The effect of a mixture of food additives on the chemical composition of the brain and cognitive functions of experimental animals

Introduction. In conditions of high chemical load and the increasing prevalence of neurodegenerative diseases, it seems relevant to study the effect of the most common food additives on cognitive functions.

The aim is to study the effect of increased doses of sodium benzoate, potassium sorbate, and ascorbic acid on the cognitive functions in white laboratory rats in a rectangular maze, taking into account the ability of the animal’s body to recover after exposure to this mixture.

Materials and methods. The study of changes in the chemical composition of the brain under the influence of a mixture of food additives at different dose levels by capillary electrophoresis was carried out in a series of experiments using laboratory mice; rats were used to assess changes in cognitive function under the influence of a mixture of food additives. For statistical processing of the obtained data, the IBM SPSS Statistics 21 package was used. Comparison of groups was carried out using one-way variance analysis.

Results. Dietary supplements have been shown to affect the chemical composition of the brain in mice by altering the concentrations of ascorbic (AA), sorbic (SA) and benzoic acids (BA). An increase in the concentration of AA in the brain was observed in all groups receiving additives, especially on day 30 of the experiment. The maximum concentration of SA was recorded at different time points of the experiment (day 5, day 12) depending on the dose of preservatives. An increased content of BA was shown on day 12 of the experiment. During the initial training phase, the supplement mixture improved the rats’ cognitive performance, reducing maze time in both sexes. On day 8, differences in the speed of completing the maze were noted between females and males (p=0.026). After the recovery stage, a deterioration in performance was recorded in both sexes, but more pronounced in females. On day 13, the best results in completing the maze were recorded in female rats.

Limitations. The study was conducted once on two types of laboratory rodents. The results obtained on rats and mice cannot always be directly transferred to humans.

Conclusions. Changes in the chemical composition of the brain in animals were observed by the 30^th day of the experiment, which indirectly correlates with some changes in cognitive functions. A neuroprotective effect of ascorbic acid is assumed, but further studies are needed to clarify the potential mechanism of the processes.

Compliance with ethical standards. Date of the meeting of the bioethics commission of the Ufa Research Institute of Occupational Medicine and Human Ecology 02. 08.2024 No. 01-02. The studies were carried out in accordance with the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes (ETS N 123), Directive of the European Parliament and of the Council of the European Union 2010/63/EU of 09.22.2010 on the protection of animals used for scientific purposes.

Contribution:
Smolyankin D.A., Ryabova Yu.V. ‒ study concept and design, collection and processing of materials, statistical processing of data, writing, editing;
Khusnutdinova N.Yu., Repina E.F., Valova Ya.V., Yakupova T.G., Khmel A.O. ‒ collection and processing of materials, editing;
Kurilov M.V. ‒ collection and processing of materials, statistical processing of data, editing;
Karimov D.O. ‒ study concept and design, statistical processing of data, writing, editing;
Karimov D.D. ‒ collection and processing of materials, statistical processing of data;
Akhmadeev A.R. ‒ collection and processing of material.
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.

Funding. The work was carried out using funds from a subsidy for the implementation of a state assignment within the framework of the industry research program of Rospotrebnadzor for 2021–2025. “Scientific substantiation of the national system for ensuring sanitary and epidemiological well-being, managing health risks and improving the quality of life of the population of Russia”, paragraph 6.1.8 “Scientific substantiation of approaches to assessing the toxic effect of xenobiotics based on cellular technologies and model objects”, registration number: 121062100058-8.

