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GIDA KATKI MADDESİ OLARAK KULLANILAN SİTRİK ASİT, ASKORBİK ASİT VE SODYUM SİTRATIN İNSAN LENFOSİT HÜCRELERİNDE GENOTOKSİSİTESİNİN DEĞERLENDİRİLMESİ

Year 2022, , 486 - 492, 30.09.2022
https://doi.org/10.17343/sdutfd.1163402

Abstract

Amaç
Gıda katkı maddelerinin kullanımı endüstriyel gelişmeyle
birlikte sürekli artmaktadır. Bu çalışmada sık
kullanılan sitrik asit, askorbik asit ve sodyum sitratın
insan lenfosit hücrelerinde DNA üzerine olan etkilerinin
comet metodu ile belirlenmesi amaçlanmıştır.
Gereç ve Yöntem
Herhangi bir bilinen hastalığı ya da sürekli ilaç kullanımı
olmayan, son 6 ayda radyolojik muayene geçirmemiş
ve sigara içmeyen 18 – 45 yaş aralığında 4 erkek
ve 4 kadın gönüllüden kan alınmış ve lenfositler izole
edilerek 50, 150, 300 ve 600 μg/mL dozlarında sitrik
asit, askorbik asit ve sodyum sitrat ile 1 s inkübasyona
bırakılmıştır. Ardından comet metodu uygulanmış ve
kuyruk DNA yüzdesi parametresi DNA hasarının göstergesi
olarak Tek-yönlü Anova ile değerlendirilmiştir.
Bulgular
Sodyum sitrat 300 ve 600 μg/mL dozlarında DNA
hasarında kontrol grubuna kıyasla sınırlı artış görülmektedir.
Bu artış istatistiksel olarak anlamlı değildir
(p>0,05). Sitrik asit 600 μg/mL dozu ise kontrol grubuna
kıyasla oldukça yüksek DNA hasarına sebep
olmuştur (p<0,001). Aynı oranda doz uygulanan askorbik
asit ve sodyum sitrat ile karşılaştırıldığında da
yüksek DNA hasarına sebep olduğu görülmektedir
(p<0,001). Diğer gruplarda DNA hasarında anlamlı bir
değişiklik tespit edilememiştir (p>0,05).
Sonuç
Çalışma sonucunda, denenen dozlarda askorbik asitin
herhangi bir DNA hasarına sebep olmadığı, sitrik
asitin yüksek dozlarda DNA hasarına sebep olduğu
ve sodyum sitratın ise yüksek dozlarda DNA hasarına
sebep olabileceği tespit edilmiştir. Bazı katkı maddeleri
her ne kadar antioksidan olarak sınıflandırılsa da,
yüksek dozlarda DNA hasarına sebep olabilir. Dolayısı
ile bu katkı maddelerini içeren gıdaların tüketiminde
bilinçli ve kontrollü olunması gerekmektedir. Ancak
kesin bir yargıya varmadan önce gıda katkı maddeleri
ile ilgili daha detaylı çalışmalara ihtiyaç duyulmaktadır.

