Antioxidant Activity of Pericarp Extract from Different Varieties of Pomegranate Fruit

Document Type : Original Research

Authors
1 Department of Biochemistry, Shiraz University, Shiraz, Islamic Republic of Iran.
2 Department of Surgery, School of Veterinary Medicine, Shiraz University, Shiraz, Islamic Republic of Iran.
Abstract
The pomegranate Punicagranatum fruit pericarp, contain polyphenolic compounds including alpha and beta punicalagins and ellagic acid, which exhibit remarkable antioxidant activities. The aim of this study was to purify and quantify the phenolic components from different varieties of Pomegranate Pericarp Extracts (PPEs) and determine their antioxidant properties. Methanolic and aqueous extracts of four pomegranate cultivars (Shahvar, Siahsorfeh, Torshsabz and Abdorahimkhany, from Shiraz, Iran) were prepared and total phenolic content of PPEs was determined. PPE components were further purified by XAD-16 column chromatography followed by LH-20 gel filtration. The eluted components were subjected to HPLC analysis to differentiate and quantify polyphenolic compounds. Antioxidant activity was measured using DPPH assay. The result revealed significant difference in total phenolic contents and phenolic components in four cultivars. Total phenolic content in methanolic PPE was significantly (P< 0.05) higher than aqueous extracts. Shahvar cultivar had the highest total phenols (11.72±0.01 mg mL-1 in water, 17.7±0.12 mg mL-1 in methanol) and exhibited the most antioxidant property among cultivars. Analysis of components of Shahvar PPE by HPLC showed that proportions of different phenolic components were alpha punicalagin (28.34±2.12%), beta punicalagin (39.75±2.14%) and ellagic acid (3.49±0.93%). Antioxidant activity of alpha punicalagin, beta punicalagin, punicalagin (sum of alpha+beta) and ellagic acid was 50.8, 60.5, 56, and 27%, respectively. Taken together, these results indicate that PPE of Shahvar cultivar is an excellent source of natural antioxidants and can potentially replace the current synthetic antioxidants in food and pharmaceutical industries.

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1. Aguilar ,C.N, Aguilera-Carbo, A., Robledo, A., Ventura, J., Belmares R., Martinez, D., Rodríguez-Herrera, R. and Contreras, J. 2008. Production of antioxidant nutraceuticals by solid-state cultures of pomegranate (Punica granatum) peel and creosote bush (Larrea tridentata) leaves. Food Technol. Biotech. 46: 218-222.
2. Aviram, M., Dornfeld, L., Rosenblat, M., Volk,ova, N., Kaplan, M., Coleman, R., Hayek, T., Presser, D. and Fuhrman, D. 2000. Pomegranate juice consumption reduces oxidative stress, atherogenic modifications to LDL, and platelet aggregation: studies in humans and in atherosclerotic apolipoprotein E–deficient mice. Am. J. Clin. Nutr. 71: 1062-1076.
3. Aviram ,M., Rosenblat, M., Gaitini, D., Nitecki, S., Hoffman, A., Dornfeld, L., Volkova, N., Presser, D., Attias, J., Liker, H.and Hayek, T. 2004. Pomegranate juice consumption for 3 years by patients with carotid artery stenosis reduces common carotid intima-media thickness, blood pressure and LDL oxidation. Clin. Nutr. 23: 423-433.
4. Banihani, S., Swedan, S. and Alguraan, Z. 2013. Pomegranate and type 2 diabetes. Nutr. Res. 33: 341-348.
5. Basu, A. and Penugonda, K. 2009. Pomegranate juice: a heart-healthy fruit juice. Nutr. Rev. 67: 49-56.
6. Celik, I., Temur, A. and Isik, I. 2009. Hepatoprotective role and antioxidant capacity of pomegranate (Punica granatum) flowers infusion against trichloroacetic acid-exposed in rats. Food Chem. Toxicol. 47: 145-149.
7. Chidambara Murthy, K.N., Jayaprakasha, G.K. and Singh R.P. 2002. Studies on antioxidant activity of pomegranate (Punica granatum) peel extract using in vivo models. J. Agric. food Chem. 50(17): p. 4791-4795.