Received: February 2, 2025 / Accepted: June 26, 2025 / Published: March 13, 2026

1. Dehghan P., Mohammadi A., Mohammadzadeh-Aghdash H., Ezzati Nazhad Dolatabadi J. Pharmacokinetic and toxicological aspects of potassium sorbate food additive and its constituents. Trends Food Sci. Technol. 2018; 80: 123–30. https://doi.org/10.1016/j.tifs.2018.07.012

2. Fathi F., Ezzati Nazhad Dolatanbadi J., Rashidi M.R., Omidi Y. Kinetic studies of bovine serum albumin interaction with PG and TBHQ using surface plasmon resonance. Int. J. Biol. Macromol. 2016; 91: 1045–50. https://doi.org/10.1016/j.ijbiomac.2016.06.054

3. Yang W., Wu Z., Huang Z.Y., Miao X. Preservation of orange juice using propolis. J. Food Sci. Technol. 2017; 54(11): 3375–83. https://doi.org/10.1007/s13197-017-2754-x

4. Helal E.G., El-Sayed R.A., El-Gamal M.S. Assessment of the physiological changes induced by sodium nitrite, annatto or mono sodium glutamate in male albino rats. Egypt. J. Hosp. Med. 2017; 67(1): 330–5.

5. Piper J.D., Piper P.W. Benzoate and sorbate salts: a systematic review of the potential hazards of these invaluable preservatives and the expanding spectrum of clinical uses for sodium benzoate. Compr. Rev Food Sci. Food Saf. 2017; 16(5): 868–80. https://doi.org/10.1111/1541-4337.12284

6. Kehinde O.S., Christianah O.I., Oyetunji O.A. Ascorbic acid and sodium benzoate synergistically aggravates testicular dysfunction in adult Wistar rats. Int. J. Physiol. Pathophysiol. Pharmacol. 2018; 10(1): 39–46.

7. Mahmoud G.S., Sayed S.A., Abdelmawla S.N., Amer M.A. Positive effects of systemic sodium benzoate and olanzapine treatment on activities of daily life, spatial learning and working memory in ketamine-induced rat model of schizophrenia. Int. J. Physiol. Pathophysiol. Pharmacol. 2019; 11(2): 21–30.

8. Beezhold B.L., Johnston C.S., Nochta K.A. Sodium benzoate-rich beverage consumption is associated with increased reporting of ADHD symptoms in college students: a pilot investigation. J. Atten. Disord. 2014; 18(3): 236–41. https://doi.org/10.1177/1087054712443156

9. Helal E.G., Abdelaziz M.A., EL-Shenawe N.S. Adverse effects of two kinds of food additive mixtures (Sodium benzoate+Monosodium glutamate, Monosodium glutamate+Chlorophyllin and Sodium benzoate+Chlorophyllin) on some physiological parameters in male albino rats. Egypt. J. Hosp. Med. 2019; 75(4): 2736–44.

10. Gaur H., Purushothaman S., Pullaguri N., Bhargava Y., Bhargava A. Sodium benzoate induced developmental defects, oxidative stress and anxiety-like behaviour in zebrafish larva. Biochem. Biophys. Res. Commun. 2018; 502(3): 364–9. https://doi.org/10.1016/j.bbrc.2018.05.171

11. Khoshnoud M.J., Siavashpour A., Bakhshizadeh M., Rashedinia M. Effects of sodium benzoate, a commonly used food preservative, on learning, memory, and oxidative stress in brain of mice. J. Biochem. Mol. Toxicol. 2018; 32(2): e22022. https://doi.org/10.1002/jbt.22022

12. Lin C.H., Chen P.K., Chang Y.C., Chuo L.J., Chen Y.S., Tsai G.E., et al. Benzoate, a D-amino acid oxidase inhibitor, for the treatment of early-phase Alzheimer disease: a randomized, double-blind, placebo-controlled trial. Biol. Psychiatry. 2014; 75(9): 678–85. https://doi.org/10.1016/j.biopsych.2013.08.010

13. Hoang Y.T., Vu A.T. Sodium benzoate and potassium sorbate in processed meat products collected in Ho Chi Minh City, Vietnam. Int. J. Adv. Sci. Eng. Inf. Technol. 2016; 6(4): 477–82.

14. Noorafshan A., Erfanizadeh M., Karbalay-Doust S. Sodium benzoate, a food preservative, induces anxiety and motor impairment in rats. Neurosciences (Riyadh). 2014; 19(1): 24–8.

