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

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.

Keywords

• Comet Assay
• DNA Damage
• E300
• E330
• E331

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

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.

Keywords

• Comet Metodu
• DNA Hasarı
• E300
• E330
• E331

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