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Bioactivity evaluation and HPLC UV-VIS based quantification of antioxidant secondary metabolites from extract and fractions of Bistorta amplexicaulis rhizome
Corresponding Author(s) : Muhammad Sheeraz Ahmad
Cellular and Molecular Biology,
Vol. 65 No. 1: Issue 1
Abstract
Bistorta amplexicaulis is a popular medicinal plant and reported as rich source of antioxidant compounds. The present study was designed for antioxidant and anticancer potential of polarity based fractions of B. amplexicaulis and its correlation to the secondary metabolites quantified by HPLC-UV/VIS.Crude extract was prepared by maceration method and polarity based fractions were prepared by solvent-solvent extraction. Antioxidant and anticancer potential was investigated by using various physiological and non-physiological assays while secondary metabolites rutin, naringin and quercetin present in extract and fractions were quantified by using HPLC- UV/VIS. All extracts showed Antioxidant potential but highest activity was obtained with ethyl acetate fraction (DPPH IC50 5.76±0.03 µg/ml, ABTS IC50 0.74±0.1 µg/ml, Total Antioxidant Assay 72.55±0.098 Ascorbic acid equivalents, Super oxide radical scavenging assay IC506.86±0.1909 µg/ml, Hydroxyl radical scavenging assay IC50 0.96±0.1690 µg/ml). The cytotoxicity of fractions against HepG2 cell lines showed lowest ell viability in n-hexane fraction (11%). The results revealed that ethyl acetate fraction of B. amplexicaulis can be a potential source of novel antioxidant compounds while n hexane fraction could provide anticancer compounds. A new method of simultaneous quantification of three flavonoids by using UV/VIS detector is reported in this study.
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- Cai J, Xie S, Feng J, Wang F P, Xu Q F. Protective Effect of Polygonum orientale L. Extracts against Clavibatermichiganense subsp. sepedonicum, the Causal Agent of Bacterial Ring Rot of Potato. PLoS One 2013; 8, 68480.
- Widodo N, Priyandoko D, Shah N, Wadhwa R, Kaul S C. Selective killing of cancer cells by Ashwagandha leaf extract and its component Withanone involves ROS signaling. PLoS One 2010; 5(10), e13536.
- Kurata N, Moore G, Nagamura Y, Foote T, Yano M, Minobe Y, Gale M. Conservation of genomic structure between rice and wheat. Nature Biotechnol 1994; 12, 276-278.
- Prior R L. Fruit and vegetables in the prevention of cellular oxidative damage. AJCN 2003; 78, 570S-578S.
- Pietta P G. Flavonoids as antioxidants. J NAT PROD 2000; 63, 1035-1042.
- Turkmen N, Sari F, Velioglu Y S. Effect of extraction solvents on concentration and antioxidant activity of black and black mate polyphenols determined by ferrous tartrate and Folin-Ciocalteu methods. Food Chem 2006; 99, 838841.
- Zheng W, Wang S Y. Antioxidant activity and phenolic compounds in selected herbs. Journal of Agriculture and Food Chemistry 2001; 49, 5165–5170.
- Pan Y X, Wang X. Cytotoxic withanolides from the flowers of Daturametel. J NAT PROD 2007; 70, 1127-1132.
- Zaman R, Akthar M S, Khan M S. In vitro antibacterial screening of Anethumgraveolen L. fruit Cichoriumintybus L. leaf, Plantagoovata L. seed husk and Polygonumviviparum L. root extracts against Helibacterpylori. Int. J. Pharm 2006; 2(6), 674-677.
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- Xiang M X, Xu Z, Su H W, Hu J, Yan Y J. Emodin-8-O-β-D-Glucoside from Polygonumamplexicaule D. Don var. SinenseForb. Promotes proliferation anddifferentiation of osteoblastic MC3T3-E1 cells. Molecules 2011; 16, 728-737.
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- Miliauskas G, Venskutonis PR, and Beek TA. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chem 2004; 85: 231-237.
- Lim Y Y, Lim T T, Tee J J. Antioxidant properties of several tropical fruits: A comparative study. Food Chem 2007; 103, 1003-1008.
- Oyaizu M. Studies on product of browning reaction prepared from glucose amine. Japanese Journal of Nutrition and dietetics, 1986, 44, 307-315.
- Halliwell B, Gutteridge J M C. Free radicals in biology and medicine. Oxford University Press; 1999.
- Obeid H K, Allen M S, Bedgood D R, Prenzler P D, Robards K. Investigation of Australian olive mill waste for recovery of biophenols. J Agric Food Chem 2005 53, 9911-9920.
- Re R, Pellegrini N, Proteggente A, Pannala A, Yang M. Rice-Evans C Antioxidant activity applying an improved ABTS radical cationdecolorization assay. Free Radical Biological and Medicine. 1999, 26, 1231-1237.
- Prieto P, Pineda M, Anguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a Phosphomolybdenum Complex: Specific application to the determination of Vitamin E. Anal. Biochem 1999; 269, 337-341.
- Beauchamp C O, Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal. Biochem 1971; 44: 276-287.
- Stalikas C D. Micelle-mediated extraction as a tool for separation and preconcentration in metal analysis. Trends Anal. Chem 2002; 21, 343– 355.
- Skehan P, Storeng H, Scudiero D, Monks A, Mcmahon J, Vistica D, Warren J T, Bokesch H, Kenney S, Boyd M R. New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 1990; 82, 1107-1112.
- Dai J, Russell J M. Plant Phenolics: Extraction, Analysis and Their Antioxidant and Anticancer Properties. Molecules 2010; 15(10), 7313-52.
- Brand-Williams W, Cuvelier M E, Berset C. Use of a free radical method to evaluate antioxidant activity. Food Sci. Technol 1995; 28: 25-30.
