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Antioxidant Activity, Total Phenolics and Flavonoid Contents of Some Edible Green Seaweeds from Northern Coasts of the Persian Gulf

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Antioxidant Activity, Total Phenolics and Flavonoid Contents of Some Edible Green Seaweeds from Northern Coasts of the Persian Gulf Iranian Journal of Pharmaceutical Research (2014), 13 (1): 163-170 Copyright © 2014 by School of Pharmacy Received: September 2012 Shaheed Beheshti University of Medical Sciences and Health Services Accepted: December 2013 Original Article Antioxidant Activity, Total Phenolics and Flavonoid Contents of some Edible Green Seaweeds from Northern Coasts of the Persian Gulf Massoumeh Farasata*, Ramazan-Ali Khavari-Nejada, Seyed Mohammad Bagher Nabavib and Foroogh Namjooyanc aDepartment of Biology , Science and Research Branch , Islamic Azad University,Tehran, Iran. bDepartment of Marine Biology, University of Marine Sciences and Technology, Khorramshahr, Iran. cMarine Pharmaceutical Research Center, Pharmacognosy Department, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Iran. Abstract The antioxidant activity, contents of total phenolics and flavonoids were quantified in the methanolic extracts of four Ulva species (Ulva clathrata (Roth) C.Agardh, Ulva linza Linnaeus, Ulva flexuosaWulfen and Ulva intestinalis Linnaeus) grown at different parts of northern coasts of the Persian Gulf in south of Iran. The seaweeds were collected from Dayyer, Taheri and Northern Ouli coasts in April 2011. Methanolic extracts of the seaweeds were assessed for their antioxidant activity using DPPH radical scavenging assay and was performed in a microplate reader. All species exhibited a DPPH radical scavenging activity, and among the species, -1 Ulva clathrata demonstrated greater antioxidant potential with a low IC50 (0.881 mg mL ) in comparison with those of the other species. Also the highest phenolic content (5.080 mg GAE g-1) and flavonoid content (33.094 mg RE g-1) were observed in U.clathrata. Total phenolic and flavonoid contents showed positive correlations with the DPPH radical scavenging activity (p < 0.01) and negative correlations with IC50 (p < 0.01).The results suggest that these edible green seaweeds possess antioxidant potential which could be considered for future applications in medicine, dietary supplements ,cosmetics or food industries. Keywords:Antioxidant activity; Total phenolics; Flavonoid; Seaweeds; Ulva. Introduction plants are rich sources of phytochemicals possessing important properties such as Free radicals have been claimed to antioxidant activity. Many investigators have play an important role in affecting human found several types of antioxidants from health by causing many diseases (e.g., heart different parts of various plant species such as diseases, cancer, hypertension, diabetes and oilseeds, cereal crops, vegetables and spices (2). atherosclerosis). In the past decade, antioxidants Recently, polyphenolic compounds including have shown their relevance in the prevention flavonoids is known as safe and non-toxic of various diseases, in which free radicals are antioxidants. Many studies have shown that a implicated (1). high dietary intake of natural phenolics is strongly According to the previous studies, terrestrial associated with longer life expectancy, reduced risk of developing some chronic diseases, various * Corresponding author: types of cancer, diabetes, obesity, improved E-mail: [email protected] endothelial function and reduced blood pressure Farasat M et al. / IJPR (2014), 13 (1): 163-170 (3-5). Phenolic compounds are commonly Once harvested, seaweeds were washed with found in plants and seaweeds. Like other plants, fresh water to remove sands, salts and epiphytes, seaweeds contain various inorganic and organic and then, were air-dried at room temperature with substances, which can benefit human health good controlled air condition carefully. The algae (6). It has been observed that ROS production samples were pressed and stored in 5% formol in algae is stimulated by various environmental for identification. Voucher specimens were stresses, such as high light levels, heavy metals, deposited in Jundishapur Marine Pharmaceutical high salt concentrations, UV radiation etc. Algae Research Center herbarium. Morphological generally has higher antioxidant activity due to and anatomical examinations of cell structures a higher contents of nonenzymatic antioxidant were done with the aid of stereomicroscope and components, such as ascorbic acid, reduced light microscope. The samples were identified glutathione, phenols and flavonoids (7). As according to the characteristics and identification a result, many marine bio-sources in the last keys in the taxonomic publications (11-15). decades have attracted attention in the search Samples kept at -50 ºC until experiments were for natural bioactive compounds to develop processed and