DETERMINATION OF ANTIOXIDANT CAPACITY AND GALLIC ACID CONTENT IN ETHANOLIC EXTRACT OF PUNICA GRANATUM L. LEAF
Â Objective: The present research work was carried out to evaluate the antioxidant potential of ethanolic extract of Punica granatum leaf (EPGL) that belongs to the family of Punicaceae and determine its gallic acid (GA) content using chromatography method.
Methods: Six complementary test systems, namely, 1,1-diphenyl 2-picryl hydrazine (DPPH), hydrogen peroxide (H2O2), superoxide (SO), nitric oxide (NO), hydroxyl (OH) radical scavenging, and reducing power activities were analyzed for determining antioxidant activity of EPGL. The simple and novel chromatography techniques such as thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were used for the detection and quantification of GA in EPGL.
Results: IC50 values of EPGL were found to be 136 Î¼g/ml for DPPH, 88.5 Î¼g/ml for H2O2, 16.8 Î¼g/ml for SO, 96.5 Î¼g/ml for NO, and 143 Î¼g/ml for OH. The ascorbic acid (AA) and GA were used as standard compounds. The absorbance of EPGL in reducing power assay was found to be 0.18 at 100 Î¼g/ ml, while AA and GA absorbance was found to be 0.24 and 0.4 at the same concentration. The amount of GA in EPGL was found to be 1.189 mg/g.
Conclusion: These findings suggested that EPGL could be a potential source of natural antioxidant, and HPLC method used for the determination of GA is simple, precise, accurate, and suitable for routine analysis of GA in EPGL.
2. Niki E, Shimaski H, Mino H. Antioxidantism-Free Radical and Biological Defense. Tokyo: Gakkai Syuppn Centre; 1994.
3. Temple NJ. Antioxidants and disease: More questions than answers. Nutr Res 2000;20:449-9.
4. La Vecchia C, Altieri A, Tavani A. Vegetables, fruit, antioxidants and cancer: A review of Italian studies. Eur J Nutr 2001;40:261-7.
5. Marques GS, Leao WF, Lyra MA, Peixoto MS, Monteiro RP, Rolim LA, et al. Comparative evaluation of UV/VIS and HPLC analytical methodologies applied for quantification of flavonoids from leaves of Bauhinia forficata. Rev Bras Farmacogn 2012;23:51-7.
6. Zollner T, Schwarz M. Herbal reference standards: Applications, definitions and regulatory requirements. Rev Bras Farmacogn 2013;23:1-21.
7. Altuner EM. Investigation of antimicrobial activity of Punica granatum L. fruit peel ash used for protection against skin infections as folk remedies especially after male circumcision. Afr J Microbiol Res 2011;5:3339-2.
8. Arun N, Singh DP. Punica granatum: A review on pharmacological and therapeutic properties. Int J Pharm Sci Res 2012;3:1240-5.
9. Choi JG, Kang OH, Lee YS, Chae HS, Oh YC, Brice OO, et al. In vitro and in vivo antibacterial activity of Punica granatum peel ethanol extract against salmonella. Evid Based Complement Alternat Med 2009;2011:1-8.
10. Dahham SS, Ali MN, Tabassum H, Khan M. Studies on antibacterial and antifungal activity of pomegranate (Punica granatum L.). Am Eur J Agric Environ Sci 2010;9:273-1.
11. Inabo HI, Fathuddin MM. In vivo antitrypanosomal potentials of ethyl acetate leaf extracts of Punica granatum against Trypanosoma brucei brucei. Adv Agr Biol 2011;1:82-8.
12. Moussa AM, Emam AM, Diab YM, Mahmoud ME, Mahmoud AS. Evaluation of antioxidant potential of 124 Egyptian plants with emphasis on the action of Punica granatum leaf extract on rats. Int Food Res J 2011;18:535-2.
13. Sreedevi P, Vijayalakshmi K, Venkataeswari R. Phytochemical evaluation of Punica granatum L. Leaf extract. Int J Curr Pharm Res 2017;9:14-8.
14. Singh RP, Murthy KN, Jayaprakasha GK. Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extracts using in vitro models. J Agric Food Chem 2002;50:81-6.
15. Ebrahimzadeh MA, Nabavi SF, Nabavi SM. Antioxidant activity of leaves and inflorescence of Eryngium caucasicum Trautv at flowering stage. Phcog Res 2009;1:435-9.
16. Liu F, Ooi VE, Chang ST. Free radical scavenging activites of mushroom polysaccharide extracts. Life Sci 1997;60:763-1.
