Sains Malaysiana 50(5)(2021): 1343-1356


Effect of Food Colorants Supplementation on Reactive Oxygen Species, Antioxidant Vitamins Level and DNA Damage

(Kesan Tambahan Pewarna Makanan pada Spesies Oksigen Reaktif, Tahap Antioksidan Vitamin dan Kerosakan DNA)




1Department of Nutrition, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia


2Sub Campus Burewala-Vehari, University of Agriculture Faisalabad, Punjab, Pakistan


3Research Centre of Excellence, Nutrition and Non-Communicable Diseases (RCOE NNCD), Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia


4Institute of Pharmacy, Pharmacology, and Physiology, University of Agriculture Faisalabad, Punjab, Pakistan


5Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia


Diserahkan: 29 Mac 2020/Diterima: 8 Oktober 2020  



There are various undesirable products generated from endogenous aerobic metabolism such as reactive oxygen species (ROS). Physiological and biochemical lesions are caused by ROS and which give rise to oxidative damages towards DNA, proteins and lipids which ultimately lead to cell death. This study was aimed to examine the effect of oral administration of food colorants (tartrazine and curcumin) on the oxidants and antioxidants level in blood and fecal of rats after 15, 30, and 45 days. Two doses were used based on the admissible daily intake (ADI) of 9.6 and 96 (high) mg/kg/body weight for tartrazine, 3.85 and 38.5 6 mg/kg/body weight for curcumin. The results showed that oral administration of tartrazine had significantly increased the total oxidant level, arylesterase, glutathione reductase, and MDA whereas there was significantly decreased the total antioxidants level, catalase, glutathione peroxidase in plasma and fecal after 30 and 45 days. Vitamin E and C were decreased in plasma. Fecal showed high level of vitamin A. High dose of tartrazine caused alteration in the aldehyde reactive probe (ARP) sites of DNA showing the DNA damage. After 45 day, significant increment was observed in the level of AST in low and high curcumin treated group. Whereas, elevations of arylestrase were seen in high curcumin group after 45 day. High dose of curcumin significantly (P≤ 0.05) decreased the concentration of vitamin C after 45 days of treatment and increased the level of vitamin E in plasma of treated groups after 30 and 45 days of treatment. The present study showed that the ADI and doses up to 10 times higher than ADI showed negative effects on antioxidant level and demonstrated the importance of using appropriate doses of food colorants such as tartrazine and curcumin in different processed food products.


Keywords: Antioxidants; arylesterase; food colorants; glutathione; lipid peroxidation; ROS



Terdapat pelbagai produk yang tidak diingini yang dijanakan daripada metabolisme aerobik endogen seperti spesies oksigen reaktif (ROS). Lesi fisiologikal dan biokimia disebabkan oleh ROS yang mengakibatkan peningkatan kepada kerosakan oksidatif terhadap DNA, protein dan lipid yang akhirnya membawa kepada kematian sel. Kajian ini bertujuan untuk meneliti kesan pelaksanaan oral pewarna makanan (tartrazin dan kurkumin) ke atas tahap oksidan dan antioksidan dalam darah dan najis tikus selepas 15, 30 dan 45 hari. Dua dos telah digunakan berdasarkan pengambilan harian yang dibenarkan (ADI) iaitu 9.6 dan 96 (tinggi) mg/kg/berat badan bagi tartrazin, 3.85 dan 38.5 6 mg/kg/berat badan bagi curcumin. Keputusan menunjukkan bahawa pelaksanaan oral tartrazin telah meningkat secara signifikan total tahap oksidan, arilestras, glutation reduktase dan MDA manakala terdapat penurunan yang signifikan total tahap oksidan, katalase, glutation peroksidase dalam  plasma dan najis selepas 30 dan 45 hari. Vitamin E dan C telah menurun dalam plasma. Najis menunjukkan tahap vitamin A yang tinggi. Dos tartrazin yang tinggi yang menyebabkan pengubahan dalam tapak prob reaktif aldehid (ARP) DNA menunjukkan kerosakan DNA. Selepas 45 hari, kenaikan yang signifikan telah dikesan dalam tahap AST dalam kumpulan terawat rendah dan tinggi kurkumin. Manakala, elevasi arilestras telah kelihatan dalam kumpulan tinggi kurkumin selepas 45 hari. Dos tinggi kurkumin secara signifikan (P≤ 0.05) telah menurunkan kepekatan vitamin C selepas 45 hari rawatan dan meningkatkan tahap vitamin E dalam plasma kumpulan kawalan selepas 30 dan 45 hari rawatan. Kajian ini menunjukkan bahawa ADI dan dos sehingga 10 kali lebih tinggi daripada ADI menunjukkan kesan negatif ke atas tahap antioksidan dan ia memaparkan kepentingan untuk menggunakan dos pewarna makanan yang sesuai seperti tartrazin dan  kurkumin dalam produk makanan diproses yang berbeza.


