Gıdalarda koruyucu katkı maddesi olarak kullanılan sodyum benzoat ve potasyum sorbat karışımının genotoksik potansiyelinin mikronukleus testi ile belirlenmesi

Öz

Sodyum benzoat (SB) ve potasyum sorbat (PS), gıdalarda koruyucu katkı maddesi olarak yaygın bir şekilde kullanılmaktadır.  Bu çalışmada SB ve PS karışımının genotoksik potansiyeli insan lenfositlerinde mikronukleus (MN) testi ile değerlendirilmiştir.  SB+PS karışımının dokuz farklı kombinasyonu (12.5 SB+125 PS ile 50 SB+500 PS μg/mL aralığında) kültüre alınmış insan lenfositlerine 48 saat uygulanmıştır.  MN testi sonuçlarına göre; SB+PS karışımının uygulanan tüm konsantrasyonlarda mikronukleus frekansını kontrole kıyasla anlamlı düzeyde artırdığı belirlenmiştir.  Buna karşın, SB+PS karışımının çalışılan konsantrasyonlarda hücre siklusu belirteci olan nükleeer bölünme indeksini (NBİ) anlamlı düzeyde etkilemediği tespit edilmiştir.  Elde edilen sonuçlar, SB+PS karışımının in vitro koşullarda MN testi ile insan lenfositlerinde klastojenik ve/veya anöjenik etkileri olabileceğini göstermektedir.

Anahtar Kelimeler

• Gıda koruyucusu,sodyum benzoat,potasyum sorbat,insan lenfositleri,mikronukleus,nükleer bölünme indeksi

Determination of genotoxic potential of food preservative mixture of sodium benzoate and potassium sorbate by micronucleus test

Öz

Sodium benzoate (SB) and potassium sorbate (PS) are widely used as preservative additive in food.  The present study evaluates genotoxic potential of a mixture of SB and PS in cultured human lymphocytes by using micronucleus (MN) test.  To assess genotoxic effect, human lymphocytes were treated with nine different combinations of SB+PS (between 12.5 SB+125 PS and 50 SB+500 PS μg/mL) for 48 h.  MN test revealed that the mixture of SB+PS treatment significantly increased the MN frequency at all concentrations compared to vehicle control.  On the contrary, the mixture of SB+PS did not significantly affect the nuclear division index (NDI) that is cell cycle biomarker.  The results obtained show that the mixture of SB+PS probably induce clastogenic and/or aneugenic effects in human peripheral blood lymphocytes in vitro by MN test. 

Anahtar Kelimeler

• Food preservative,sodium benzoate,potassium sorbate,human lymphocytes,micronucleus,nuclear division index

Kaynakça

1. Saldamlı, İ. and Uygun, Ü. Gıda Katkı Maddeleri., Saldamlı, İ. (Editör). Gıda Kimyası, Dördüncü Baskı. Ankara. Hacettepe Üniversitesi Yayınları, 625-668, (2014).
2. Altuğ, T., Gıda Katkı Maddeleri., T. Altuğ (Editör). Gıda katkı maddeleri, Üçüncü Baskı. İzmir. Kan Yılmaz Matbaacılık, 1-16, (2009).
3. Shariati, M., Matin, S., Ghanei, M., Kalani, N., Nitrate, sorbate and benzoate a jusification for the conflicting results of epidemiological studies of cancer in nutrition, Journal of Fundamental and Applied Sciences, 8, 3S, 2273-2285, (2016).
4. Türk Gıda Kodeksi, Gıda Katkı Maddeleri Yönetmeliği, Gıda, Tarım ve Hayvancılık Bakanlığı, T.C. Resmi Gazete, Sayı: 28157, (2011).
5. Türk Gıda Kodeksi, Gıda Katkı Maddeleri Yönetmeliği, Gıda, Tarım ve Hayvancılık Bakanlığı, T.C. Resmi Gazete, Sayı: 28693, (2013).
6. Ova, G., Koruyucular., T. Altuğ (Editör), Gıda katkı maddeleri, Üçüncü Baskı. İzmir. Kan Yılmaz Matbaacılık, 105-156, (2009).
7. FDA U. GRAS (Generally Recognized As Safe) food ingredients: Benzoic acid and sodium benzoate, Food and Drug Administration, Washington, DC, (1972).
8. Lück E., Food applications of sorbic acid and its salts, Food Additives and Contaminants, 7, 5, 711-5, (1990).

