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Antioxidative Activity of Sixty Plants from Iran

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Antioxidative Activity of Sixty Plants from Iran Iranian Journal of Pharmaceutical Research (2004) 3: 55-59 Received: December 2003 Accepted: February 2004 Original Article Antioxidative Activity of Sixty Plants from Iran Effat Souria, Gholamreza Aminb, Anahita Dehmobed-Sharifabadic, Atefeh Nazifia, Hassan Farsama* aDepartment of Medicinal chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran. bDepartment of Pharmacognosy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran. cDepartment of Pharmacognosy, School of Pharmacy, Shaheed Beheshti University of Medical Sciences, Tehran, Iran Abstract Antioxidants are vital substances which possess the ability to protect the body from damage caused by free radical-induced oxidative stress. A variety of free radical scavenging antioxidants exist within the body which many of them are derived from dietary sources like fruits, vegetables and teas. This article describes a test method for screening the antioxidant activity of 60 Iranian plants of Iran by linoleic acid peroxidation test using 1, 3-diethyl-2- thiobarbituric acid as the reagent. Some plants including Achillea wilhelmsii, Berberis crataegina, Buxus hyrcana, Chrysanthemum cinerariaefolium, Colutea persica, Hyoscyamus niger, Mentha pulegium, Nerium oleander, Pteropyrum aucheri, Rhus coriaria, Rosa canina, Scutellaria pinnatifida, Thymus pubescens, Verbascum alceoides and Ziziphora clinopodioides subsp. rigida showed antioxidant activity (0.41<IC50<1.64 µg) comparable to α-tocopherol (IC50=0.60 µg), which was used as the positive control. Keywords: Plant; Antioxidant; Linoleic acid; 1, 3-Diethyl-2-thiobarbituric acid. Introduction on the use of synthetic antioxidants due to their probable side-effects has increased the In recent years, it has been established that contribution of natural antioxidants (8). free radicals and oxidative stress are involved in The antioxidant activity of several plant the pathophysiology of a variety of disorders constituents, beyond the vitamins, in the form including atherosclerosis, chronic renal failure, of crude extract or isolated compound has been diabetes mellitus, cancer, immune dysfunction put widely into consideration (8-10). and aging (1-6). In relation to these findings an Antioxidant activity of many phenolic extensive range of antioxidants both exogenous compounds, including flavonoids, has attracted and endogenous, whether synthetic or natural considerable attention and reported to be more have been presented for the treatment or powerful antioxidants than vitamins C, E and β- prophylaxis of disorders attributed to free carotene which are largely in routine use (11). radical oxidative damages (3, 4, 7). Restriction Consumption of the flavonoids and their * Corresponding author: potential significance as antagonists of E-mail: [email protected] oxidative stress has been the interesting subject E Souri, G Amin, A Dehmobed-Sharifabadi, A Nazifi, H Farsam / IJPR 2004, 3: 55-59 of many investigations (8, 9, 11, 12). Vegetables and evaporated to dryness under reduced and fruits are also reported to decrease the risk pressure in a rotary evaporator. The extracts of degenerative diseases and could have a then transferred to vials, kept at 4°C and protective effect against oxidative stress (11). examined for antioxidant activity. Antioxidants are also important for food protection against deterioration reactions caused Measurment of antioxidant activity by atmospheric oxygen (8). Considerable effort The potential of plant extracts to inhibit has been directed in search for safe antioxidants peroxidation of linoleic acid was assessed based from natural sources. Naturally occurring on a procedure described by Furuta et al. (13). antioxidants could be found in fruits, α-Tocopherol was used as the reference vegetables, nuts, seeds, leaves, flowers, roots compound. For a typical assay three dilutions of and barks. Many investigators have found each extract (0.02, 0.2 and 2 mg/mL) were different types of antioxidants in various kinds prepared. An aliquot of 20 µL of each dilution of plants (8-12). (equal to 0.4, 4 and 40 µg of extract) was mixed One of the best approaches for discovering with 20 µL of linoleic acid (2 mg/mL in new antioxidants is the screening of plant ethanol) and incubated at 80°C for 60 min. extracts. This study was carried out as part of a Incubated samples were cooled in an ice bath, project to investigate the antioxidant activity of followed by the addition of 200 µL of 20 mM 60 selected plants growing in Iran, against BHT, 200 µL of 8% SDS and 400 µL of linoleic acid peroxidation. distilled water. After mixing, 3.2 mL of 12.5 mM DETBA in sodium phosphate buffer (0.125 