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Of Amaranthus Palmeri International Research Journal of Biological Sciences ________________ _______ ___________ E-ISSN 2278-3202 Vol. 5(5), 35-43, May (201 6) Int. Res. J. Biological Sci. Antioxidant Activities of Phytonutrient of Amaranthus Palmeri Rajeshwari A. 1* , Thammanna Gowda S.S. 2 and Somashekar G.N .3 1Department of Biochemistry, Adichunchanagiri Institute of Medical Sciences, B.G. Nagar, Ma ndya District, Karnataka. India 2Central Research Lab., Adichunchanagiri Institute of Medical Sciences., B.G. Nagar, Mandya District, Karnataka. India. B.G. N agar, Mandya District, Karnataka. India 3Department of Biochemistry, Adichunchanagiri Institute of Medical Sciences, B.G. Nagar, Mandya Di strict, Karnataka. India [email protected] Available online at: www.isca.in , www.isca.me Received 31 st March 2016, revised 04 th April 2016, accepted 10 th May 201 6 Abstract Amaranthus is commonly consumed green leafy vegetable in India. It is bushy perennial herb available throughout the year in India. Animal experiments and human studies have shown that, it is an effective hypoglycemic and hypocholesterolaemic agent. Amaranthus leaves rich in carbohydrates, proteins and vitamin A, vit K and vit C. It also contains minerals such as iron, magnesium, calcium and phosphorous. It reduces bad cholesterol and anemia. In the current study, we evaluated the water, methanol, and chloroform and h exane extract of Amaranthus in-terms of their antioxidant activity in -vitro models. Ours studies proved that, compared to all the other extracts, methanol and water extracts showed more activity. Among these two extracts, water extract possess highest anti oxidant activity in all the model systems. Lipid peroxidation activity of the was 75 and 66% in both linoleic and RBC membrane model system. Both the extracts were capable of scavenging peroxy and superoxide, hydroxylradicals and possessed strong reducing powers and ferrous iron chelating capabilities and their activity is comparable with that of standard antioxidants BHA and α-tocopherol. The proximate analysis revealed the presence of high concentration of phenolic compounds (180 -210µg) which may be the f act for maximum antioxidant activity. Keywords : Amaranthus, Hydroxyl Radicals, Superoxide, BHA, Methanol. Introduction and cerebrovascular disease and cancer mortality 8-10 . Sources of natural antioxidants are primarily plant phenolics that may occur Antioxidants eradicate harmful Reactive Oxygen Species in all parts of the plant 11 . Green leafy vegetables offer a cheap (ROS), free radicals and peroxides in our body by scavenging or but rich so urce of a number of micronutrients and other reducing their activity, otherwise they could damage DNA, cell phytochemicals having antioxidant properties. A lot of studies membranes and other cell components. Free radicals are the have been analyzed the antioxidant potential of a wide variety of natural by-products produced during many physiological vegetables including Cacao beans, potato, spinach and 1 activities in our body . Damages caused due to these ROS and legumen 12,13 . oth er free radicals to various tissues can be taken care by internal body antioxidants. When the damage is much severe, Plant derived natural products such as flavonoids, terpens and there must be need of external synthetic or natural antioxidants. alkaloids have received considerable attention in recent years There is much interest among food manufacturers in natural due to their pharmacological properties including cytotoxic antioxidants t o act as replacement for synthetic antioxidants hepatoprotective and cancer chemopreventive effects. The such as Butylated Hydroxy Anisole (BHA). Butylated growing inter est in the substitution of synthetic food Hydroxyl Toluene is commonly used by the industry to combat antioxidants by natural antioxidant and in health implications of formation of off-flavors and rancidity, particularly in fat based antioxidants in nutraceuticals has hastened the research on 2 products . There is also an em erging consensus that the vegetable sources and screening of raw materials for identifying micronutrients and non nutrient components such as various antioxidants. The antio xidants could attenuate this oxidative phytochemicals of fruits and vegetables play a preventive role in damage of the tissue in directly by enhancing natural defenses 3-6 the development of chronic diseases . of cell/or directly by scavenging the free radicals spices Dark green leafy vegetables are the sources of vitamins, Amaranth is a leaf vegetable and grain being consumed for miner als and carotenoids which act as antioxidants in the body. centuries all over the world. It be longs to the family Compounds present in dark green leafy vegetables can inhibit Amaranthaceae and the genus Amaranthus. All parts of the body 7 the growth of certain types of cancer . Epidemiological studies are edible which are highly nutritious. Amaranth seeds are used show that consumption of fruits and vegetables with high since ancient time in Central and Latin America amaranth phenolic conten t correlate with reduced blood pressure, cardio leaves are used across Asia. Amaranth grain is emergi ng into International Science Community Association 35 Research Journal of Biological Sciences _________________________________________________________ E- ISSN 2278-3202 Vol. 5(5), 35-43, May (2016) Int. Res. J. Biological Sci. the forefront among grains because of its remarkable nutritional procedure of Shimazaki et al and an assessment of oxidation values. It is rich in considerable amount of calcium, iron, was achieved by measurement of TBARS according to Dahle. et phosphorus and carotenoids, vitamins A, C and E. Oil is rich in al 16 . 