Supporting Institution

Yok

Project Number

Yok

Thanks

Yok

References

  • 1. Carocho M, Barreiro MF, Morales P, Ferreira IC. Adding molecules to food, pros and cons: A review on synthetic and natural food additives. Comprehensive reviews in food science and food safety. 2014;13(4):377-99.
  • 2. Peycheva E, Alexandrova R, Miloshev G. Application of the yeast comet assay in testing of food additives for genotoxicity. LWT-Food Science and Technology. 2014;59(1):510-7.
  • 3. Abd-Elhakim YM, Hashem MM, Anwar A, El-Metwally AE, Abo-El-Sooud K, Moustafa GG, et al. Effects of the food additives sodium acid pyrophosphate, sodium acetate, and citric acid on hemato-immunological pathological biomarkers in rats: Relation to PPAR-α, PPAR-γ and tnfα signaling pathway. Environmental Toxicology and Pharmacology. 2018;62:98-106.
  • 4. Yılmaz S, Ünal F, Yüzbaşıoğlu D, Aksoy H. Clastogenic effects of food additive citric acid in human peripheral lymphocytes. Cytotechnology. 2008;56(2):137-44.
  • 5. Türkoğlu Ş. Genotoxicity of five food preservatives tested on root tips of Allium cepa L. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2007;626(1):4-14.
  • 6. Chen X, Lv Q, Liu Y, Deng W. Effects of the food additive, citric acid, on kidney cells of mice. Biotechnic & Histochemistry. 2015;90(1):38-44.
  • 7. Varvara M, Bozzo G, Celano G, Disanto C, Pagliarone CN, Celano GV. The use of ascorbic acid as a food additive: technical-legal issues. Italian journal of food safety. 2016;5(1).
  • 8. Sammel L, Claus J, Greaser M, Richards M. Investigation of mechanisms by which sodium citrate reduces the pink color defect in cooked ground turkey. Meat science. 2006;72(4):585-95.
  • 9. Lenzi LJ, Lucchesi PM, Medico L, Burgán J, Krüger A. Effect of the food additives sodium citrate and disodium phosphate on Shiga toxin-producing Escherichia coli and production of stx-phages and Shiga toxin. Frontiers in Microbiology. 2016;7:992.
  • 10. Pandir D. DNA damage in human germ cell exposed to the some food additives in vitro. Cytotechnology. 2016;68(4):725-33.
  • 11. Demir E, Kocaoğlu S, Kaya B. Assessment of genotoxic effects of benzyl derivatives by the comet assay. Food and chemical toxicology. 2010;48(5):1239-42.
  • 12. Carocho M, Ferreira IC. A review on antioxidants, prooxidants and related controversy: Natural and synthetic compounds, screening and analysis methodologies and future perspectives. Food and chemical toxicology. 2013;51:15-25.
  • 13. Dosay-Akbulut M. Determination of DNA Damage Caused by Food Additives Using Comet Assay Method: Food Additives DNA damage via comet assay. Progress in Nutrition. 2021;22(4):e2020071.
  • 14. Raya SA, Aboul-Enein AM, El-Nikeety MM, Mohamed RS, Abdelwahid WM. In Vivo comet assay of food additives’ combinations and their effects on biochemical parameters in albino rats. Biointerface Research in Applied Chemistry. 2020;24:9170-83.
  • 15. Ali MY, Hassan GM, Hassan AMS, Mohamed ZA, Ramadan MF. In vivo genotoxicity assessment of sunset yellow and sodium benzoate in female rats. Drug and Chemical Toxicology. 2020;43(5):504-13.
  • 16. Shimada C, Kano K, Sasaki YF, Sato I, Tsudua S. Differential colon DNA damage induced by azo food additives between rats and mice. The Journal of toxicological sciences. 2010;35(4):547-54.
  • 17. Dorier M, Béal D, Marie-Desvergne C, Dubosson M, Barreau F, Houdeau E, et al. Continuous in vitro exposure of intestinal epithelial cells to E171 food additive causes oxidative stress, inducing oxidation of DNA bases but no endoplasmic reticulum stress. Nanotoxicology. 2017;11(6):751-61.
  • 18. Abo-EL-Sooud K, Hashem MM, Badr YA, Eleiwa MM, Gab-Allaha AQ, Abd-Elhakim YM, et al. Assessment of hepato-renal damage and genotoxicity induced by long-term exposure to five permitted food additives in rats. Environmental Science and Pollution Research. 2018;25(26):26341-50.
  • 19. Mpountoukas P, Vantarakis A, Sivridis E, Lialiaris T. Cytogenetic study in cultured human lymphocytes treated with three commonly used preservatives. Food and Chemical Toxicology. 2008;46(7):2390-3.
  • 20. Abd‐Elhakim YM, Anwar A, Hashem MM, Moustafa GG, Abo‐El‐Sooud K. Sodium acetate, sodium acid pyrophosphate, and citric acid impacts on isolated peripheral lymphocyte viability, proliferation, and DNA damage. Journal of Biochemical and Molecular Toxicology. 2018;32(8):e22171.
  • 21. Mamur S, Yüzbaşıoğlu D, Unal F, Aksoy H. Genotoxicity of food preservative sodium sorbate in human lymphocytes in vitro. Cytotechnology. 2012;64(5):553-62.
  • 22. Ataseven N, Yüzbaşıoğlu D, Keskin AÇ, Ünal F. Genotoxicity of monosodium glutamate. Food and Chemical Toxicology. 2016;91:8-18.
  • 23. OECD. Test No. 489: In Vivo Mammalian Alkaline Comet Assay. Paris: OECD Publishing; 2016.
  • 24. Gyori BM, Venkatachalam G, Thiagarajan P, Hsu D, Clement M-V. OpenComet: an automated tool for comet assay image analysis. Redox biology. 2014;2:457-65.
  • 25. IBMCorp. IBM SPSS Statistics for Windows, Version 20.0. Armonk, NY: IBM Corp.; 2011.
  • 26. Lerda D, Biaggi Bistoni M, Peralta N, Ychari S, Vazquez M, Bosio G. Fumonisins in foods from Cordoba (Argentina), presence and genotoxicity. Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association. 2005;43(5):691-8.
  • 27. Yilmaz S, Ünal F, Yüzbaşıoğlu D, Celik M. DNA damage in human lymphocytes exposed to four food additives in vitro. Toxicology and Industrial Health. 2014;30(10):926-37.
  • 28. Mamur S, Yüzbaşıoğlu D, Ünal F, Yılmaz S. Does potassium sorbate induce genotoxic or mutagenic effects in lymphocytes? Toxicology in vitro. 2010;24(3):790-4.
  • 29. Zengin N, Yüzbaşıoğlu D, Ünal F, Yılmaz S, Aksoy H. The evaluation of the genotoxicity of two food preservatives: Sodium benzoate and potassium benzoate. Food and Chemical Toxicology. 2011;49(4):763-9.
  • 30. Kaya FF, Topaktaş M. Genotoxic effects of potassium bromate on human peripheral lymphocytes in vitro. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2007;626(1-2):48-52.