Kata kunci: Arilesteras; glutation; pewarna makanan; pemperoksidaan lipid; ROS




Abd El-Galeel, M.A.S. 2002. Solubility and stability of natural food colorants in microemulsions.  University Bonn, Ph.D. Thesis (Unpublished).

Abdel-Aziz, H.M., Alazouny, Z.M., Abdelfadeel, K.F. & Abohashem, A.A. 2019. Effect of tartrazine on thyroid gland of male rat and ameliorating role of curcumin (histological and immunohistochemical study). Journal of Biochemistry and Cell Biology 2(111): 2.

Alam, M.N., Nusrat, J.B. & Rafiquzzaman, M.D. 2012. Review on in vivo and in vitro methods evaluation of antioxidant activity. Saudi Pharmaceutical Journal 21: 143-152.

Amani, A.R., Somchit, M.N., Konting, M.M. & Kok, L. 2010. Vitamin E and curcumin intervention on lipid-peroxidation and antioxidant defense system. Journal of American Science 6: 52-62.

Amin, K., Hameid, H.A. & Elsttar, A.A. 2010. Effect of food azo dyes tartrazine and carmoisine on biochemical parameters related to renal, hepatic function and oxidative stress biomarkers in young male rats. Food and Chemical Toxicology 48(10): 2994-2999.

Antunes, L.M., Araujo, M.C., Dias, F.L. & Takahashi, C.S. 1999. Modulatory effects of curcumin on the chromosomal damage induced by doxorubicin in Chinese hamster ovary cells. Teratogen Carcinogen Mutagen 19: 1-8.

Aquilano, K., Baldelli, S. & Ciriollo, M.R. 2014. Glutathione: New roles in redox signaling for an old antioxidant. Front Pharmacology 5: 196.

Ashida, H., Hashimoto, T., Tsuji, S., Kanazawa, K. & Danno, G.I. 2000. Synergistic effects of food colors on the toxicity of 3-amino-1, 4-dimethyl-5H-pyrido [4, 3-b] indole (Trp-P-1) in primary cultured rat hepatocytes. Journal of Nutritional Science and Vitaminology 46(3): 130-136.

Babbs, C.F. 1990. Free radicals and the etiology of colon cancer. Free Radical Biology and Medicine 8: 191-200.

Babior, B.M. 1978. Oxygen-dependent microbial killing by phagocytes. New England Journal of Medicine 298: 659-668.

Bansal, A.K., Bansal, M., Soni, G. & Bhatnagar, D. 2005. Modulation of N-nitrosodiethylamine (NDEA) induced oxidative stress by vitamin E in rat erythrocytes. Human Experimental Toxicology 24: 297-302.

Bergmeier, C., Siekmeier, R. & Gross, W. 2004. Distribution spectrum of paraoxonase activity in HDL fractions. Clinical Chemistry 50(12): 2309-2315.

