American Journal of Pharmacological Sciences
ISSN (Print): 2327-6711 ISSN (Online): 2327-672X Website: http://www.sciepub.com/journal/ajps Editor-in-chief: Srinivas NAMMI
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American Journal of Pharmacological Sciences. 2017, 5(1), 11-17
DOI: 10.12691/ajps-5-1-3
Open AccessArticle

In vitro Free Radical Scavenging Activities of the Saponin-rich Fractions from Vernonia amygdalina Del. (Compositae)

Chinaka Nwaehujor1, , Eridiong O. Onyenweaku1, John E. Mgbang1 and Ejiofor Charles E.2

1Department of Biochemistry, Faculty of Basic Medical Sciences, University of Calabar, P.M.B. 1115, Calabar, Nigeria

2Department of Veterinary, Parasitology, Faculty of Veterinary Medicine, University of Abuja, P.M.B. 117 Abuja, Nigeria

Pub. Date: April 13, 2017

Cite this paper:
Chinaka Nwaehujor, Eridiong O. Onyenweaku, John E. Mgbang and Ejiofor Charles E.. In vitro Free Radical Scavenging Activities of the Saponin-rich Fractions from Vernonia amygdalina Del. (Compositae). American Journal of Pharmacological Sciences. 2017; 5(1):11-17. doi: 10.12691/ajps-5-1-3

Abstract

Vernonia amygdalina leaves is consumed fresh or dried after removal of bitter taste by the peoples of the South-Eastern Nigeria, especially the Igbos. It is also widely used in cooking and serves as a vegetable and ethno-medicinal herb against many ailments. Not much is known of its in vitro antioxidant potentials, especially the major saponins and those of its fractions. In this study, the 80 % methanol extract of the leaves was fractionated by analytical TLC-guided column chromatography, preparative TLC and further purified with HPLC using gradient concentrations of solvent mixtures of chloroform, n-butanol, methanol and distilled water yielding VASC, VASB, VASM70, VASM65 and VASM50. Quantitative phytochemical analysis gave their % total saponin content as 55.8, 64.6, 21.4, 78.8 and 33.8 respectively. The 5 saponin-rich fractions (VASC, VASB, VASM70, VASM65 and VASM50) showed no significant signs of toxicity in mice at oral doses of 1500 mg/kg although VASM50 at 1500 mg/kg caused increased urination and watery fecal deposits which disappeared after 18hrs. The saponin-rich fractions of Vernonia amygdalina were evaluated for in vitro free radical scavenging properties using 2, 2-diphenyl-1-picrylhydrazyl (DPPH.), ferric reducing antioxidant power (FRAP), hydrogen peroxide scavenging assay, hydroxyl radical scavenging assay, ABTS radical cation scavenging activity assay, anti-lipid peroxidation assay, B-carotene bleaching assay and superoxide anion using ascorbic acid, butylated hydroxytoluene (BHT), catechin and gallic acid as reference standards. The results showed that the saponin-rich fractions from Vernonia amygdalina leaves had a potent DPPH radical scavenging and FRAP activities comparable to those of reference standards used. The IC50 values were also comparable to those of standards especially that of VASM65. Vernonia species are known to contain abundant saponins and flavonoids which are polar compounds and readily soluble in methanol. Most saponins and their derivatives of plant origin are known to possess great antioxidant potentials. This may explain the above observed antioxidant activities and thus, the use of the leaves in different traditional curative therapies in Eastern Nigeria, and as vegetable.

Keywords:
Vernonia amygdalina Saponins Antioxidants free radicals 2 2-diphenyl-1-picrylhydrazyl (DPPH) ferric reducing antioxidant power (FRAP)

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References:

[1]  Badarinath AV, Mallikarjuna RAK, Chethy CMC, Ramkanth S, Rajan TVS, Gnanaprakash K (2010). A review on in vitro antioxidant methods: Comparisms, correlations and considerations. Int J Pharm Tech Res, 2, 1276-1285.
 
[2]  Benzie IF and Strain, JJ (1999). Ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: The FRAP assay. Anal. Biochem. 239, 70-76.
 
[3]  Berhow MA, Wagner ED, Vaughn SF and Plewa MJ (2000). Characterisation and anti-mutagenic activityof soybean saponins. Mutat. Res. 448: 11-22.
 
[4]  Bokhari J, Khan MR, Shabbir M, Rashid U, Jan S, Zai JA (2013). Evaluation of diverse antioxidant activities of Galium aparine. Spectro Acta Part A: Mol. Biomol. Spectro. 102:24-29.
 
[5]  Cui MF, Song F, Zhou Y, Liu Z and Liu S (2000). Rapid identification of saponins in plant extracts by electro-spray ionisation multi-stage tandem mass spectrometry and liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom 14: 1280-1286.
 
[6]  Dorman HJD, Kosar M, Kahlos K, Holm Y, Hiltunen R (2003). Antioxidant properties and composition of aqueous extracts from Mentha species, hybrids, varieties, and cultivars. J. Agric. Food Chem. 51: 4563-4569.
 
[7]  Elzaawely AA, Xuan TD, Koyama H, Tawata S (2007) Antioxidant activity and contents of essential oil and phenolic compounds in flowers and seeds of A. zerumbet (Pers.) B.L. Burtt. & R.M. Sm. Food Chem. 104:1648-1653.
 
[8]  Focho DA, Nkeng EAP, Lucha CF, Ndam WT, Afegenui A (2009). Ethnobotanical survey of plantsused to treat diseases of the reproductive system and preliminary phytochemical screening of some species of Malvaceae in Ndop Central Sub-Division, Cameroon. Journal of Medicinal Plants Research 3, 301-314.
 