Materials and Methods M, pH 3.0) (warmed to 50°C) was added, mixed and heated at 95°C for 15 min. The Plant material mixture was cooled in an ice bath, 4 mL of The plants were collected from different ethyl acetate was added to each tube, vortexed regions of Iran. Information regarding the to extract the pink adduct from the aqueous collection of plants is mentioned in table 1. phase and centrifuged at 1500 rpm for 10 min Voucher specimens of all plants were deposited (F ). A control containing linoleic acid and at the Herbarium of the Faculty of Pharmacy, 1 other additives without antioxidants, Tehran University of Medical Sciences. The representing 100% lipid peroxidation, was also aerial parts were separated, air dried in the prepared (F ). The blank F and F solutions shade, powdered and kept in tightened light- 2 1 2 were prepared as described above but without protected containers. linoleic acid. The fluorescence intensity of F 1 and F samples was measured against their Chemicals 2 blanks (F and F respectively) at an excitation Linoleic acid, 1, 3-diethyl-2-thiobarbituric 3 4 wavelength of 515 nm and an emission acid (DETBA) and quercetin dihydrate were wavelength of 555 nm in a spectrofluorimeter obtained from Merck (Darmstadt, Germany), (Model RF-5000, Schimadzu, Kyoto, Japan). Aldrich Chemical Co. (Milwaukee, WI, USA) The antioxidant activity was calculated as the and Fluka Chemical Co. (Buchs, Switzerland) percentage of peroxidation inhibition using the respectively. α-Tocopherol, sodium dodecyl following equation (14): sulfate (SDS) and butylated hydroxytoluene (BHT) were purchased from Sigma Chemical % of peroxidation inhibition= [1-(F1-F3)/(F2-F4)]×100 Co. (St. Louis, MO, USA). All other chemicals (equation 1) and solvents were of analytical grade from Merck. All extracts and the reference substance were assayed at least at three concentrations in Extraction triplicates and the results were averaged. A A quantity (50 g) of each powdered plant percentage inhibition vs log concentration curve was extracted in a Soxhlet apparatus with 80% was drawn and the concentration of sample methanol. The methanolic extracts were filtered which is required for 50 % inhibition was 56 Antioxidative activity of sixty plants from iran determined by linear interpolation and of 60) showed more than 80% peroxidation expressed as the IC50 value. inhibition, using 40 µg of plant extract in the reaction mixture. Plant extracts including Results and Discussion Achillea wilhelmsii, Buxux hyrcana, Chrysanthemum cinerariaefolium, Colutea The botanical characteristics of studied persica, Hyoscyamus niger, Mentha pulegium, plants and inhibitory effect of their methanolic Myrtus communis, Nerium oleander, Paliurus extracts on linoleic acid peroxidation are spina-christi, Peganum harmala, Pterocarya provided in table 1 in alphabetical order. As fraxinifolia, Rhus coriaria, Rosa canina, Smilax displayed in table 1, most plant extracts (44 out excelsa, Thymus migricus, Thymus pubescens, Table 1. The botanical names and antioxidant activities of aerial parts of 60 plant extracts from Iran Peroxidation inhibition(%) Voucher No. Location Date* Alt* Scientific name Family name 0.4 µg 4 µg 40 µg IC (µg) No. 50 1 N. of Karadj 99/6/6 1300 6570-TEH Achillea wilhelmsii C. Koch Compositae 27.37±2.66 78.93±0.65 75.27±4.05 1.41± 0.27 2 E. of Karadj 99/6/9 1200 6586-TEH Achillea tenuifolia Lam. Compositae 28.09±4.19 60.57±3.20 88.06±3.14 1.88± 0.37 3 E. of Karadj 99/6/9 1120 6585-TEH Acroptilon repens (L.) DC. subsp. repens Compositae 26.72±5.06 46.85±0.39 85.88±1.82 4.68± 0.13 4 Vaysar, S. of Chalus 99/6/4 600 6567-TEH Asperula stylosa Trin Rubiaceae 15.56±4.20 48.38±2.15 82.65±3.294.51± 0.57 5 E. of Tehran 99/7/21 1100 6587-TEH Astrodaucus orientalis (L.) Drude Umbelliferae 22.28±2.65 58.09±1.63 83.11±2.822.86± 0.27 6 Margun, W. of Shiraz 99/7/22 2000 6619-TEH Ballota aucheri Boiss Labiatae 2.65±1.63 47.92±1.86 90.54±4.575.09± 0.88 7 Ruin, Khorassan 99/6/12 1500 6590-TEH Berberis crataegina DC. Berberidaceae 49.95±4.65 52.02±3.07 90.29±2.180.81± 0.30 8 Talesh N. of Iran 99/6/8 930 6582-TEH Buxus hyrcana Pojark Buxaceae 35.23±6.83 71.92±8.31 86.94±2.791.15± 0.45 9 W. of Karadj 99/6/4 1100 6561-TEH Capparis spinosa L. Capparidaceae 20.61±0.69 49.29±6.78 64.58±6.835.84± 2.85 10 E. of Tehran 99/6/21 1100 6596-TEH Carthamus oxyacantha M.B. Compositae 27.05±0.15 28.35±2.18 71.27±2.509.11± 0.92 11 Ruin, Khorassan 99/6/12 1500 6616-TEH Centaurea bruguieriana (DC.) Hand.-Mzt. subsp. Compositae 25.63±7.44 62.98±0.37 91.00±5.65 1.74±0.24 belangerana (DC.) Bornm. 12 E. of Tehran 99/6/21 1100 6598-TEH Chenopodium botrys L. Chenopodiaceae 31.72±1.05 36.26±4.07 51.59±2.57 45.76±14.4 13 Km.40 Lowshan – Rasht 99/6/8 1350 6583-TEH Chrysanthemum cinerariaefolium ( Trev ) Vis.
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