GENOTOXICITY EVALUATION OF CITRIC ACID, ASCORBIC ACID AND SODIUM CITRATE USED AS FOOD ADDITIVES IN HUMAN LYMPHOCYTES

Year 2022, , 486 - 492, 30.09.2022
https://doi.org/10.17343/sdutfd.1163402

Abstract

Objective
The use of food additives is constantly increasing
with industrial development. In this study, we aimed
to determine the effects of commonly used citric acid,
ascorbic acid, and sodium citrate on DNA in human
lymphocytes by the comet assay.
Material and Method
Blood samples were collected from 4 male and 4 female
volunteers, aged 18 to 45 years, who had no known
disease or continuous drug use, had not undergone
radiological examination in the past 6 months, and
were nonsmokers. Lymphocytes were isolated and
incubated for 1 h with citric acid, ascorbic acid, and
sodium citrate at doses of 50, 150, 300, and 600 μg/
mL. Subsequently, the Comet assay was applied and
the tail DNA percentage was evaluated as evidence of
DNA damage using One-way Anova.
Results
A limited increase in DNA damage was observed at
sodium citrate doses of 300 and 600 μg/ml compared
to the control group. This increase was not statistically
significant (p>0.05). The dose of 600 μg/mL citric acid
caused significantly higher DNA damage compared to
the control group (p<0.001). Likewise, when compared
to the same dose of ascorbic acid and sodium citrate,
dose of 600 μg/mL citric acid caused significantly
higher DNA damage (p<0.001). No significant change
in DNA damage was observed in any other groups.
Conclusion
As a result, it was found that ascorbic acid did not
cause DNA damage at all doses, citric acid caused
DNA damage at high doses, and sodium citrate might
cause DNA damage at high doses. Although some
additives are classified as antioxidants, they may
cause DNA damage at high doses. Therefore, it is
necessary to be aware and controlled when consuming
foods containing these additives. However, more
detailed studies on food additives are needed before a
definitive conclusion can be drawn.

