing of Chemicals, Section 4. Paris: OECD Publishing; 2016. https://doi.org/10.1787/9789264264762-enOECD. Test No. 474: Mammalian Erythrocyte Micronucleus Test. OECD Guidelines for the Testing of Chemicals, Section 4. Paris: OECD Publishing; 2016. https://doi.org/10.1787/9789264264762-enKornbrust D., Barfknecht T. Testing of 24 food, drug, cosmetic, and fabric dyes in the in vitro and the in vivo/in vitro rat hepatocyte primary culture/DNA repair assay. Environ. Mutagen. 1985; 7(1): 101-20. https://doi.org/10.1002/em.2860070106Kornbrust D., Barfknecht T. Testing of 24 food, drug, cosmetic, and fabric dyes in the in vitro and the in vivo/in vitro rat hepatocyte primary culture/DNA repair assay. Environ. Mutagen. 1985; 7(1): 101–20. https://doi.org/10.1002/em.2860070106Bastaki M., Farrell T., Bhusari S., Pant K., Kulkarni R. Lack of genotoxicity in vivo for food color additive Tartrazine. Food Chem. Toxicol. 2017; 105: 278-84. https://doi.org/10.1016/j.fct.2017.04.034Bastaki M., Farrell T., Bhusari S., Pant K., Kulkarni R. Lack of genotoxicity in vivo for food color additive Tartrazine. Food Chem. Toxicol. 2017; 105: 278–84. https://doi.org/10.1016/j.fct.2017.04.034Sasaki Y.F., Kawaguchi S., Kamaya A., Ohshita M., Kabasawa K., Iwama K., et al. The comet assay with 8 mouse organs: results with 39 currently used food additives. Mutat. Res. 2002; 519(1-2): 103-19. https://doi.org/10.1016/s1383-5718(02)00128-6Sasaki Y.F., Kawaguchi S., Kamaya A., Ohshita M., Kabasawa K., Iwama K., et al. The comet assay with 8 mouse organs: results with 39 currently used food additives. Mutat. Res. 2002; 519(1–2): 103–19. https://doi.org/10.1016/s1383-5718(02)00128-6Abo-El-Sooud K., Hashem M.M., Badr Y.A., Eleiwa M.M.E., Gab-Allaha A.Q., Abd-Elhakim Y.M., et al. Assessment of hepato-renal damage and genotoxicity induced by long-term exposure to five permitted food additives in rats. Environ. Sci. Pollut. Res.Int. 2018; 25(26): 26341-50. https://doi.org/10.1007/s11356-018-2665-zAbo-El-Sooud K., Hashem M.M., Badr Y.A., Eleiwa M.M.E., Gab-Allaha A.Q., Abd-Elhakim Y.M., et al. Assessment of hepato-renal damage and genotoxicity induced by long-term exposure to five permitted food additives in rats. Environ. Sci. Pollut. Res. Int. 2018; 25(26): 26341–50. https://doi.org/10.1007/s11356-018-2665-zHassan G.M. Effects of some synthetic coloring additives on DNA damage and chromosomal aberrations of rats. Arab. J. Biotech. 2010; 13(1): 13-24.Hassan G.M. Effects of some synthetic coloring additives on DNA damage and chromosomal aberrations of rats. Arab. J. Biotech. 2010; 13(1): 13–24.El-Desoky G.E., Wabaidur S.M., AlOthman Z.A., Habila M.A. Regulatory role of nano-curcumin against tartrazine-induced oxidative stress, apoptosis-related genes expression, and genotoxicity in rats. Molecules. 2020; 25(24): 5801. https://doi.org/10.3390/molecules25245801El-Desoky G.E., Wabaidur S.M., AlOthman Z.A., Habila M.A. Regulatory role of nano-curcumin against tartrazine-induced oxidative stress, apoptosis-related genes expression, and genotoxicity in rats. Molecules. 2020; 25(24): 5801. https://doi.org/10.3390/molecules25245801Khayyat L., Essawy A., Sorour J., Soffar A. Tartrazine induced structural and functional aberrations and genotoxic effects in vivo. Peer J. 2017; 5: e3041. https://doi.org/10.7717/peerj.3041Khayyat L., Essawy A., Sorour J., Soffar A. Tartrazine induced structural and functional aberrations and genotoxic effects in vivo. Peer J. 2017; 5: e3041. https://doi.org/10.7717/peerj.3041Poul M., Jarry G., Elhkim M.O., Poul J.M. Lack of genotoxic effect of food dyes amaranth, sunset yellow and tartrazine and their metabolites in the gut micro assay in mice. Food Chem. Toxicol. 