milled into powder before new drugs and healthy foods. Compounds with extraction. antioxidant, antiviral, antifungal, antimicrobial, Dried seaweed sample powder (200 mg) antitumor and anti-inflammatory activities have was extracted with 6 mL 80% methanol in been found in brown, red and green algae (8). an ultrasonic bath for 20 min, vortexed for 30 The antioxidant activity of several seaweeds min and then left to stand at room temperature has been reported (9, 10). Ulva genus, an edible for 48 h. The extract centrifuged at 1500 g for seaweed, and an important food source in many 10 min, filtered through Watmann No.1 filter south-east Asian countries is also recognized by paper and then, was freeze dried. The dried its synonymous name as Enteromorpha. To the extracts were weighed and the yield of each best of our knowledge, there is no publication on extract was calculated. The stock solutions of the antioxidant activities of green seaweeds from the extracts were adjusted with 80% methanol Iran. The present study aimed to investigate the to final concentration of 2 mg (dry extract) mL-1. antioxidant properties of four Ulva species from Dilutions were made to obtain concentrations 1, the northern coasts of the Persian Gulf for future 0.5 and 0.1 mg mL-1. applications in medicine, dietary supplements, cosmetics or food industries. DPPH free radical scavenging activity DPPH radical scavenging activity was Experimental determined according to the method of Zhang et al. (2007) with slight modifications (16). Briefly, Chemicals 100 µL of each extract at various dilutions, were Ascorbic acid, Folin-ciocalteu reagent, Gallic mixed with 100 µL of 0.16 mM DPPH solution acid and Methanol were purchased from Merck .The mixture was vortexed for 1 min, kept for Company (Darmstadt, Germany). DPPH and 30 min in dark and then, the absorbance was Rutin were purchased from Sigma Chemical measured at 517 nm in an automated microplate Co (St.Louis, MO, USA). All the chemicals and reader (Sunrise-Elisa Reader, Tecan, Swiss). The reagents used were of analytical grade. antioxidant capacity was calculated using the following equation: Collection and preparing of algal extract The seaweeds were collected at low tide % Inhibition = (Acotrol - (Asample - Ablank) / Acotrol) × 100 time(according to the tide time table obtained from www.iranhydrography.org) along the Where the Acotrol is the absorbance of the northern coasts of the Persian Gulf, from Dayyer, control (DPPH without sample), the Asample is Taheri and Northern Ouli (Figure 1) in April the absorbance of the test sample (the sample 2011. The latitude and longitude of each sampling test and DPPH solution), and the Ablank is the location was recorded by GPS tracking device. absorbance of the sample blank (Sample 164 Antioxidant Activity, Total Phenolics and Flavonoid Contents Figure. 1. Study area. Figure. 1. Study area. without DPPHthe DPPH free radical solution). scavenging The half-maximalactivity reaction was recorded at 415 nm. The calibration inhibitory concentration (IC50) was calculated by curve was prepared by using Rutin methanolic - linear regressionDPPH radical analysis scavenging and expressed activity was as meandetermined solutions according at to concentrations the method of ofZhang 12.5 etto 100al. µg mL of three determinations. Ascorbic acid was used 1. FC was expressed as mg Rutin equivalents per as positive(2007) control. with slight modifications (16). Briefly, 100 gramµL of ofeach dried extract extract at various (mg dilutiRE gons,-1). were Determinationmixed with 100of totalµL of phenolic0.16 mM DPPHcompounds solution .The mixturStatisticse was vortexed for 1 min, kept for 30 and flavonoidmin in dark content and then, the absorbance was measured at 517Data nm werein an automatedexpressed microplate as means reader ± standard Total phenolic compounds (TPC) of algal errors of three replicate determinations. All extracts (Sunrise-was determinedElisa Reader, byTecan Folin-Ciocalteu, Swiss). The antioxidantstatistics capacity analyses was were calculated carried using out usingthe SPSS reagent according to the method of Antolovich 16.0 for Windows. To determine whether there et al. (2002)following (17) equation: with minor modifications. In were any differences among the means, one Brief, 20 µL of extracts were mixed with 100 way analysis (ANOVA) and the Duncan’s new µL of 1:10% Inhibition Folin-Ciocalteu = (Acotrol - reagent(Asample - followedAblank) / A cotrolby ) × 100multiple range test were applied to the result. the addition of Na2CO3 (80 µL, 7.5%). The assay p-values < 0.05 were regarded to be significant. was carriedWhere out the in A microplate.cotrol is the absorbance After incubation of the control The (DPPH Pearson without correlation sample), theanalysis Asample was is the performed at room temperature for 2 hours in dark, the between antioxidant activity and total phenolic absorbanceabsorbance at 600
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