Project Number

Yok

References

  • 1. Carocho M, Barreiro MF, Morales P, Ferreira IC. Adding molecules to food, pros and cons: A review on synthetic and natural food additives. Comprehensive reviews in food science and food safety. 2014;13(4):377-99.
  • 2. Peycheva E, Alexandrova R, Miloshev G. Application of the yeast comet assay in testing of food additives for genotoxicity. LWT-Food Science and Technology. 2014;59(1):510-7.
  • 3. Abd-Elhakim YM, Hashem MM, Anwar A, El-Metwally AE, Abo-El-Sooud K, Moustafa GG, et al. Effects of the food additives sodium acid pyrophosphate, sodium acetate, and citric acid on hemato-immunological pathological biomarkers in rats: Relation to PPAR-α, PPAR-γ and tnfα signaling pathway. Environmental Toxicology and Pharmacology. 2018;62:98-106.
  • 4. Yılmaz S, Ünal F, Yüzbaşıoğlu D, Aksoy H. Clastogenic effects of food additive citric acid in human peripheral lymphocytes. Cytotechnology. 2008;56(2):137-44.
  • 5. Türkoğlu Ş. Genotoxicity of five food preservatives tested on root tips of Allium cepa L. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2007;626(1):4-14.
  • 6. Chen X, Lv Q, Liu Y, Deng W. Effects of the food additive, citric acid, on kidney cells of mice. Biotechnic & Histochemistry. 2015;90(1):38-44.
  • 7. Varvara M, Bozzo G, Celano G, Disanto C, Pagliarone CN, Celano GV. The use of ascorbic acid as a food additive: technical-legal issues. Italian journal of food safety. 2016;5(1).
  • 8. Sammel L, Claus J, Greaser M, Richards M. Investigation of mechanisms by which sodium citrate reduces the pink color defect in cooked ground turkey. Meat science. 2006;72(4):585-95.
  • 9. Lenzi LJ, Lucchesi PM, Medico L, Burgán J, Krüger A. Effect of the food additives sodium citrate and disodium phosphate on Shiga toxin-producing Escherichia coli and production of stx-phages and Shiga toxin. Frontiers in Microbiology. 2016;7:992.
  • 10. Pandir D. DNA damage in human germ cell exposed to the some food additives in vitro. Cytotechnology. 2016;68(4):725-33.
  • 11. Demir E, Kocaoğlu S, Kaya B. Assessment of genotoxic effects of benzyl derivatives by the comet assay. Food and chemical toxicology. 2010;48(5):1239-42.
  • 12. Carocho M, Ferreira IC. A review on antioxidants, prooxidants and related controversy: Natural and synthetic compounds, screening and analysis methodologies and future perspectives. Food and chemical toxicology. 2013;51:15-25.
  • 13. Dosay-Akbulut M. Determination of DNA Damage Caused by Food Additives Using Comet Assay Method: Food Additives DNA damage via comet assay. Progress in Nutrition. 2021;22(4):e2020071.
  • 14. Raya SA, Aboul-Enein AM, El-Nikeety MM, Mohamed RS, Abdelwahid WM. In Vivo comet assay of food additives’ combinations and their effects on biochemical parameters in albino rats. Biointerface Research in Applied Chemistry. 2020;24:9170-83.
  • 15. Ali MY, Hassan GM, Hassan AMS, Mohamed ZA, Ramadan MF. In vivo genotoxicity assessment of sunset yellow and sodium benzoate in female rats. Drug and Chemical Toxicology. 2020;43(5):504-13.
  • 16. Shimada C, Kano K, Sasaki YF, Sato I, Tsudua S. Differential colon DNA damage induced by azo food additives between rats and mice. The Journal of toxicological sciences. 2010;35(4):547-54.
