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Assessing the Polyphenolic, Nutritive and Biological Activities of Acetone, Methanol and Aqueous Extracts of Rumex Sagittatus Thunb

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Assessing the Polyphenolic, Nutritive and Biological Activities of Acetone, Methanol and Aqueous Extracts of Rumex Sagittatus Thunb African Journal of Pharmacy and Pharmacology Vol. 4(9), pp. 629-635, September 2010 Available online http://www.academicjournals.org/ajpp ISSN 1996-0816 ©2010 Academic Journals Full Length Research Paper Assessing the polyphenolic, nutritive and biological activities of acetone, methanol and aqueous extracts of Rumex sagittatus Thunb. F. O. Jimoh1, A. A. Adedapo2* and A. J. Afolayan1 1Department of Botany, University of Fort Hare, Alice 5700, South Africa. 2Department of Veterinary Physiology, Biochemistry and Pharmacology, University of Ibadan, Ibadan, Nigeria. Accepted 23 July, 2010 The nutritional, phytochemical and antioxidant activities of acetone, methanol and aqueous extracts of the leaves of Rumex sagittatus Thunb. were investigated using standard analytical methods in order to assess the therapeutical potential of the leaves of this plant. The proximate analysis showed that the leaves contained appreciable amount of ash, crude protein, lipids, fibre and carbohydrates. Elemental analysis in mg/100 g (DW) indicated that the leaves contained appreciable sodium, potassium, calcium, magnesium, iron, zinc, phosphorus, copper, manganese and nitrogen. The chemical composition in mg/100 g (DW) showed the presence of alkaloid, saponins and phytate. The extracts also exhibited DPPH and ABTS radical scavenging activities, which were comparable to that of ascorbic acid and BHT. Extracts also contained appreciable levels of polyphenolic compounds. In comparing the nutrient and chemical constituents with recommended dietary allowance (RDA) values, the results reveal that the leaves contain an appreciable amount of nutrients, minerals, and phytochemicals and low levels of toxicants. Key words: Antioxidant activities, Rumex sagittatus, nutritional value. INTRODUCTION Rumex sagittatus Thunb. (Polygonaceae) belong to a qualities. In Western Europe, dock leaves are a genus of about 200 species. The plant is native to traditional remedy for the sting of nettles, and suitable southern Africa, where it occurs from Malawi and Zambia larger docks such as Rumex obtusifolia often grown (south) to South Africa. It has become naturalized in conveniently in similar habitats as compared to Urtica many parts of Australia near urban areas, from dioica. It has been shown that dock leaf has high levels of Queensland to eastern Tasmania and warmer locales in chlorphenamine, a powerful antihistamine, finally, New Zealand (Arnold and De Wets, 1993; Wolf, 1999). providing hard evidence for the medicative power of R. The docks and sorrels, genus Rumex L., are a genus of patientia (Bicker t al., 2009). about 200 species of annual, biennial and perennial Many workers have reported the compositional herbs in the buckwheat family Polygonaceae. Members evaluation and functional properties of various types of of this family are very common perennial herbs growing edible wild plants in use in the developing countries in acidic, sour soils mainly in the northern hemisphere, (Lockeett et al., 2000; Akindahunsi and Salawu, 2005; but have been introduced almost everywhere. Many are Edeoga et al., 2006). The consumption of vegetables nuisance weeds (and are sometimes called dockweed or particularly plants containing functional phytochemicals dock weed), but some (notably the Common Sorrel, has also been linked to reduction in the incidence of Rumex acetosa) have edible leaves, used in soups and oxidative-stress-related diseases (Akubugwo et al., salads. The leaves of most species contain oxalic acid 2007). and tannin and many have astringent and slightly purgetive New developments in biopharmaceuticals point to the involvement of reactive oxygen species (ROS) in many diseases (Behera et al., 2006; Adedapo et al., 2008). Free radicals attack the unsaturated fatty acids in the *Corresponding author. E-mail: [email protected]. biomembranes resulting in membrane fluidity (Dean and 630 Afr. J. Pharm. Pharmacol. David, 1993). Free radicals have been implicated in the diluted with water 1:10 v/v) and 4 ml (75 g/l) of sodium carbonate. causation of several degenerative diseases and The tubes were vortexed for 15 s and allowed to stand for 30 min at compounds that can scavenge free radicals have great 40˚C for color development. Absorbance was then measured at 765 nm using the Hewlett Packard UV-VS spectrophotometer. Samples potential in ameliorating these disease processes (Di of extract were evaluated at a final concentration of 0.1 mg/ml. Matteo and Esposito, 2003; Behera et al., 2006). Plants Total phenolic content were expressed as mg/g tannic acid containing flavonoids have been reported to possess equivalent using the following equation based on the calibration 2 strong antioxidant properties (Raj and Shalini, 1999; curve: y = 0.1216x, R = 0.9365, where x is the absorbance and y is Badami et al., 2003; Behera et al., 2006; Tripathi et al., the tannic acid equivalent (mg/g). 