15. Hasson A. The effect of induced oxidative stress by short and long-terms exposure to potassium sorbate (E202) as a food additive on the female reproductive system of Wistar rats. Int. J. Pharm. Res. 2020; 12: 1407–22. https://doi.org/10.31838/ijpr/2020.SP2.167

16. Hussain S., Riaz A., Ali M., Ullah N., Hussain N. Quality assessment of sweet cherry (Prunus avium) juice treated with different chemical preservatives. J. Food Process Technol. 2019; 10(786): 2.

17. Taghavi F., Habibi-Rezaei M., Bohlooli M., Farhadi M., Goodarzi M., Movaghati S., et al. Antiamyloidogenic effects of ellagic acid on human serum albumin fibril formation induced by potassium sorbate and glucose. J. Mol. Recognit. 2016; 29(12): 611–8. https://doi.org/10.1002/jmr.2560

18. Gulcin İ. Antioxidants and antioxidant methods: an updated overview. Arch. Toxicol. 2020; 94(3): 651–715. https://doi.org/10.1007/s00204-020-02689-3

19. Rahaman M.M., Hossain R., Herrera-Bravo J., Islam M.T., Atolani O., Adeyemi O.S., et al. Natural antioxidants from some fruits, seeds, foods, natural products, and associated health benefits: An update. Food Sci. Nutr. 2023; 11(4): 1657–70. https://doi.org/10.1002/fsn3.3217

20. Martemucci G., Portincasa P., Di Ciaula A., Mariano M., Centonze V., D’Alessandro A.G. Oxidative stress, aging, antioxidant supplementation and their impact on human health: An overview. Mech. Ageing Dev. 2022; 206: 111707. https://doi.org/10.1016/j.mad.2022.111707

21. Bjelakovic G., Nikolova D., Gluud C. Antioxidant supplements and mortality. Curr. Opin. Clin. Nutr. Metab. Care. 2014; 17(1): 40–4. https://doi.org/10.1097/MCO.0000000000000009

22. Al Joudi F.S. Adverse effects of excessive antioxidant supplements and their underlying mechanisms. J. Aging Res. Clin. Pract. 2013; 2(4): 339–45.

23. Harvie M. Nutritional supplements and cancer: potential benefits and proven harms. Am. Soc. Clin. Oncol. Educ. Book. 2014: e478–86. https://doi.org/10.14694/EdBook_AM.2014.34.e478

24. Shahid M., Alwan N.A., Al-Masoudi E.A. A study of toxic effect of sodium benzoate, vit. C alone and their combination on reproductive functions of adult male rabbits. Basrah J. Vet. Res. 2018; 17(3): 533–43.

25. Onaolapo A.Y., Onaolapo O.J. Neuroprotection induced by ascorbic acid. In: Natural Molecules in Neuroprotection and Neurotoxicity. Academic Press: 2024: 1263–88.

26. Avdeeva O.I., Karachinskaya I.V., Abrashova T.V., Dray R.V., Makarova M.N., Makarov V.G., et al. Toxicological study of ascorbic acid high doses. Profilakticheskaya i klinicheskaya meditsina. 2011; (1): 40–4. https://elibrary.ru/rymgtv (in Russian)

27. Doseděl M., Jirkovský E., Macáková K., Krčmová L.K., Javorská L., Pourová J., et al. Vitamin C-sources, physiological role, kinetics, deficiency, use, toxicity, and determination. Nutrients. 2021; 13(2): 615. https://doi.org/10.3390/nu13020615

28. Gęgotek A., Skrzydlewska E. Ascorbic acid as antioxidant. Vitam. Horm. 2023; 121: 247–70. https://doi.org/10.1016/bs.vh.2022.10.008

29. Helal E.G., Mustafa R.A., Mohamed A., El-Gamal M.S. Adverse effects of two kinds of food additive mixtures (flavor enhancer, food preservative or food coloring agent) on physiological parameters in young male albino rats. Egypt. J. Hosp. Med. 2017; 67(1): 344–51. https://doi.org/10.12816/0036646