  • 17. Dorier M, Béal D, Marie-Desvergne C, Dubosson M, Barreau F, Houdeau E, et al. Continuous in vitro exposure of intestinal epithelial cells to E171 food additive causes oxidative stress, inducing oxidation of DNA bases but no endoplasmic reticulum stress. Nanotoxicology. 2017;11(6):751-61.
  • 18. Abo-EL-Sooud K, Hashem MM, Badr YA, Eleiwa MM, Gab-Allaha AQ, Abd-Elhakim YM, et al. Assessment of hepato-renal damage and genotoxicity induced by long-term exposure to five permitted food additives in rats. Environmental Science and Pollution Research. 2018;25(26):26341-50.
  • 19. Mpountoukas P, Vantarakis A, Sivridis E, Lialiaris T. Cytogenetic study in cultured human lymphocytes treated with three commonly used preservatives. Food and Chemical Toxicology. 2008;46(7):2390-3.
  • 20. Abd‐Elhakim YM, Anwar A, Hashem MM, Moustafa GG, Abo‐El‐Sooud K. Sodium acetate, sodium acid pyrophosphate, and citric acid impacts on isolated peripheral lymphocyte viability, proliferation, and DNA damage. Journal of Biochemical and Molecular Toxicology. 2018;32(8):e22171.
  • 21. Mamur S, Yüzbaşıoğlu D, Unal F, Aksoy H. Genotoxicity of food preservative sodium sorbate in human lymphocytes in vitro. Cytotechnology. 2012;64(5):553-62.
  • 22. Ataseven N, Yüzbaşıoğlu D, Keskin AÇ, Ünal F. Genotoxicity of monosodium glutamate. Food and Chemical Toxicology. 2016;91:8-18.
  • 23. OECD. Test No. 489: In Vivo Mammalian Alkaline Comet Assay. Paris: OECD Publishing; 2016.
  • 24. Gyori BM, Venkatachalam G, Thiagarajan P, Hsu D, Clement M-V. OpenComet: an automated tool for comet assay image analysis. Redox biology. 2014;2:457-65.
  • 25. IBMCorp. IBM SPSS Statistics for Windows, Version 20.0. Armonk, NY: IBM Corp.; 2011.
  • 26. Lerda D, Biaggi Bistoni M, Peralta N, Ychari S, Vazquez M, Bosio G. Fumonisins in foods from Cordoba (Argentina), presence and genotoxicity. Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association. 2005;43(5):691-8.
  • 27. Yilmaz S, Ünal F, Yüzbaşıoğlu D, Celik M. DNA damage in human lymphocytes exposed to four food additives in vitro. Toxicology and Industrial Health. 2014;30(10):926-37.
  • 28. Mamur S, Yüzbaşıoğlu D, Ünal F, Yılmaz S. Does potassium sorbate induce genotoxic or mutagenic effects in lymphocytes? Toxicology in vitro. 2010;24(3):790-4.
  • 29. Zengin N, Yüzbaşıoğlu D, Ünal F, Yılmaz S, Aksoy H. The evaluation of the genotoxicity of two food preservatives: Sodium benzoate and potassium benzoate. Food and Chemical Toxicology. 2011;49(4):763-9.
  • 30. Kaya FF, Topaktaş M. Genotoxic effects of potassium bromate on human peripheral lymphocytes in vitro. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2007;626(1-2):48-52.
There are 30 citations in total.

Details

Primary Language Turkish
Subjects Clinical Sciences
Journal Section Research Articles
Authors

Dilek Aşcı Çelik 0000-0002-2914-4695

Vehbi Atahan Toğay 0000-0003-4722-3845

Gülçin Yavuz Türel 0000-0001-9481-4475

Nurten Özçelik 0000-0003-2326-6090

Project Number Yok
Publication Date September 30, 2022
Submission Date August 17, 2022
Acceptance Date September 2, 2022
Published in Issue Year 2022

Cite

Vancouver Aşcı Çelik D, Toğay VA, Yavuz Türel G, Özçelik N. GIDA KATKI MADDESİ OLARAK KULLANILAN SİTRİK ASİT, ASKORBİK ASİT VE SODYUM SİTRATIN İNSAN LENFOSİT HÜCRELERİNDE GENOTOKSİSİTESİNİN DEĞERLENDİRİLMESİ. Med J SDU. 2022;29(3):486-92.

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