2009; 47(2): 443-8. https://doi.org/10.1016/j.fct.2008.11.034Poul M., Jarry G., Elhkim M.O., Poul J.M. Lack of genotoxic effect of food dyes amaranth, sunset yellow and tartrazine and their metabolites in the gut micro assay in mice. Food Chem. Toxicol. 2009; 47(2): 443–8. https://doi.org/10.1016/j.fct.2008.11.034Дурнев А.Д., Орещенко А.В., Кулакова А.В., Берестень Н.Ф. Анализ цитологической активности пищевых красителей. Вопросы медицинской химии. 1995; (5): 50-3Durnev A.D., Oreshchenko A.V., Kulakova A.V., Beresten N.F. Analysis of cytogenetic activity of food dyes. Voprosy meditsinskoy khimii. 1995; (5): 50–3. (in Russian)Das A., Mukherjee A. Genotoxicity testing of the food colours amaranth and tartrazine.Int. J. Hum. Genet. 2004; 4(4): 277-80. https://doi.org/10.1080/09723757.2004.11885906Das A., Mukherjee A. Genotoxicity testing of the food colours amaranth and tartrazine. Int. J. Hum. Genet. 2004; 4(4): 277–80. https://doi.org/10.1080/09723757.2004.11885906Giri A.K., Das S.K., Talukder G., Sharma A. Sister chromatid exchange and chromosome aberrations induced by curcumin and tartrazine on mammalian cells in vivo. Cytobios. 1990; 62(249): 111-7.Giri A.K., Das S.K., Talukder G., Sharma A. Sister chromatid exchange and chromosome aberrations induced by curcumin and tartrazine on mammalian cells in vivo. Cytobios. 1990; 62(249): 111–7.Farag I.M., Abdel Aziz K.B., El-Nahass E., Zaher M., Hegazy M.A., Roshdy H.M. Cytogenetic studies on the effects of Tartrazine, beta-carotene and a mixture of both dyes on pregnant mice and their embryos. Bull. Natl Res. Cent. (Egypt). 2001; 26(1): 93-109.Farag I.M., Abdel Aziz K.B., El-Nahass E., Zaher M., Hegazy M.A., Roshdy H.M. Cytogenetic studies on the effects of Tartrazine, beta-carotene and a mixture of both dyes on pregnant mice and their embryos. Bull. Natl Res. Cent. (Egypt). 2001; 26(1): 93–109.OECD. Test No. 475: Mammalian Bone Marrow Chromosomal Aberration Test. OECD Guidelines for the Testing of Chemicals; Section 4. Paris: OECD Publishing; 2016. https://doi.org/10.1787/9789264264786-enOECD. Test No. 475: Mammalian Bone Marrow Chromosomal Aberration Test. OECD Guidelines for the Testing of Chemicals; Section 4. Paris: OECD Publishing; 2016. https://doi.org/10.1787/9789264264786-enOECD. Test No. 489: In Vivo Mammalian Alkaline Comet Assay. OECD Guidelines for the Testing of Chemicals; Section 4. Paris: OECD Publishing; 2016. https://doi.org/10.1787/9789264264885-enOECD. Test No. 489: In Vivo Mammalian Alkaline Comet Assay. OECD Guidelines for the Testing of Chemicals; Section 4. Paris: OECD Publishing; 2016. https://doi.org/10.1787/9789264264885-enBrown J.P. Role of gut bacterial flora in nutrition and health: a review of recent advances in bacteriological techniques, metabolism and factors affecting flora composition. Crit. Rev. Food Sci. Nutr. 1977; 8(3): 229. https://doi.org/10.1080/10408397709527224Brown J.P. Role of gut bacterial flora in nutrition and health: a review of recent advances in bacteriological techniques, metabolism and factors affecting flora composition. Crit. Rev. Food Sci. Nutr. 1977; 8(3): 229. https://doi.org/10.1080/10408397709527224Brown J.P. Reduction of polymeric azo and nitro dyes by intestinal bacteria. Appl. Environ. Microbiol. 1981; 41(5): 1283-6. https://doi.org/10.1128/aem.41.5.1283-1286.1981Brown J.P. Reduction of polymeric azo and nitro dyes by intestinal bacteria. Appl. Environ. Microbiol. 1981; 41(5): 1283–6. https://doi.org/10.1128/aem.41.5.1283-1286.1981

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