Please wait a minute...
Journal of Integrative Agriculture  2017, Vol. 16 Issue (08): 1808-1818    DOI: 10.1016/S2095-3119(16)61560-5
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
In vitro and in vivo antioxidant activities of three major polyphenolic compounds in pomegranate peel: Ellagic acid, punicalin, and punicalagin
SUN Yu-qing1, TAO Xin2, MEN Xiao-ming2, XU Zi-wei1, 2, WANG Tian1
1 College of Animal Science & Technology, Nanjing Agricultural University, Nanjing 210095, P.R.China
2 Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, P.R.China
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
Abstract      Pomegranates is abundant in polyphenols and is well-known for its antioxidant activity. Punicalagin (PG) is a major polyphenolic compound in the pomegranate peel. In certain conditions, PG can be hydrolyzed to punicallin (PL) and ellagic acid (EA), and PL can be further hydrolyzed to EA. PG, PL, and EA all play important roles in the antioxidant activity of pomegranate peels. This study was conducted to compare the in vitro antioxidant activity and in vivo anti-oxidative stress effects of PG, PL, and EA. For the in vitro test, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH·) and superoxide anion (O2-.) scavenging capacities, ferric-reducing antioxidant power (FRAP), and lipid peroxidation (LPO) inhibition capacities of PG, PL, and EA were tested. For the in vivo test, oxidatively stressed mice, which were induced by oxidized fish oil, were administrated PG, PL or EA (10 mg kg–1 d –1) for 21 days. The results showed that the in vitro antioxidant activity trends were EA>PG>PL>Trolox in scavenging DPPH?, PG>PL>EA≈Trolox in scavenging O2-. , EA>PG≈PL>Trolox in FRAP, and Trolox>PG>EA>PL in LPO inhibition. In the in vivo test, the EA treatment increased the average daily weight gain and total antioxidant capacity (T-AOC) in the plasma (P<0.05), liver (P<0.05), and intestine (P<0.05) in oxidatively stressed mice. It increased the superoxide dismutase (SOD) activity in the liver (P<0.05) and intestine (P<0.05). It increased the glutathione peroxidase (GSH-Px) activity in the intestine (P<0.05) and the intestinal villus height to crypt depth ratio (P<0.05). EA treatment decreased the malondialdehyde (MDA) content in the plasma (P<0.05), liver (P<0.05), and intestine (P<0.05) and the mRNA expressions of the pro-inflammatory factors, TNF-α (P<0.05), IFN-γ (P<0.05) and IL-6 (P<0.05). PL increased the SOD (P<0.05) and GSH-Px activities (P<0.05) in the intestine and decreased the MDA content (P<0.05) and the mRNA expressions of TNF-α (P<0.05) and IL-6 (P<0.05) in the intestine. PG increased the SOD activity (P<0.05) and GSH-Px activity (P<0.05) in the intestine and decreased the MDA content in the intestine (P<0.05) and IL-6 mRNA expression in the intestine (P<0.05). In summary, EA, PL, and PG all had powerful in vitro antioxidant capacities, and they had different antioxidant advantages in acting against different types of radicals; EA was more effective than PL and PG in protecting against oxidative injury in vivo, especially for intestinal injury. These findings suggest that multiple polyphenol compounds in pomegranate peel may exert superior antioxidant activity than single purified polyphenols; when using pomegranate peels as health-promoting additive in animal feed, raising EA content by methods of hydrolysis or fermentation in advance could achieve better effects.
Keywords:  ellagic acid        punicalin        punicalagin        antioxidant        oxidative stress  
Received: 02 August 2016   Accepted: 02 August 2017

This work was supported by the Science and Technology Department of Zhejiang Province, China (2012C12906-4), the Modern Agro-industry Technology Research System of China (CARS-36), the National Key Technology R&D Program, China (2012BAD39B03-4), and Agro-scientific Research in the Public Interest, China (201403047).