2007). It is surprising that few indigenous vegetables species Determination of total flavonoids were consumed in South Africa by the general populace. The interview conducted during the course of our Total flavonoids were estimated using the method of Ordoñez et al. research in Alice and its surrounding villages indicated (2006). To 0.5 ml of sample, 0.5 ml of 2% AlCl3 ethanol solution that many of these species, though known, are was added. After one hour at room temperature, the absorbance was measured at 420 nm. A yellow color indicated the presence of considered as weeds and were not eaten by the people. flavonoids. Extract samples were evaluated at a final concentration This is in spite of the fact that these vegetables grow of 0.1 mg/ml. Total flavonoid content was calculated as quercetin spontaneously and in abundance around the rural (mg/g) equivalents using the following equation based on the homesteads. R. sagittatus was one of the wild vegetables calibration curve: y = 0.0255x, R2 = 0.9812, where x is the identified and the study was therefore, aimed at absorbance and y is the quercetin equivalent (mg/g). assessing its nutritional quality as well as antioxidant activities of the some extracts of the plant. Determination of total flavonols Total flavonols in the plant extracts were estimated using the MATERIALS AND METHODS method of Kumaran and Karunakaran (2006). To 2.0 ml of sample (standard), 2.0 ml of 2% AlCl3 ethanol and 3.0 ml (50 g/L) sodium Plant collection and extract preparation acetate solutions were added. The absorption at 440 nm was read after 2.5 h at 20˚C. Extract samples were evaluated at a final Fresh leaves of R. sagittatus were collected in November 2006 concentration of 0.1 mg/ml. Total flavonoid content was calculated around the University of Fort Hare campus Alice, South Africa as quercetin (mg/g) equivalents using the following equation based (30°00-34° 15´S; 22° 45´ -30° 15´E). These areas consist of on the calibration curve: y = 0.0255x, R2 = 0.9812, where x is the villages, which are generally classified as rural and poor. Professor absorbance and y is the quercetin equivalent (mg/g). D. Grierson of the Department of Botany, University of Fort Hare, authenticated the species. A voucher specimen was prepared and deposited in the herbarium of the Department of Botany (Jimoh The quantitation of total proanthocyanidins Med. 2006/710B). The plant material was allowed to air-dry at ambient temperature (±24˚C) and then milled. Twenty grams per The quantitation of proanthocyanidin was based on the procedure sample was extracted from 200 ml of acetone, methanol, and reported by Sun et al. (1998). A volume of 0.5 ml of 0.1 mg/ml of water, respectively, with stirring at ambient temperature for 18 to 24 extract solution was mixed with 3 ml of 4% vanillin-methanol h. Each extract was filtered using Whatman no. 1 filter paper and solution and 1.5 ml hydrochloric acid; the mixture was allowed to concentrated under reduced pressure to dryness below 40˚C. The stand for 15 min. The absorbance was measured at 500 nm. aqueous extract was freeze-dried. The extract yields (w/w) were Extract samples were evaluated at a final concentration of 0.1 acetone (3.2%), methanol (8.9%), and aqueous (9.3%). The dried mg/ml. Total proanthocyanidin content were expressed as catechin extracts thus obtained were used directly for the determination of equivalents (mg/g) using the following equation based on the the antioxidant activities (Taylor et al., 1996). calibration curve: y = 0.5825x, R2 = 0.9277, where x is the absorbance and y is the catechin equivalent (mg/g). Chemicals Determination of antioxidant activity 1,1-Diphenyl-2-picrylhydrazyl (DPPH), 2,2-azinobis-3- ethylbenzothiazoline-6-sulfonic acid (ABTS), 3-(2-pyridyl)-5,6- DPPH radical scavenging assay diphenyl-1,2,4-triazine-4,4-disulfonic acid, potassium ferricyanide; catechin, butylated hydroxytoluene (BHT), ascorbic acid, catechin, The effect of extracts on DPPH radical was estimated using the tannic acid, quercetin, TPTZ and FeCl3 were purchased from method of Liyana-Pathirana and Shahidi (2003). A solution of 0.135 Sigma Chemical Co. (St. Louis, MO, USA)., vanillin from BDH; mM DPPH in methanol was prepared and 1.0 ml of this solution Folin-Ciocalteus’s phenol reagent and sodium carbonate were from was mixed with 1.0 ml of extract in methanol containing 0.02 to 0.1 Merck Chemical Supplies (Darmstadt, Germany). All the other mg of the extract. The reaction mixture was vortexed thoroughly chemicals used including the solvents, were of analytical grade. and left in the dark at room temperature for 30 min. The absorbance of the mixture was measured spectrophotometrically at 517 nm. Ascorbic acid and BHT were used as references. The Determination of total phenolics ability to scavenge DPPH radical was calculated by the following equation: DPPH radical scavenging activity (%) = [(Abscontrol - Total phenol contents in the extracts were determined by the Abssample)]/(Abscontrol)] × 100 where Abscontrol is the absorbance of modified Folin-Ciocalteu method (Wolfe et al., 2003).
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