17. Govindharajan R, Rastogi S, Vijayakumar M, Rawat AK, Shirwaikar A, Mehrotra S, et al. Studies on the antioxidant activities of Desmodium gangeticum. Biol Pharm Bull 2003;26:1424-7.
18. Halliwell B, Gutteridge JM, Cross CE. Free radicals, antioxidants, and human disease: Where are we now? J Lab Clin Med 1992;119:598-20.
19. Oyaizu M. Studies on products of browning reaction-antioxdative activities of products of browning reaction prepared from glucoseamine. Jpn J Nutr Diet 1986;44:307-5.
20. Marston A, Kissling J, Hostettmann K. A rapid TLC bioautographic method for the detection of acetylcholineesterase and butyrylcholineesterase inhibitors in plants. Phytochem Anal 2002;13:51-4.
21. Kardani K, Gurav N, Solanki B, Patel P, Patel B. RP-HPLC method development and validation of gallic acid in polyherbal tablet formulation. J Appl Pharm Sci 2013;3:37-2.
22. Halliwell B, Gutteridge JM. Free Radicals in Biology and Medicine. UK: Oxford University Press; 1998.
23. Yazdanparast R, Ardestani A. In vitro antioxidant and free radical scavenging activity of Cyperus rotundus. J Med Food 2007;10:667-4.
24. Schimoi K, Masuda S, Shen B, Furugori M, Kinae N. Radioprotective effects of antioxidative plant flavonoids in mice. Mutat Res 1996;350:153-61.
25. Sakat S, Juvekar AR, Gambhire MN. In-vitro antioxidant and anti-inflammatory activity of methanol extract of Oxalis corniculata Linn. Int J Pharm Pharm Sci 2010;2:146-5.
26. Zhang LH, Li LL, Li YX, Zhang YH. In vitro antioxidant activities of fruits and leaves of Pomegranate. Acta Hortic 2008;765:31-4.
27. Gheith I, El-Mahmoudy A. Potent anti-oxidant and anti-inflammatory potentials of Punica granatum leaf and flower hydromethanolic extracts In vitro. Biosci J 2017;33:434-46.
28. Soares JR, Dins TC, Cunha AP, Almeida LM. Antioxidant activities of some extracts of Thymus Zygis. Free Radic Res 1997;26:469-8.
29. Vinodhini S, Shri Preethi M, Nusrath Fathima N, Kushwaha SS, Devi Rajeswari V. Antioxidant and free radical scavenging activity of Punica granatum leaf extracts. Asian J Pharm Clin Res 2016;9:17-20.
30. MacDonald-Wicks LK, Wood LG, Garg ML. Methodology for the determination of biological antioxidant capacity in vitro: A review. J Sci Food Agric 2006;86:2046-6.
31. Rummun N, Somanah J, Ramasaha S, Bahorun T, Neergheen-Bhujan SN. Bioactivity of nonedible parts of Punica granatum L.: A potential source of functional ingredients. Int J Food Sci 2013;2013:1-12.
32. Parejo I, Viladomat F, Bastida J, Rosas-Romero A, Flerlage N, Burillo J. et al. Comparison between the radical scavenging activity and antioxidant activity of six distilled and nondistilled mediterranean herbs and aromatic plants. J Agri Food Chem 2005;50:6882-90.
33. Yen G, Duh P. Scavenging effect of methanolic extracts of peanut hulls on free-radical and active-oxygen species. J Agri Food Chem 1994;42:629-2.
34. Hossain H, Ahmed T, Howlader MS, Dey SK, Hira A, Ahmed A, et al. In-vitro antioxidant potential from the leaves of Punica granatum Linn grown in Bangladesh. Int J Phytopharmcol Res 2012;2:160-6.
35. Huie RE, Padmaja S. The reaction of no with superoxide. Free Radic Res Commun 1993;18:195-9.
36. Babu BH, Shylesh BS, Padikkala J. Antioxidant and hepatoprotective effect of Acanthus ilicifolius. Fitoterapia 2001;72:272-7.
37. Ravishankar N, Sivaraj C, Seeni S, Joseph J, Raaman N. Antioxidant activities and phytochemical analysis of methanol extract of leaves of Hypericum Hookerianum. Int J Pharm Pharm Sci 2014;6:456-60.
38. Nabavi SM, Ebrahimzadeh MA, Nabavi SF, Fazelian M, Eslami B. In vitro antioxidant and free radical scavenging activity of Diospyros lotus and Pyrus boissieriana growing in Iran. Pharmacogn Mag 2009;5:122- 6.
39. Tanaka M, Kuei CW, Nagashima Y, Taguchi T. Application of antioxidative Maillard reaction products from histidine and glucose to sardine products. Nippon Suisan Gakkaishi 1988;54:1409-4.
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