Corresponding Authors:  Correspondence XU Zi-wei, Tel: +86-571-86404398, Fax: +86-571-86400386, E-mail:   
About author:  SUN Yu-qing, Tel: +86-571-86404209, E-mail: sunyuqing1216;

Cite this article: 

SUN Yu-qing, TAO Xin, MEN Xiao-ming, XU Zi-wei, WANG Tian . 2017. In vitro and in vivo antioxidant activities of three major polyphenolic compounds in pomegranate peel: Ellagic acid, punicalin, and punicalagin. Journal of Integrative Agriculture, 16(08): 1808-1818.

Aluko R E, Monu E. 2003. Functional and bioactive properties of quinoa seed protein hydrolysates. Journal of Food Science, 68, 1254–1258.

Aqil F, Vadhanam M V, Gupta R C. 2012. Enhanced activity of punicalagin delivered via polymeric implants against benzo[a]pyrene-induced DNA adducts. Mutation Research, 743, 59–66.

Azofeifa G, Quesada S, Pérez A M, Vaillant F, Michel A. 2015. Pasteurization of blackberry juice preserves polyphenol-dependent inhibition for lipid peroxidation and intracellular radicals. Journal of Food Composition and Analysis, 42, 56–62.

Bajpai V K, Sharma1 A, Kang S C, Baek K H. 2014. Antioxidant, lipid peroxidation inhibition and free radical scavenging efficacy of a diterpenoid compound sugiol isolated from Metasequoia glyptostroboides. Asian Pacific Journal of Tropical Medicine, 7, 9–15.

Budilarto E S, Kamal-Eldin A. 2015. The supramolecular chemistry of lipid oxidation and antioxidation in bulk oils. European Journal of Lipid Science and Technology, 117, 1095–1137.

Cerdá B, Llorach R, Cerón J J, Espín J C, Tomás-Barberán F A. 2003. Evaluation of the bioavailability and metabolism in the rat of punicalagin, an antioxidant polyphenol from pomegranate juice. European Journal of Nutrition, 42, 18–28.

Davies K J. 1995. Oxidative stress: The paradox of aerobic life. Biochemical Society Symposium, 61, 1–31.

Gao J, Koshio S, Ishikawa M, Yokoyama S, Ren T, Komilus C F, Han Y. 2012. Effects of dietary palm oil supplements with oxidized and non-oxidized fish oil on growth performances and fatty acid compositions of juvenile Japanese sea bass, Lateolabrax Japonicus. Aquaculture, 324–325, 97–103.

Gulcin I. 2012. Antioxidant activity of food constituents: An overview. Archives of Toxicology, 86, 345–391.

Halliwell B, Whiteman M. 2004. Measuring reactive species and oxidative damage in vivo and in cell culture: How should you do it and what do the results mean? British Journal of Pharmacology, 142, 231–255.

de Heuvel E, Wallace L, Sharkey K A, Sigalet D L. 2012. Glucagon-like peptide 2 induces vasoactive intestinal polypeptide expression in enteric neurons via phophatidylinositol 3-kinase-γ signaling. American Journal of Physiology Endocrinology & Metabolism, 303, E994–E1005.

Hwang D F, Hour J L, Chen H M. 2000. Effect of taurine on toxicity of oxidized fish oil in rats. Food and Chemical Toxicology, 38, 585–591.

Ichinose T, Nobuyuki S, Takano H, Abe M. 2004. Liver carcinogenesis and formation of 8-hydroxy-deoxyguanosine in C3H/HeN mice by oxidized dietary oils containing carcinogenic dicarbonyl compounds. Food and Chemical Toxicology, 42, 1795–1803.

Landete J M. 2011. Ellagitannins, ellagic acid and their derived metabolites: A review about source, metabolism, functions and health. Food Research International, 44, 1150–1160.

Lansky E P, Newman R A. 2007. Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. Journal of Ethnopharmacology, 109, 177–206.

Larrosa M, Tomas-Barberan F A, Espin J C. 2006. The dietary hydrolysable tannin punicalagin releases ellagic acid that induces apoptosis in human colon adenocarcinoma

Caco-2 cells by using the mitochondrial pathway. Journal of Nutritional Biochemistery, 17, 611–625.