Bergmeyer, H.U. 1965. Methods of Enzyme Analysis. Cambridge: Academic Press. p. 682.

Bianchi, M.A., Francesca, S., Daniele, D.R., Silvia, V., Nicoletta, P., Laura, F., Maria, L.C., Claudia, P., Annamaria, B., Ivana, Z. & Furio, B. 2010. Ability of a high-total antioxidant capacity diet to increase stool weight and bowel antioxidant status in human subjects. British Journal of Nutrition 104(10): 1500-1507.

Blasiak, J., Trzeciak, A., Malecka-Panas, E., Drzewoski, J., Iwanienko, T., Szumiel, I. & Wojewodzka, M. 1999. DNA damage and repair in human lymphocytes and gastric mucosa cells exposed to chromium and curcumin. Teratogen Carcinogen Mutagen 19: 19-31.

Borzelleca, J.F. & Hallagan, J.B. 1988. A chronic toxicity/carcinogenicity study of FD & C Yellow No. 5 (tartrazine) in mice. Food Chem. Toxicol. 26(3): 189-194.

Cemek, M., Büyükokuroğlu, M.E., Sertkaya, F., Alpdağtaş, S., Hazini, A., Önül, A. & Göneş, S. 2014. Effects of food color additives on antioxidant functions and bioelement contents of liver, kidney and brain tissues in rats. Journal of Food and Nutrition Research 2(10): 686-691.

Chung, K.T. & Fulk, G.E. 1981. Mutagenicity testing of some commonly used dyes. Applied and Environmental Microbiology 42(4): 641-648.

Clauss, M., Wittenbrink, M.M., Castell, J.C., Kienzle, E., Dierenfeld, E.S., Flach, E.J. & Hatt, J.M. 2008. Quantification of enterobacteriaceae in faeces of captive black rhinoceros (Diceros bicornis) in relation to dietary tannin supplementation. Journal of Animal Physiology and Animal Nutrition 92(1): 29-34.

Craven, P.A., Fanstiel, J.P. & Derubertis, F.R. 1986. Role of reactive oxygen metabolites in bile salt stimulation of colonic epithelial proliferation. Journal of Clinical Investigations 77: 850-859.

Cruz, T., Galvez, J., Ocete, M.A., Crespo, M.E., Sanchez de Medina, L.H.F. & Zarzuelo, A. 1998. Oral administration of rutoside can ameliorate inflammatory bowel disease in rats. Life Science 62: 687-695.

Demirkol, O., Zhang, X. & Ercal, N. 2012. Oxidative effects of tartrazine (CAS No. 1934-21-0) and new coccin (CAS No. 2611-82-7) azo dyes on CHO cells. Journal Fur Verbraucherschutz Und Lebensmittelsicherhei 7(3): 229-236.

Durrington, P.N., Bhatnagar, D., Mackness, M.I., Morgan, J., Julier, K., Khan, M.A. & France, M. 2001. An omega-3 polyunsaturated fatty acid concentrate administered for one year decreased triglycerides in simvastatin treated patients with coronary heart disease and persisting hypertriglyceridaemia. Heart 85(5): 544-548.

El-Wahab, H.M.F.A. & Moram, G.S.E.D. 2013. Toxic effects of some synthetic food colorants and/or flavor additives on male rats. Toxicology and Industrial Health 29(2): 224-232.

Erel, O. 2005. A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry 38: 1103-1111.

Erel, O. 2004. A novel automated method to measure total antioxidant response against potent free radical reactions. Clinical Biochemistry 37: 112-119.

Erhardt, J.G., Lim, S.S. & Bode, J.C. 1997. A diet rich in fat and poor in dietary fiber increases the in vitro formation of reactive oxygen species in human feces. Journal of Nutrition 127: 706-709.

FAO/WHO. 1999. Evaluation of certain food additives and contaminants (Fifty-first report of the Joint FAO/WHO Expert Committee on Food Additives). WHO Technical Report Series, No. 891.