8. Dahham, S.S., Ali, M. N., Tabassum, H. and Khan, M. 2010. Studies on antibacterial and antifungal activity of pomegranate (Punica granatum L.). Am. Eurasian J. Agric. Environ. Sci. 9: 273-281.
9. Dikmen, M., Ozturk, N. and, Ozturk, Y. 2011. The antioxidant potency of Punica granatum L. fruit peel reduces cell proliferation and induces apoptosis on breast cancer. J. Medic. Food, 14: 1638-1646.
10. Doig AJ, Williams DH, Oelrichs PB, Baczynskyj L (1990) Isolation and structure elucidation of punicalagin, a toxic hydrolysable tannin, from Terminalia oblongata. J Chem Soc Perkin Trans 1: 2317-2321.
11. Ferreira, D. 2007. Antioxidant, antimalarial and antimicrobial activities of tannin-rich fractions, ellagitannins and phenolic acids from Punica granatum L. L. Planta Med. 73: 461-467.
12. Gil, M.I., Tomás-Barberán, F.A., Hess-Pierce, B., Holcroft, D.M. and Kader, A.A. 2000. Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing. J. Agric. Food Chem. 48: 4581-4589.
13. Hayouni, E.A., Miled, K., Boubaker, S., Bellasfar, Z., Abedrabba, M., Iwaski, H., Oku, H., Matsui, T., Limam, F. and Hamdi, M. 2011. Hydroalcoholic extract based-ointment from Punica granatum L. peels with enhanced in vivo healing potential on dermal wounds. Phytomedicine 18: 976-984.
14. Hong, M.Y., Seeram, N.P. and Heber, D. 2008. Pomegranate polyphenols down-regulate expression of androgen-synthesizing genes in human prostate cancer cells overexpressing the androgen receptor. J. Nut. Biochem. 19: 848-855.
15. Jurenka, J.S. 2008 Therapeutic applications of pomegranate (Punica granatum L.): A review. Altern. Med. Rev. 13: 128-44.
16. Kanatt, S.R., Chander, R. and Sharma, A. 2010. Antioxidant and antimicrobial activity of pomegranate peel extract improves the shelf life of chicken products. Int. J. Food Sc.i Tech. 45: 216-222.
17. Kaplan M, Hayek T, Raz A, Coleman R, Dornfeld L, Vaya J, Aviram M (2001) Pomegranate juice supplementation to atherosclerotic mice reduces macrophage lipid peroxidation, cellular cholesterol accumulation and development of atherosclerosis. J Nutr 131: 2082-2089.
18. Kulkarni, A.P., Aradhya, S.M. and Divakar, S. 2004. Isolation and identification of a radical scavenging antioxidant–punicalagin from pith and carpellary membrane of pomegranate fruit. Food Chem. 87: 551-557.
19. Kwak, H.M., Young Jeon, S., Sohng, B.H., Kim, J.G., Lee, J.M., Lee, K.B., Jeong, H.H., Hur, J.M., Kang, Y.H. and Song, K.S. 2005. β-Secretase (BACE1) inhibitors from pomegranate (Punica granatum) husk. Arch. Pharm. Res. 28: 1328-1332.
20. Levin, G.M. 1994. Pomegranate (Punica granatum) plant genetic resources in Turkmenistan. Plant Genetic Resources Newsletter (IPGRI/FAO) , (no.97) p. 31-36.
21. Longtin, R. 2003. The pomegranate: nature’s power fruit? J. Natl. Cancer I. I5: 346-348.
22. Lu, J., Ding, K. and Yuan, Q. 2008. Determination of punicalagin isomers in pomegranate husk. Chromatographia 68: 303-306.
23. Mertens-Talcott, S.U. and Percival, S.S. 2005. Ellagic acid and quercetin interact synergistically with resveratrol in the induction of apoptosis and cause transient cell cycle arrest in human leukemia cells. Cancer Lett. 218: 141-151.