[9]  Francis G, Kerem Z, Harinder PSM and Becker K (2002). The biological action of saponins in animal systems: a review. Br. J. Nutr. 88:587-605.
 
[10]  Gestetner B, Assa Y, Henis Y, Birk Y and Bondi A (1971). Lucerne saponins. IV. Relationship between their chemical constitution, and haemolytic and antifungal activities. J. Sci. Food Agric. 22:168-172.
 
[11]  Gurfinkel DM and Rao AV (2002). Determination of saponins in legumes by direct densitometry. J. Agric. Food. Chem. 50:426-430.
 
[12]  Halliwell B, John G, Okezie I (1987). The deoxyribose method: a simple “test-tube” assay for determination of rate constants for reactions of hydroxyl radicals. Anal. Biochem. 165:215-219.
 
[13]  Harbourne JB (1998). Phytochemical methods: A guide to modern techniques of plant analysis. 2nd edition. Chapman and Hall, London, pp. 282.
 
[14]  Hostettmann K and Marston A (1995). Saponins. Cambridge University Press: Cambridge.
 
[15]  Harbourne JB (1973). Phytochemical Methods. Chapman and Hall, London, pp: 113.
 
[16]  Hostettmann M and Marston A (1986). High Pressure Liquid Chromatography: Preparative Chromatography Technigues. Springer-Verlag, New York Heidelberg, Berlin, pp: 56-61.
 
[17]  Igile GO, Oleszek W, Jurzysta MS, Burda Faunsho M, Fasanmade A (1994). Flavonoids from Vernonia amygdalina and their antioxidant activities. J. Agric Food Chem. 42(11), pp 2445-2448.
 
[18]  Kerem Z, German-Shashoua H, Yarden O (2005). Microwave-assisted extraction of bioactive saponins from chickpea (Cicer arietinum L) J Sci Food Agric 85:406-412.
 
[19]  Kuduo S, Tonomura M, Tsukamato C, Uchida T, Yoshikoshi M and Okubo K (1994). Structural elucidation and physiological properties of genuine soybean saponins. In Food Phytochemicals for Cancer Prevention I: Fruits and Vegetables.
 
[20]  Mandarino JMG, Carrao-Panizzi MC and Shiraiwa M (2000). Composition and content of saponins in soybean seeds of Brazilian cultivars and breeding lines. Proceedings of3rd InternationalSoybean Utilisation Conference, Tsukuba, Japan.
 
[21]  Mensor LL, Menezes FS, Leitao GG, Reis AS, Dos Santos TC, Coube CS, Leitao SG (2001). Screening of Brazilian plant extracts for antioxidant activity by the use of DPPH free radical method. Phytother. Res., 15, 127-130.
 
[22]  Nishikimi M, Rao NA, Yagi K (1972). The occurence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem. Biophys. Res. Commun. 46: 849-854.
 
[23]  Nwaehujor CO, Asuzu OV, Asuzu IU (2014) Membrane stability of red blood cells in diabetic mice treated with D-3-O-methylchiroinositol. Am. J. Pharmacol. Sci. 2(1): 24-26.
 
[24]  Nwaehujor CO, Ode JO and Akande MG (2012). In vitro antioxidant potentials of some herbal plantsfrom Southern Nigeria. J. Med. Sc., 13 (1): 56-61.
 
[25]  Oleszek W, Price KR, Colquhoun IJ, Jurzysta M, Ploszynski M and Fenwick GR (1990) Isolation and identification of alfalfa (Medicago sativa L) root saponins—their activity in relation to a fungal bioassay. J. Agric. Food Chem. 38:1810-1817.
 
[26]  Oleszek WA (2002). Chromatographic determination of plant saponins. J. Chromatogr. A 967:147-162
 
[27]  Price KR, Johnson IT and Fenwick GR (1987) The chemistry and biological significance of saponins in foods and feeding stuffs. CRC Crit. Rev. Food Sci. Nutr. 26: 27-135
 
[28]  Ramdas A, Seema M. (2010). Antioxidant activity and antimutagenic effect of phenolic compounds in Fermonia limonial (L) swingle fruit. International journal of pharmacy and pharmacology, 2, 68-73.
 
[29]  Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C: Antioxidant activity applying an improved ABTS radical cation decolorisation assay. Free Rad. Biol. Med. 1999, 26: 1231-1237.
 
[30]  Rosen RT (eds). American Chemical Society: Washington, DC; 340- 348.
 
[31]  Shiraiwa M, Harada K and Okubo K. 1991a. Composition and content of saponins in soybean seed according to variety, cultivation year, and maturity. Agric. Bio. Chem. 55: 323-331.
 
[32]  Shiraiwa M, Harada K and Okubo K. 1991b. Composition and structure of 'group B saponin' in soy seed. Agric. Bio. Chem. 55: 911-917.
 
[33]  Shiraiwa M, Kudo S, Shimoyamada M, Harada K and Okubo K. 1991. Composition and structure of 'group A saponin' in soybean seed. Agric. Bio. Chem. 55: 315-322.
 
[34]  Trease GE and Evans WC, 1984. Separation and isolation of plant constituents In: Trease and Evans Pharmacognosy 12th Ed. Baillere Tindall London, pp. 242-259.
 
[35]  Trouillas P, Calliste CA, Allias DP, Simon A, Marfak A, Delage C et al., 2003. Antioxidant, anti-inflammatory and antiproliferative properties of sixteen water plant extracts used in the Limousine countryside as herbal teas. Food Chem. 80: 399-407.
 
[36]  Umamaheswari M, Chatterjee T: In vitro antioxidant activities of the fractions of Coccinia grandis L. leaf extract. African J. Trad. Com. Alt. Med. 2008, 5: 61-73.