Li J K, Li G X, Zhao Y H, Yu C Z. 2009. Composition of pomegranate peel polyphenols and its antioxidant activities. Scientia Agricultura Sinica, 42, 4035–4041. (in Chinese).

Liang J, Li J K, Zhao W, Liu Y L. 2012. Effect of pomegranate peel polyphenols on lipid peroxidation in vitro. Journal of Food Science and Biotechnology, 31, 159–165. (in Chinese)

Liu R H. 2013. Health-promoting components of fruits and vegetables in the diet. Advances in Nutrition, 4, 384S–392S.

Martins S, Mercado D, Mata-Gómez M, Rodriguez L, Aguilera-Carbo A, Rodriguez R, Aguilar C N. 2009. Microbial production of potent phenolic-antioxidants through solid state fermentation. In: Singh O V, Harvey S P, eds., Sustainable Biotechnology. Springer, Netherlands. pp. 229–246.

Niwano Y, Saito K, Yoshizaki F, Kohno M, Ozawa T. 2011. Extensive screening for herbal extracts with potent antioxidant properties. Journal of Clinical Biochemistry and Nutrition, 48, 78–84.

Ogino H, Sakazaki F, Okuno T, Arakawa T, Ueno H. 2015. Oxidized dietary oils enhance immediate- and/or delayed-type allergic reactions in BALB/c mice. Allergology International, 64, 66–72.

Pérez-Jiménez J, Neveu V, Vos F, Scalbert A. 2010. Identification of the 100 richest dietary sources of polyphenols: An application of the phenol-explorer database. European Journal of Clinical Nutrition, 64, S112–S120.

Pfundstein B, El Desouky S K , Hull W E , Haubner R , Erben G, Owen R W. 2010. Polyphenolic compounds in the fruits of Egyptian medicinal plants (Terminalia ballerica, Terminalia chebula and Terminalia horrida): Characterisation, quantitation and determination of antioxidant capacities. Phytochemistry, 71, 1132–1148.

Poljsak B, Milisav I. 2012. The neglected significance of “antioxidative stress”. Oxidative Medicine and Cellular Longevity, 2012, 480895.

Qi K K, Wu J, Xu Z W. 2014. Effects of polyethylene glycosylation porcine glucagon-like peptide-2 on gene expression of tight junction proteins and inflammatory cytokines in a murine model of experimental colitis. Chinese Journal of Animal & Veterinary Sciences, 45, 1011–1017. (in Chinese)

Qiao Y, Sun J, Ding Y Y, Le G W, Shi Y H. 2013. Alterations of the gut microbiota in high-fat diet mice is strongly linked to oxidative stress. Applied Microbiology Biotechnology, 97, 1689–1697.

Ringseis R, Piwek N, Eder K. 2007. Oxidized fat induces oxidative stress but has no effect on NF-κB-mediated proinflammatory gene transcription in porcine intestinal epithelial cells. Inflammation Research, 56, 118–125.

Del Rio D, Costa L G, Lean M E, Crozier A. 2010. Polyphenols and health: What compounds are involved? Nutrition Metabolism and Cardiovascular Diseases, 20, 1–6.

Robledo A, Aguilera-Carbó A, Rodriguez R, Martinez J L, Garza Y, Aguilar C N. 2008. Ellagic acid production by Aspergillus niger in solid state fermentation of pomegranate residues. Journal of Industrial Microbiology & Biotechnology, 35, 507–518.

Rosillo M A, Sánchez-Hidalgo M, Cárdeno A, de la Lastra C A. 2011. Protective effect of ellagic acid, a natural polyphenolic compound, in a murine model of Crohn’s disease. Biochemical Pharmacology, 82, 737–745.

Rosillo M A, Sánchez-Hidalgo M, Cárdeno A, Aparicio-Soto M, Sánchez-Fidalgo S, Villegas I, de la Lastra C A. 2012. Dietary supplementation of an ellagic acid-enriched pomegranate extract attenuates chronic colonic inflammation in rats. Pharmacological Research, 66, 235–242.