24. Middha, S.K., Usha, T. and RaviKiran, T. 2012. Influence of Punica granatum L. on region specific responses in rat brain during Alloxan-Induced diabetes. Asian Pac. J. Trop. Biomed. 2: S905-S909.
25. Okonog, S., Duangrat, C., Anuchpreeda, S., Tachakittirungrod, S. and Chowwanapoonpohn, S. 2007. Comparison of antioxidant capacities and cytotoxicities of certain fruit peels. Food Chem. 103: 839-846.
26. Ricci, D., Giamperi, L., Bucchini, A. and Fraternale, D. 2006. Antioxidant activity of Punica granatum fruits. Fitoterapia, 77: 310-312.
27. Rosenblat, M., Volkova, N. and Aviram, M. 2013. Pomegranate phytosterol (β-sitosterol) and polyphenolic antioxidant (punicalagin) addition to statin, significantly protected against macrophage foam cells formation. Atherosclerosis 226: 110-117.
28. Rouanet, J.M., Décordé, K., Del Rio, D., Auger, C., Borges, G., Cristol, J.P., Lean, M.E.J. and Crozier, A. 2010. Berry juices, teas, antioxidants and the prevention of atherosclerosis in hamsters. Food Chem. 118: 266-271.
29. Sadasivam, S. and Manickam, A. 1996. Phenolics. In: Biochemical Methods. New Age International. New Delhi. p 195.
30. Seeram, N.P., Lee, R., Heber, D. 2004. Bioavailability of ellagic acid in human plasma after consumption of ellagitannins from pomegranate (Punica granatum L.) juice. Clin. Chim. Acta, 348: 63-68.
31. Seeram, N.P., Adams, L.S., Henning, S.M., Niu, Y., Zhang, Y., Nair, M.G. and Heber, D. 2005a. In vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegranate tannin extract are enhanced in combination with other polyphenols as found in pomegranate juice. The Journal of nutritional biochemistry J. Nutr. Biochem. 16: 360-367.
32. Seeram, N., Lee, R., Hardy, M. and Heber, D.. 2005b. Rapid large scale purification of ellagitannins from pomegranate husk, a by-product of the commercial juice industry. Sep. Purif. Technol. 41: 49-55.
33. Shukla, M., Gupta, K., Rasheed, Z., Khan, K.A. and Haqqi, T.M. 2008. Consumption of hydrolyzable tannins-rich pomegranate extract suppresses inflammation and joint damage in rheumatoid arthritis. Nutrition 24: 733-743.
34. Singh, R.P., Chidambara Murthy, K.N., and Jayaprakasha, G.K. 2002. Studies on antioxidant activity of pomegranate (Punica granatum) peel extract using in vivo models. J. Agric. Food Chem. 50: 4791-4795.
35. Srivastava. A., Rao, L.J.M. and Shivanandappa, T. 2007. Isolation of ellagic acid from the aqueous extract of the roots of Decalepis hamiltonii: antioxidant activity and cytoprotective effect. Food Chem. 103: 224-233.
36. Viuda‐Martos, M., Fernández‐López, J. and Pérez‐Álvarez, J. 2010. Pomegranate and its many functional components as related to human health: a review. Comp. Rev. in Food Sci. Food Safety, 9: 635-654.
37. Vroegrijk, I.O., van Diepen, J.A., van den Berg, S., Westbroek, I., Keizer, H., Gambelli, L., Hontecillas, R., Bas.saganya-Riera, J., Zondag, G.C., Romijn, J.A., Havekes, L.M, and Voshol, P.J. 2011. Pomoegranate seed oil, a rich source of punicic acid, prevents diet-induced obesity and insulin resistance in mice. Food Chem. Toxicol. 49: 1426-1430.
38. Xu, X. , Li, H., Hou, X., Li, D., He, S., Wan, C., Yin, P., Liu, M., Liu, F. and Xu, J. 2015. Punicalagin induces Nrf2/HO-1 expression via upregulation of PI3K/AKT pathway and inhibits LPS-induced oxidative stress in RAW264. macrophages. Mediators in Inflamm. 2015: 380218.