Farombi, E.O., Abarikwu, S.O., Adedara, I.A. & Oyeyemi, M.O. 2007. Curcumin and kolaviron ameliorate di-n- butylphthalate-induced testicular damage in rats. Basic Clinical Pharmacology and Toxicology 100(1): 43-48.

FDA. 2005. Guidance for Industry: Estimating the Maximum Safe Starting Dose in Adult Healthy Volunteer. Rockville, MD: US Food and Drug Administration.

Garsetti, M., Pellegrini, N., Baggio, C. & Brighenti, F. 2000. Antioxidant capacity in human faeces. British Journal of Nutrition 84: 705-710.

Golli, E.N. 2016. Toxicity Induced after subchronic administration of the synthetic food dye tartrazine in adult rats, role of oxidative stress. Recent Advance in Biological Medicine 2: 21-28.

Goth, L. 1991. A simple method for determination of serum catalase and reversion of reference range. Clinica Chimica Acta 2(3): 143-151.

Grootveld, M. & Halliwell, B. 1986. Aromatic hydroxylation as a potential measure of hydroxyl radical formation in vivo. Biochemical Journal 237: 449-504.

Halliwell, B. 1992. Reactive oxygen species and the central nervous system. Journal of Neurochemistry 59: 1609-1623

Halliwell, B., Zentella, A., Gomez, E.O. & Kershenobich, D. 1997. Antioxidants and human disease: A general introduction. Nutrition Reviews 55(1): 44.

Hassan, G. 2010. Effects of some synthetic coloring additives on DNA damage and chromosomal aberrations of rats. Arab Journal of Biotechnology 13(1): 13-24.

Himri, I., Bellahcen, S., Souna, F., Belmekki, F., Aziz, M., Bnouham, M., Zoheir, J., Berkia, Z., Mekhfi, H. & Saalaoui, E. 2011. A 90-day oral toxicity study of Tartrazine, a synthetic food dye, in Wistarrats. International Journal of Pharmacy and Pharmacological Science 3: 159-169.

Juretic, D., Motejlkova, A., Kunovic, B., Rekic, B., Mestric, Z.F., Vujic, L., Mesic, R., Bajalo, J.L. & Rudolf, V.S. 2006. Paraoxanase/arylesterase in serum of patients with type-II diabetes mellitus. Acta Pharmaceutical 56: 59-68.

Kalpana, C. & Menon, V.P. 2004. Modulatory effects of Curcumin on lipid peroxidation and antioxidant status during nicotine-induced toxicity. Polish Journal of Pharmacology 56: 581-586.

Kaur, M. & Sandhir, R. 2006. Comparative effects of acute and chronic carbofuran exposure on oxidative stress and drug-metabolizing enzymes in liver. Drug Chemistry and Toxicology 29(4): 415-421.

Khadija, A.A., Ati, S., Mohammed, A.M. & Mohamed, H.E. 2009. Response of broiler chicks to dietary monosodium glutamate. Pakistan Veterinary Journal 29(4): 165-168.

Kharwar, R.K. & Haldar, C. 2012. Daily variation in antioxidant enzymes and lipid peroxidation in lungs of a tropical bird Perdiculaasiatica: Role of melatonin and nuclear receptor RORα. Comparative Biochemistry and Physiology Part A. Molecular and Integrative Physiology 162: 296-302.

Kim, J.H., Hahm, D.H., Yang, D.C., Lee, H.J. & Shim, I. 2005. Effect of crude saponin of Korean red ginseng on high-fat diet-induced obesity in the rat. Journal of Pharmacological Science 97: 124-131.

Liu, T., Chen, C., Chen, C. & Chi, C. 1999. Safrole-induced oxidative damage in the liver of Sprague-Dawley rats. Food and Chemical Toxicology 37(7): 697-702.