- Nabavi S M, Ebrahimzadeh M A, Nabavi S F, Hamidinia S F, Bekhradnia A R. Determination of antioxidant activity, phenol and flavonoids content of Parrotia persica Mey. Pharmacologyonline 2008a; 2: 560-567.
References
Cai J, Xie S, Feng J, Wang F P, Xu Q F. Protective Effect of Polygonum orientale L. Extracts against Clavibatermichiganense subsp. sepedonicum, the Causal Agent of Bacterial Ring Rot of Potato. PLoS One 2013; 8, 68480.
Widodo N, Priyandoko D, Shah N, Wadhwa R, Kaul S C. Selective killing of cancer cells by Ashwagandha leaf extract and its component Withanone involves ROS signaling. PLoS One 2010; 5(10), e13536.
Kurata N, Moore G, Nagamura Y, Foote T, Yano M, Minobe Y, Gale M. Conservation of genomic structure between rice and wheat. Nature Biotechnol 1994; 12, 276-278.
Prior R L. Fruit and vegetables in the prevention of cellular oxidative damage. AJCN 2003; 78, 570S-578S.
Pietta P G. Flavonoids as antioxidants. J NAT PROD 2000; 63, 1035-1042.
Turkmen N, Sari F, Velioglu Y S. Effect of extraction solvents on concentration and antioxidant activity of black and black mate polyphenols determined by ferrous tartrate and Folin-Ciocalteu methods. Food Chem 2006; 99, 838841.
Zheng W, Wang S Y. Antioxidant activity and phenolic compounds in selected herbs. Journal of Agriculture and Food Chemistry 2001; 49, 5165–5170.
Pan Y X, Wang X. Cytotoxic withanolides from the flowers of Daturametel. J NAT PROD 2007; 70, 1127-1132.
Zaman R, Akthar M S, Khan M S. In vitro antibacterial screening of Anethumgraveolen L. fruit Cichoriumintybus L. leaf, Plantagoovata L. seed husk and Polygonumviviparum L. root extracts against Helibacterpylori. Int. J. Pharm 2006; 2(6), 674-677.
Lia X, Matsumotob K, Murakamib Y, Tezukaa Y, Wud Y, Kadotaa S. Neuroprotective effects of Polygonummultiflorumon nigrostriatal dopaminergic degeneration induced by paraquat and maneb in mice. Pharmacol. Biochem. Behav 2005; 82(2), 345-352.
Matin A, Khan M A, Ashraf M, Qureshi R A. Traditional use of herbs, shrubs and trees of Shogran Valley, Mensehra, Pakistan. Pak J Biol Sci 2001; 4 (9), 1101-1107.
Sabeen M, Ahmad S S. Exploring the Folk Medicinal Flora of Abbotabad City, Pakistan. Ethnobot Leaflets 2009; 13, 810-33.
Xiang M X, Xu Z, Su H W, Hu J, Yan Y J. Emodin-8-O-β-D-Glucoside from Polygonumamplexicaule D. Don var. SinenseForb. Promotes proliferation anddifferentiation of osteoblastic MC3T3-E1 cells. Molecules 2011; 16, 728-737.
Batool S, Ullah S, Tabassum S, Bilal A, Faisal A, Saleem R S, Ahmad M S. Effect of Extraction Methodologies on Antioxidant Potential and Synergistic Anticancer Behavior of Leaf Extract of Polygonum amplexicaule Against HCT-116 Cells. Iran J Sci Technol Trans A Sci 2018; https://doi.org/10.1007/s40995-018-0532-x.
Miliauskas G, Venskutonis PR, and Beek TA. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chem 2004; 85: 231-237.
Lim Y Y, Lim T T, Tee J J. Antioxidant properties of several tropical fruits: A comparative study. Food Chem 2007; 103, 1003-1008.
Oyaizu M. Studies on product of browning reaction prepared from glucose amine. Japanese Journal of Nutrition and dietetics, 1986, 44, 307-315.
Halliwell B, Gutteridge J M C. Free radicals in biology and medicine. Oxford University Press; 1999.
Obeid H K, Allen M S, Bedgood D R, Prenzler P D, Robards K. Investigation of Australian olive mill waste for recovery of biophenols. J Agric Food Chem 2005 53, 9911-9920.
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M. Rice-Evans C Antioxidant activity applying an improved ABTS radical cationdecolorization assay. Free Radical Biological and Medicine. 1999, 26, 1231-1237.
Prieto P, Pineda M, Anguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a Phosphomolybdenum Complex: Specific application to the determination of Vitamin E. Anal. Biochem 1999; 269, 337-341.
Beauchamp C O, Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal. Biochem 1971; 44: 276-287.
Stalikas C D. Micelle-mediated extraction as a tool for separation and preconcentration in metal analysis. Trends Anal. Chem 2002; 21, 343– 355.
Skehan P, Storeng H, Scudiero D, Monks A, Mcmahon J, Vistica D, Warren J T, Bokesch H, Kenney S, Boyd M R. New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 1990; 82, 1107-1112.
Dai J, Russell J M. Plant Phenolics: Extraction, Analysis and Their Antioxidant and Anticancer Properties. Molecules 2010; 15(10), 7313-52.
Brand-Williams W, Cuvelier M E, Berset C. Use of a free radical method to evaluate antioxidant activity. Food Sci. Technol 1995; 28: 25-30.
Nabavi S M, Ebrahimzadeh M A, Nabavi S F, Hamidinia S F, Bekhradnia A R. Determination of antioxidant activity, phenol and flavonoids content of Parrotia persica Mey. Pharmacologyonline 2008a; 2: 560-567.