30. Radwan E.H., Elghazaly M.M., Hussein H.K., Abdel Aziz K.K., Barakat A.I. Adverse effect of mixture of food additives on some biochemical parameters in male albino rats. J. Adv. Biol. 2020; 13: 1–13. https://doi.org/10.24297/jab.v13i.8555

31. Nzeh B.C., Chiegboka N.A., Nwanyanwu C.E., Asiwe E.S. Inhibitory and interactive effects of mixtures of chemical preservatives against food spoilage bacteria. Eur. J. Biomed. 2019; 6(11): 264–74.

32. Oladapo A., Akinyosoye F.A., Abiodun O.A. The inhibitory effect of different chemical food preservatives on the growth of selected food borne pathogenic bacteria. Afr. J. Microbiol. Res. 2014; 8(14): 1510–5. https://doi.org/10.5897/AJMR2013.6370

33. Heshmati A., Ghadimi S., Mousavi Khaneghah A., Barba F.J., Lorenzo J.M., Nazemi F., et al. Risk assessment of benzene in food samples of Iran’s market. Food Chem. Toxicol. 2018; 114: 278–84. https://doi.org/10.1016/j.fct.2018.02.043

34. Khodaei F., Kholghipour H., Hosseinzadeh M., Rashedinia M. Effect of sodium benzoate on liver and kidney lipid peroxidation and antioxidant enzymes in mice. J. Rep. Pharm. Sci. 2019; 8(2): 217. https://doi.org/10.4103/jrptps.JRPTPS_68_18

35. Hörnberg H., Pohl T. Neuroligins in neurodevelopmental conditions: How mouse models of de novo mutations can help us link synaptic function to social behavior. Neuronal Signal. 2022; 6(2): NS20210030. https://doi.org/10.1042/NS20210030

36. DeGrazia, D., Beauchamp T.L. Beyond the 3 Rs to a more comprehensive framework of principles for animal research ethics. ILAR J. 2021; 60(3): 308–17. https://doi.org/10.1093/ilar/ilz011

37. Barrington W.T., Wulfridge P., Wells A.E., Rojas C.M., Howe S.Y.F., Perry A., et al. Improving metabolic health through precision dietetics in mice. Genetics. 2018; 208(1): 399–417. https://doi.org/10.1534/genetics.117.300536

38. Huang W., Lin Z., Sun A., Deng J.M., Manyande A., Xiang H., et al. The role of gut microbiota in diabetic peripheral neuropathy rats with cognitive dysfunction. Front. Microbiol. 2023; 14: 1156591. https://doi.org/10.3389/fmicb.2023.1156591

39. Bick S.K., Eskandar E.N. Neuromodulation for restoring memory. Neurosurg. Focus. 2016; 40(5): E5. https://doi.org/10.3171/2016.3.FOCUS162

40. Matsuura A., Fujita Y., Iyo M., Hashimoto K. Effects of sodium benzoate on pre-pulse inhibition deficits and hyperlocomotion in mice after administration of phencyclidine. Acta Neuropsychiatr. 2015; 27(3): 159–67. https://doi.org/10.1017/neu.2015.1

41. Helal E., Barayan A., Abdelaziz M., El-Shnawe N. Adverse effects of mono sodium glutamate, sodium benzoate and chlorophyllins on some physiological parameters in male albino rats. Egypt. J. Hosp. Med. 2019; 74(8): 1857–64. https://doi.org/10.21608/ejhm.2019.28865

42. Lane H.Y., Lin C.H., Green M.F., Hellemann G., Huang C.C., Chen P.W., et al. Add-on treatment of benzoate for schizophrenia: a randomized, double-blind, placebo-controlled trial of D-amino acid oxidase inhibitor. JAMA Psychiatry. 2013; 70(12): 1267–75. https://doi.org/10.1001/jamapsychiatry.2013.2159