Seeram N P, Adams L S, Henning S M, Niu Y, Zhang Y, Nair M G, Heber D. 2005. 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, 16, 360–367.

Seeram N P, Aronson W J, Zhang Y, Henning S M, Moro A, Lee R P, Sartippour M, Harris D M, Rettig M, Suchard M A, Pantuck A J, Belldegrun A, D Heber. 2007. Pomegranate ellagitannin-derived metabolites inhibit prostate cancer growth and localize to the mouse prostate gland. Journal of Agricultural and Food Chemistry, 55, 7731–7737.

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. Clinica Chimica Acta, 348, 63–68.

Sepúlveda L, Aguilera-Carbó A, Ascacio-Valdés J A, Rodríguez-Herrera R, Martínez-Hernández J L, Aguilar C N. 2012. Optimization of ellagic acid accumulation by Aspergillus niger GH1 in solid state culture using pomegranate shell powder as a support. Process Biochemistry, 47, 2199–2203.

Teel R W, Martin R M. 1988. Disposition of the plant phenol ellagic acid in the mouse following oral administration by gavage. Xenobiotica, 18, 397–405.

Wang Y, Zhang H, Liang H, Yuan Q. 2013. Purification, antioxidant activity and protein-precipitating capacity of punicalin from pomegranate husk. Food Chemistry, 138, 437–443.

Williams D J, Edwards D, Pun S, Chaliha M, Sultanbawa Y. 2014. Profiling ellagic acid content: The importance of form and ascorbic. Food Research International, 66, 100–106.

Xing R, Yu H H, Liu S, Zhang W W, Zhang Q B, Li Z E, Li P C. 2005. Antioxidant activity of differently regioselective chitosan sulfates in vitro. Bioorganic & Medicinal Chemistry, 13, 1387–1392.