9. Fenech, M., Knasmueller, S., Bolognesi, C., Bonassi, S., Holland, N., Migliore, L., Palitti, F., Natarajan, A.T. and Kirsch-Volders, M., Molecular mechanisms by which in vivo exposure to exogenous chemical genotoxic agents can lead to micronucleus formation in lymphocytes in vivo and ex vivo in humans, Mutation Research, 770, 12-25, (2016).
10. Kirsch-Volders, M., Plas, G., Elhajouji, A., Lukamowicz, M., Gonzalez, L., Loock, K.V. and Decordier, I., The in vitro MN assay in 2011: origin and fate, biological significance, protocols, high throughput methodologies and toxicological relevance, Archives Toxicology, 85, 8, 873-899, (2011).
11. Parry, J.M. and Kirsch-Volders, M., Special issue on in vitro MN trial, Mutation Research, 702, 132-134, (2010).
12. Chandirasekar, R., Kumar, B.L., Sasikala, K., Jayakumar, R., Suresh, K., Venkatesan, R., Jacob, R., Krishnapriya, E.K., Kavitha, R. and Ganesh, G.K., Assessment of genotoxic and molecular mechanisms of cancer risk in smoking and smokeless tobacco users, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 767, 21-27, (2014).
13. Fenech, M., The in vitro micronucleus technique. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 455, 1, 81-95, (2000).
14. Huerta, I., Barasoain, M., Télez, M., Longa, M., Muga, J., Barrenetxea, G., Ortiz-Lastrad, E., Gonzálezd, J., Criado, B. and Arrieta, I., Genotoxic evaluation of five Angiotesin II receptor blockers: In vivo and in vitro micronucleus assay, Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 767, 1-7, (2014).
15. Bhatia, A. and Kumar, Y., Cancer cell micronucleus: an update on clinical and diagnostic applications, Acta Pathologica, Microbiologica, et Immunologica Scandinavica, 121, 7, 569-81, (2013).
16. Bonassi, S., El-Zein, R., Bolognesi, C. and Fenech, M., Micronuclei frequency in peripheral blood lymphocytes and cancer risk: evidence from human studies, Mutagenesis, 26, 1, 93-100, (2011).
17. Sodium Benzoate, https://www.google.com.tr/search?q=sodyum+benzoat&safe= active&rlz=1C1EJFA_enTR729TR729&source=lnms&tbm=isch&sa=X&ved=0ahUKEwiOk4eUxNnYAhUkQZoKHcxhALgQ_AUICigB&biw=1600&bih=794#imgrc=HGBIDqAPSs-_rM, (06.03.2018)
18. Potassium sorbate analytical standard, https://www.sigmaaldrich.com/catalog/ product/supelco/47848?lang=en®ion=TR, (06.03.2018)
19. Zengin, N., Yüzbaşıoğlu, D., Ünal, F., Yılmaz, S. and Aksoy, H., The evaluation of the genotoxicity of two food preservatives: sodium benzoate and potassium benzoate, Food and Chemical Toxicology, 49, 4, 763-769, (2011).
20. Mamur, S., Yüzbaşıoğlu, D., Ünal, F. and Yılmaz, S., Does potassium sorbate induce genotoxic or mutagenic effects in lymphocytes?, Toxicology In Vitro, 24, 790–794, (2010).
21. Palus, J., Rydzynski, K., Dziubaltowska, E., Wyszynska, K., Natarajan, A.T. and Nilsson, R., Genotoxic effects of occupational exposure to lead and cadmium, Mutation Research, 540, 19-28, (2003).
22. Surrales, J., Xamena, N., Creus, A., Catalan, J., Norppa, H. and Marcos, R., Induction of micronuclei by five pyrethroid insecticides in whole blood and isolated human lymphocytes cultures, Mutation Research, 341, 169-184, (1995).
23. Shahmohammadi, M., Javadi, M. and Nassiri-Asl, M., An overview on the Effects of Sodium Benzoate as a Preservative in Food Products, Biotechnology and Health Sciences, 3, 3, e35084, (2016). DOI: 10.17795/bhs-35084.
24. Erickson M.C. and Doyle, M.P., The challenges of eliminating or substituting antimicrobial preservatives in foods, Annual Review of Food Science and Technology, 8, 371-90, (2017).
25. Yüzbaşıoğlu, D., Zengin, N. and Ünal, F., Gıda koruyucuları ve genotoksisite testleri, Gıda/The Journal of Food, 39, 3, 179-186, (2014).