43. Hovatta I., Tennant R.S., Helton R., Marr R.A., Singer O., Redwine J.M., et al. Glyoxalase 1 and glutathione reductase 1 regulate anxiety in mice. Nature. 2005; 438(7068): 662–6. https://doi.org/10.1038/nature04250

44. Sepehri H., Hojati A., Safari R. Effect of bitter melon on spatial memory of rats receiving a high-fat diet. J. Exp. Pharmacol. 2019; 11: 115–9. https://doi.org/10.2147/JEP.S231260

45. Zhang X., Dong F., Ren J., Driscoll M.J., Culver B. High dietary fat induces NADPH oxidase-associated oxidative stress and inflammation in rat cerebral cortex. Exp. Neurol. 2005; 191(2): 318–25. https://doi.org/10.1016/j.expneurol.2004.10.011

46. Souza C.G., Moreira J.D., Siqueira I.R., Pereira A.G., Rieger D.K., Souza D.O., et al. Highly palatable diet consumption increases protein oxidation in rat frontal cortex and anxiety-like behavior. Life Sci. 2007; 81(3): 198–203. https://doi.org/10.1016/j.lfs.2007.05.001

47. El-Nouby K.A., Hamouda H.E., Abd El Azeem M.A., El-Ebiary A.A. Food additives and Hymenolepis nana infection: an experimental study. J. Egypt. Soc. Parasitol. 2009; 39(3): 1015–32.

48. DiGirolamo A.M., Ramirez-Zea M., Wang M., Flores-Ayala R., Martorell R., Neufeld L.M., et al. Randomized trial of the effect of zinc supplementation on the mental health of school-age children in Guatemala. Am. J. Clin. Nutr. 2010; 92(5): 1241–50. https://doi.org/10.3945/ajcn.2010.29686

49. Markel A.L., Achkasov A.F., Alekhina T.A., Prokudina O.I., Ryazanova M.A., Ukolova T.N., et al. Effects of the alpha- and gamma-polymorphs of glycine on the behavior of catalepsy prone rats. Pharmacol. Biochem. Behav. 2011; 98(2): 234–40. https://doi.org/10.1016/j.pbb.2010.12.025

50. Badenhorst C.P., Erasmus E., van der Sluis R., Nortje C., van Dijk A.A. A new perspective on the importance of glycine conjugation in the metabolism of aromatic acids. Drug Metab. Rev. 2014; 46(3): 343–61. https://doi.org/10.3109/03602532.2014.908903

51. Quadros L., Brandao I., Longhi R. Ascorbic acid and performance: A review. Vitam. Miner. 2016; 5(1): 136–40.

52. Li S., Tan H.Y., Wang N., Zhang Z.J., Lao L., Wong C.W., et al. The role of oxidative stress and antioxidants in liver diseases. Int. J. Mol. Sci. 2015; 16(11): 26087–124. https://doi.org/10.3390/ijms161125942

53. Jana A., Modi K.K., Roy A., Anderson J.A., van Breemen R.B., Pahan K. Up-regulation of neurotrophic factors by cinnamon and its metabolite sodium benzoate: therapeutic implications for neurodegenerative disorders. J. Neuroimmune Pharmacol. 2013; 8(3): 739–55. https://doi.org/10.1007/s11481-013-9447-7

54. Brahmachari S., Jana A., Pahan K. Sodium benzoate, a metabolite of cinnamon and a food additive, reduces microglial and astroglial inflammatory responses. J. Immunol. 2009; 183(9): 5917–27. https://doi.org/10.4049/jimmunol.0803336

55. Smolyankin D.A., Khusnutdinova N.Yu., Kurilov M.V., Kutlina T.G., Timasheva G.V. Study of some physiological tests in white inspired mice during the introduction of various doses of a preservation mixture. Meditsina truda i ekologiya cheloveka. 2019; (2): 80–3. https://elibrary.ru/xctujj (in Russian)