Mehedi, N., Ainad-Tabet, S., Mokrane, N., Addou, S., Zaoui, C., Kheroua, O. & Saidi, D. 2009. Reproductive toxicology of tartrazine (FD and C Yellow No. 5) in swiss albino mice. American Journal of Pharmacology and Toxicology 4(4): 130-135.

Mourad, I.M. & Noor, N.A. 2011. Aspartame (a widely used artificial sweetener) and oxidative stress in the rat cerebral cortex. International Journal of Biomedicine Science 2(1): 4-10.

Moutinho, I.L.D., Bertges, L.C. & Assis, R.V.C. 2007. Prolonged use of the food dye tartrazine (FD&C yellow no 5) and its effects on the gastric mucosa of Wistar rats. Brazilian Journal of Biology 67(1): 141-145.

Mpountoukas, P., Pantazaki, A., Kostareli, E., Christodoulou, P., Kareli, D., Poliliou, S. & Lialiaris, T. 2010. Cytogenetic evaluation and DNA interaction studies of the food colorants amaranth, erythrosine and tartrazine. Food and Chemical Toxicology 48(10): 2934-2944.

Murray, R. 1996. Metabolism of xenobiotics. In Harper's Biochemistry. Stamford: Appleton & Lange. p. 868.

Orozco, M.N., Solomons, N.W., Schumann, K., Friel, J.K. & de Montenegro, A.L.M. 2010. Antioxidant-rich oral supplements attenuate the effects of oral iron on in situ oxidation susceptibility of human feces. Journal of Nutrition 140(6): 1105-1110.

Owen, J.B. & Butterfield, D.A. 2010. Measurement of oxidized/reduced glutathione ratio. In Protein Misfolding and Cellular Stress in Disease and Aging: Concept, Protocols, edited by Bross, P. & Gregersan, N. New York: Humana Press. pp. 269-277.

Pincemail, J., Bonjean, K., Cayeux, K. & Defraigne, J.O. 2002. Me canismes physiologiques de la defense antioxydante. Nutrition and Clinical Metabolism 16: 233-239.

Rahal, A., Kumar, A., Singh, V., Yadav, B., Tiwari, R., Chakraborty, S. & Dhama, K. 2014. Oxidative stress, prooxidants, and antioxidants: The interplay. BioMed Research International 2014. Article ID. 761264.

Rajesh, H., Rao, S.N., Megha Rani, N., Prathima, K., Shetty, R.E.P. & Chandrashekhar, R. 2013. Phytochemical analysis of methanolic extract of Curcuma longa Linn. International Journal of Universal Pharmacy and Bio Sciences 2(2): 39-45.

Rao, P., Bhat, R.P., Sudershan, R.V. & Krishna, T.P. 2006. Consumption of synthetic food colours during festivals in Hyderabad, India. British Food Journal 107: 276-284.

Rutkowski, M., Grzegorczyk, K. & Paradowski, M.T. 2005. Colorimetric method of blood plasma total vitamin E determination - the own modification of Tsen method. Diagnostic Laboratory 41: 375.

Rutkowski, M., Grzegorczyk, K., Gendek, E. & Kedziora, J. 2006. Laboratory convenient modification of Bessey method for vitamin A determination in blood plasma. Journal of Physiology and Pharmacology 57: 221.

Rutkowski, M. & Grzegorczyk, K. 1998. Kolorymetryczne oznaczanie stezenia witaminy C osoczu krwi przy uzyciu odczynnika fosforowolframianowego - modyfikacja metody Kyawa [Colorimetric determination of vitamin C concentration in blood plasma with phosphotungstate reagent- a modification of Kyaw method]. Diagnostic Laboratory 4: 243 [in Polish].

Sahu, S.C. 2002. Dual role of organosulfur compounds in foods. Journal of Environmental Science and Health Part C Environmental Carcinogen and Ecotoxicology Reviews 20(1): 61-76.

Saima, S. 2011. Effect of artificial food coloring agents on health biomarkers, antioxidant enzymes, hepatic functions, metabolic hormones and DNA dam