6µmoles of linoleic acid (solubilized in Hexane) or 100µl unsaturated fatty acids, squalene, polyphenols, flavonoids and of erythrocyte ghost suspension (300µg membrane proteins) high in fiber three time that of wheat. were subjected to peroxidation by ferrous sulphate-ascorbate (10:100µmole) in 0.5 ml of Tris buffered saline TBS (10mM, The protein content of amaranth is 28.1 gms for a cup also rich pH 7.4, 0.15M NaCl). With or without extracts of Amaranthus in essential amino acids lysine and methionine. The leaves, (0.05 to 0.25 mg/0.5ml of TBS) 400µM BHA/α-tocopherol seeds and flowers of the plant are responsible for all of the serving as positive controls and contents were incubated for 1 health benefits 14 . Amaranth seeds and leaves have been used to hr, at 37 0C. The reaction was terminated by addition of 500µl treat hypercholesterolemia in animals studies as on astringent alcohol. The extent of lipid per oxidation was assayed in for stopping diarrhea bloody stools and urine. The medicinal triplicates by malondialdehyde formation using 1ml of 1% uses of the leaves flowers and seeds of amaranth are leaves are thiobarbituric acid. The contents were kept in boiling water used to neutralize acidity leaf poultice is used to treat snake and bath for 15 min, cooled then 2 ml of acetone is added to insect bites and to remove stings of bee wasp hornet scorpion. stabilize the color which is measured at 535 nm. Strong leaf decoction is used to treat diarrhea dysentery mouth irritations to remove worms and other parasites from GIT and Reducing power activity of Amaranthus: Reducing activity of throat irritation, wound. Amaranthus was tested according to the method of Oyaizu 17 . Various concentrations of the water and methanol extracts of Amaranthus palmeri is an edible flowering plant in the amaranth Amaranthus (0 to 500µg) were mixed with 200µl of 0.2M genus also known as palmeri amaranth palmeris pigweed and phosphate buffer, pH 6.5 and 200 µl of 1% potassium careless weed. The leaves, stems and seeds of palmeri amaranth ferricyanide, and then incubated at 50 0C for 20 min. 10% are edible and highly nutritious 15 . Palmeri amaranth is widely trichloroacetic acid (250µl) was added to the reaction mixture used for thousands of years in mexico north and south. However and centrifuged at 3000 g for 10 min at room temperature. The there is lack of scientific report regarding antioxidant properties resulting supernatant was taken and mixed with 500 µl of of Amaranthus palmeri, thus the present study provides the double distilled water and 100µl of 0.1% ferric chloride then scientific validation for the traditional uses and antioxidant incubated at 37 0C for 10 mins. The absorbance was measured at properties of the same. 700nm. Increased absorbance indicated increased reducing power. Materials and Methods 1, 1-Diphenyl-2- picryl hydrazyl (DPPH) radical Scavenging Materials: Amaranthus leaves were purchased from local farms Effect: DPPH radical scavenging activity was determined in Mandya, Karnataka. They were washed, dried at ambient according to the method of Schimada et al. Reactions were temperature for a week with frequent dispersion under sterile performed in 1ml of freshly prepared 1mM DPPH ethanolic conditions. The dried leaves were sieve powdered, sealed in solutions and various concentrations of water and methanol 0 plastic bags and stored at 4 C until use. extracts of Amaranthus (0-500µg). Reaction mixtures were incubated at 37 0C for 30 minutes and the absorbance at 517 nm Chemicals: All chemicals and reagents used were of analytical was measured. This assay was done in triplicates. grade, purchased from Merck. Pvt. Ltd., Bombay and Sigma- Aldrich. Co. (St. Louis, USA), unless otherwise indicates all of Hydroxyl Radicals scavenging activity: Deoxy ribose assay is the reagents were prepared in deionized water to eliminate the used to determine the hydroxyl radical scavenging activity contamination of metal ions. according to the method of Halliwell et al 19 . Reaction mixtures containing various concentrations of water and methanol Methods: Preparation of extracts: Dry Amaranthus (10 gm) extracts of Amaranthus,0.02M phosphate buffer, pH 7.4, 2mM was weighed and extracted with 100 ml of double distilled H2O2, 0.05mM Ferric chloride, 0.05mM ascorbate , 6mM water, methanol chloroform and hexane separately, vortexed for deoxy ribose and 0.05 mM EDTA were incubated at 37 oC for 5 hr at room temperature, then centrifuged at 10,000 rpm at 30 minutes.
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