26. Yılmaz, S., Ünal, F., Aksoy, H., Yüzbaşıoğlu, D. and Çelik, M., Cytogenetic effects of citric acid and benzoic acid on Allium chromosomes, Fresenius Environmental Bulletin, 17, 8A, 1029-1037, (2008).
27. Yılmaz, S., Ünal, F. and Yüzbaşıoğlu, D., The in vitro genotoxicity of benzoic acid in human peripheral blood lymphocytes, Cytotechnology, 60, 1-3, 55-61, (2009).
28. Yılmaz, S., Ünal, F., Yüzbaşıoğlu, D. and Çelik, M., DNA damage in human lymphocytes exposed to four food additives in vitro, Toxicology and Industrial Health, 30, 10, 926-37, (2014).
29. Çulcu, T., Sözen, E. and Tüylü, B. A. Determination of genotoxicants-induced DNA damage by using RAPD-PCR in human peripheral blood lymphocytes, Fresenius Environmental Bulletin, 19, 10, 2205-2209, (2010).
30. Mamur, S., Yüzbaşıoğlu, D., Ünal, F. and Aksoy, H., Genotoxicity of food preservative sodium sorbate in human lymphocytes in vitro, Cytotechnology, 64, 553-562, (2012).
31. Özdemir, H., Turhan, A.B. and Arıkoğlu, H., Potasyum sorbat, sodyum benzoat ve sodyum nitrit’in genotoksik etkilerinin araştırılması, European Journal of Basic Medical Sciences, 2, 2, 34-40, (2012).
32. Türkoğlu, Ş., Evaluation of genotoxic effects of sodium propionate, calcium propionate and potassium propionate on the root meristem cells of Allium cepa, Food and Chemical Toxicology, 46, 6, 2035-2041, (2008).
33. Ataseven, N., Yüzbaşıoğlu, D., Keskin, A.Ç. and Ünal, F., Genotoxicity of monosodium glutamate, Food and Chemical Toxicology, 91, 8-18, (2016).
34. García-Quispes, W.A., Pastor, S., Biarnes, P.G.F., Castell, J., Velazquez, A. and Marcos, R., Influence of DNA-repair gene variants on the micronucleus frequency in thyroid cancer patients, Mutation Research, 750, 34-39, (2013).
35. Terradas, M., Martin, M., Tusell, L. and Genesca, A., Genetic activities in micronuclei: is the DNA entrapped in micronuclei lost for the cell?, Mutation Research, 705, 60-67, (2010).
36. Eastmond, D.A. and Tucker, J.D., Kinetochore localization in micronucleated cytokinesis-blocked Chinese hamster ovary cells: a new and rapid assay for identifying aneuploidy-inducing agents, Mutation Research, 224, 517-525, (1989).
37. Fenech, M., Cytokinesis-block micronucleus cytome assay, Nature Protocols, 2, 5, 1084-1104, (2007).
38. Ionescu, M.E., Ciocirlan, M., Becheanu, G., Nicolaie, T., Ditescu, C., Teiusanu, A.G., Gologan, S.I. and Arbanas, T., Nuclear division ındex may predict neoplastic colorectal lesions, Maedica (Buchar), 6, 3, 173-178, (2011).
39. Mpountoukas, P., Vantarakis, A., Sivridis, E. and Lialiaris, T., Cytogenetic study in cultured human lymphocytes treated with three commonly used preservatives, Food and Chemical Toxicology, 46, 7, 2390-2393, (2008).
40. Pandir, D., DNA damage in human germ cell exposed to the some food additives in vitro, Cytotechnology, 68, 725-733, (2016).
41. Stanojevic, D., Comic, I., Stefanovıc O. and Solujıc-Sukdolak S.L., Antimicrobial effects of sodium benzoate, sodium nitrite and potassium sorbate and their synergistic action in vitro, Bulgarian Journal of Agricultural Science, 15, 307-311, (2009).
42. Pérez-Prior, M.T., Manso, J.A., García-Santos, M. del P., Calle, E. and Casado, J., Alkylating potential of potassium sorbate, Journal of Agricultural and Food Chemistry, 53, 6, 10244-10247, (2005).
43. Weber, G.F., DNA damaging drugs, Molecular Therapies of Cancer, 9-112, (2015). DOI 10.1007/978-3-319-13278-5_2.
44. Fenech, M. and Bonassi, S., The effect of age, gender, diet and lifestyle on DNA damage measured using micronucleus frequency in human peripheral blood lymphocytes, Mutagenesis, 26, 43–49, (2011).
45. Mørck, T.A., Loock, K.V., Poulsen, M.B., Siersma, V.D., Nielsen, J.K.S., Hertel, O., Kirsch-Volders, M. and Knudsen, L.E., Micronucleus frequency in Danish school children and their mothers from the DEMOCOPHES population, Mutagenesis, 31, 1-8, (2016).