Yeh Y H, Lee Y T, Hsieh H S, Hwang D F. 2010. Effect of red yeast rice on toxicity of oxidized cholesterol and oxidized fish oil in rats. e-SPEN, the European e-Journal of Clinical Nutrition and Metabolism, 5, e230–e237.
[1] Senouwa Segla Koffi DOSSOU, XU Fang-tao, Komivi DOSSA, ZHOU Rong, ZHAO Ying-zhong, WANG Lin-hai. Antioxidant lignans sesamin and sesamolin in sesame (Sesamum indicum L.): a comprehensive review and future prospects[J]. >Journal of Integrative Agriculture, 2023, 22(1): 14-30.
[2] WANG Pei-hong, WANG Sai, NIE Wen-han, WU Yan, Iftikhar AHMAD, Ayizekeranmu YIMING, HUANG Jin, CHEN Gong-you, ZHU Bo. A transferred regulator that contributes to Xanthomonas oryzae pv. oryzicola oxidative stress adaptation and virulence by regulating the expression of cytochrome bd oxidase genes[J]. >Journal of Integrative Agriculture, 2022, 21(6): 1673-1682.
[3] Muhammad Ahsan ASGHAR, JIANG Heng-ke, SHUI Zhao-wei, CAO Xi-yu, HUANG Xi-yu, Shakeel IMRAN, Bushra AHMAD, ZHANG Hao, YANG Yue-ning, SHANG Jing, YANG Hui, YU Liang, LIU Chun-yan, YANG Wen-yu, SUN Xin, DU Jun-bo. Interactive effect of shade and PEG-induced osmotic stress on physiological responses of soybean seedlings[J]. >Journal of Integrative Agriculture, 2021, 20(9): 2382-2394.
[4] WANG Yi-bo, HUANG Rui-dong, ZHOU Yu-fei. Effects of shading stress during the reproductive stages on photosynthetic physiology and yield characteristics of peanut (Arachis hypogaea Linn.)[J]. >Journal of Integrative Agriculture, 2021, 20(5): 1250-1265.
[5] Iram SHAFIQ, Sajad HUSSAIN, Muhammad Ali RAZA, Nasir IQBAL, Muhammad Ahsan ASGHAR, Ali RAZA, FAN Yuan-fang, Maryam MUMTAZ, Muhammad SHOAIB, Muhammad ANSAR, Abdul MANAF, YANG Wen-yu, YANG Feng. Crop photosynthetic response to light quality and light intensity[J]. >Journal of Integrative Agriculture, 2021, 20(1): 4-23.
[6] YE Yu-xiu, WEN Zhang-rong, YANG Huan, LU Wei-ping, LU Da-lei. Effects of post-silking water deficit on the leaf photosynthesis and senescence of waxy maize[J]. >Journal of Integrative Agriculture, 2020, 19(9): 2216-2228.
[7] WANG Yan, YAN Hao, WANG Qi, ZHENG Ran, XIA Kai, LIU Yang. Regulation of the phytotoxic response of Arabidopsis thaliana to the Fusarium mycotoxin deoxynivalenol[J]. >Journal of Integrative Agriculture, 2020, 19(3): 759-767.
[8] WEN Bing-xiao, Sajad Hussain, YANG Jia-yue, WANG Shan, ZHANG Yi, QIN Si-si, XU Mei, YANG Wen-yu, LIU Wei-guo. Rejuvenating soybean (Glycine max L.) growth and development through slight shading stress[J]. >Journal of Integrative Agriculture, 2020, 19(10): 2439-2450.
[9] Shahnawaz Ahmed, H S Rattanpal, Khalid Gul, Rouf Ahmad Dar, Akash Sharma. Chemical composition, antioxidant activity and GC-MS analysis of juice and peel oil of grapefruit varieties cultivated in India[J]. >Journal of Integrative Agriculture, 2019, 18(7): 1634-1642.
[10] ZHANG Bin, CUI Guo-bing, CHANG Chang-qing, WANG Yi-xu, ZHANG Hao-yang, CHEN Bao-shan, DENG Yi-zhen, JIANG Zi-de. The autophagy gene ATG8 affects morphogenesis and oxidative stress tolerance in Sporisorium scitamineum[J]. >Journal of Integrative Agriculture, 2019, 18(5): 1024-1034.
[11] LI Fu-xiang, LI Fu-hua, YANG Ya-xuan, YIN Ran, MING Jian. Comparison of phenolic profiles and antioxidant activities in skins and pulps of eleven grape cultivars (Vitis vinifera L.)[J]. >Journal of Integrative Agriculture, 2019, 18(5): 1148-1158.
[12] Bevly M. Mampholo, Martin M. Maboko, Puffy Soundy, Dharini Sivakumar. Variety-specific responses of lettuce grown in a gravel-film technique closed hydroponic system to N supply on yield, morphology, phytochemicals, mineral content and safety[J]. >Journal of Integrative Agriculture, 2018, 17(11): 2447-2457.
[13] ZHENG Yin-jian, ZHANG Yi-ting, LIU Hou-cheng, LI Ya-min, LIU Ying-liang, HAO Yan-wei, LEI Bing-fu. Supplemental blue light increases growth and quality of greenhouse pak choi depending on cultivar and supplemental light intensity[J]. >Journal of Integrative Agriculture, 2018, 17(10): 2245-2256.
[14] ZHOU Yan, DIAO Ming, CUI Jin-xia, CHEN Xian-jun, WEN Ze-lin, ZHANG Jian-wei, LIU Hui-ying. Exogenous GSH protects tomatoes against salt stress by modulating photosystem II efficiency, absorbed light allocation and H2O2- scavenging system in chloroplasts[J]. >Journal of Integrative Agriculture, 2018, 17(10): 2257-2272.
[15] PANG Yun-wei, JIANG Xiao-long, ZHAO Shan-jiang, HUANG Zi-qiang, ZHU Hua-bin. Beneficial role of melatonin in protecting mammalian gametes and embryos from oxidative damage[J]. >Journal of Integrative Agriculture, 2018, 17(10): 2320-2335.
No Suggested Reading articles found!