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Phytochemical Screening, Phenolic Content and Antioxidant Activity of Wild Date Palm (Phoenix Sylvestris Roxb.) Fruit Extracted with Different Solvents

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Phytochemical Screening, Phenolic Content and Antioxidant Activity of Wild Date Palm (Phoenix Sylvestris Roxb.) Fruit Extracted with Different Solvents International Food Research Journal 24(6): 2534-2542 (December 2017) Journal homepage: http://www.ifrj.upm.edu.my Phytochemical screening, phenolic content and antioxidant activity of wild date palm (Phoenix sylvestris Roxb.) fruit extracted with different solvents Saha, S., Barua, B. and *Sikdar, D. Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong – 4331, Bangladesh Article history Abstract Received: 30 August 2016 Fruits are known to have different levels of antioxidant capacity due to the presence of multiple Received in revised form: types of phenolic compounds in varying amounts. In this study phytochemical constituents, 10 October 2016 phenolic content, flavonoid content, and antioxidant activity of Phoenix sylvestris fruit were Accepted: 11 October 2016 investigated. Three solvents (methanol, ethanol, and acetone) with different concentrations (50%, 70% and 100%) and water were used for extraction. Phytochemical analyses showed the presence of tanins, flavonoids and saponins. Fruits extracted with 70% acetone showed highest Keywords total phenolic content (1554.85 mg GAE/100 g FW) and total flavonoid content (2121.77 mg CEQ/100 g FW) whereas lowest total phenolic content (212.94 mg GAE/100 g FW) and lowest Antioxidant activities total flavonoid content (130.41 mg CEQ/100 g FW) were found in 2H O extract. Antioxidant Phenolic content activities of different solvent extracts were evaluated using ferric reducing antioxidant power Phoenix sylvestris (FRAP) assay, total antioxidant capacity assay, reducing power assay and 2,2-diphenyl-1- Phytochemical screening picrylhydrazyl (DPPH) free radical-scavenging assays. With all the assay methods, 70% Solvent effect acetone extract showed the highest antioxidant capacity. It was also observed that the phenolic content were strongly correlated with the antioxidant activity. © All Rights Reserved Introduction inflammation, allergy, viral infection, and cancer (Middleton, 1998). Human body is characterized by continuous The phenolic composition, content and production of free radicals and other reactive oxygen distribution of fruits varies with ripeness, cultivar species due to aerobic metabolism. At the same specificities, cultural practices, geographic origin, time antioxidants and antioxidant enzymes exert growing season and postharvest storage conditions synergistic action in removing the free radicals (Gul et al., 1998; Miletic et al., 2012). Different (Uttara et al., 2009). Under physiological conditions methods are used for the extraction of phenolic oxidising agents and antioxidant defenses are in compounds from different parts of plant. These balance. However, if the production of free radicals methods differ in solvent system and condition exceeds the antioxidant capacity of a living system, used. Solvents such as water and aqueous mixtures it leads to several physiological and pathological of methanol, ethanol, acetone and ethyl acetate have abnormalities such as inflammation, gastric ulcer, been used for the extraction of polyphenols from cardiovascular disease, cancer, neuro-degenerative plant materials (Wizekoon et al., 2011; Chavan and ailments, diabetes mellitus, AIDS and aging (Jacob Amarowicz, 2013). The extraction method also and Burri, 1996; Stangeland et al., 2009; Ismail et plays a crucial role in estimating accurate antioxidant al., 2010). content and capacity. So it is very difficult to compare According to the polyphenol content, different data from the literature reports (Asami et al., 2003; fruits show different antioxidant capacities (Saura- Alothman et al., 2009; Do et al., 2014). calixto and Goni, 2006) and recent research has Phoenix sylvestris Roxb., together with 13 other confirmed that vegetables and fruits play an species, forms the genus Phoenix. In a phylogenetic important role in the prevention and treatment of study combining morphological, anatomical and different diseases caused by oxidative damage (He genetic data, it appears close to the date palm et al., 2006; Galeone et al., 2007; Hung et al., 2015). (Phoenix dactylifera L.) and to Phoenix theophrasti. Flavonoids are the largest group of phenols found in P. sylvestris is widely distributed in South Asia, fruits, vegetables, grains, bark, roots, stems, flowers, from Pakistan to Myanmar, across India, Nepal, tea, and wine and play a role against oxidative stress, Bhutan and Bangladesh (Barrow, 1995). This palm *Corresponding author. Email: [email protected] 2535 Saha et al./IFRJ 24(6): 2534-2542 produces edible fruits but it is generally called, “wild chloride and sodium carbonate were purchased from date palm” to distinguish it from the closely related R and M Chemicals (Essex, UK). Ascorbic acid, P. dactylifera which is known as “date palm” and is potassium ferricyanide [K3Fe(CN)6], 2,4,6-Tris cultivated agriculturally as the commercial source of (1-pyridyl)-5-triazine (TPTZ), DPPH, Gallic acid dates (Bharti, 2015). The fruit of P. sylvestris Roxb., and catechin were supplied by Sigma–Aldrich (St. locally known as Khejur and ripened in the month of Louis, MO, USA). All chemicals and reagents used June-July, is underutilized in Bangladesh. It is cooling, in the study were of analytical grade. oleaginous, cadiotonic, fattening, constipative, good in heart complaints, abdominal complaints, fevers, Extraction of phenolic compounds with different vomiting and loss of consciousness (Parmar and solvents Kaushal, 1982). The fruit is also used to treat back First, the fruit was cleaned with distilled water pain, stomachache, toothache, headache, arthritis, and wiped with paper towel and weighed whole after pain of buttocks, piles, nervous debility, and as removing the seeds. The edible portions were cut into nervine tonic, restorative, sedative in ethnomedicine. small pieces and stored at -40°C before lyophilization Methanol extract of this fruit possesses strong central using a freeze dryer. After lyophilization the fruits and peripheral antinociceptive activity as well as CNS were pulverized into powder form with a grinder and depressant, sedative and anxiolytic activity (Shajib et kept at -40°C before analysis. Then 2 g fruit samples al., 2015). There are few studies on its antioxidant were mixed separately with 20 ml of three different and pharmacological properties. Previously our solvents-water extraction systems (methanol, ethanol research group (Aklima et al., 2014) showed highest and acetone) at three different concentrations (50, phenolic content and antioxidative activities of 70 and 100%), and distilled H2O. Each mixture was this fruit among five Bangladeshi fruits by DPPH stirred with magnetic stirrer at room temperature and ferric reducing methods using 70% ethanol as (250C) for 4 hours and filtered with Whatman No. solvent whereas Prakash et al. (2013) determined 41 filter paper. The residues remained after filtration the phenolic content and antioxidative activities of P. were again treated in the same way and the resulting sylvestris fruit and leaf by same methods using 50% two filtrates for each solvent were mixed and stored methanol. However effects of solvent systems on the at 4°C. All the analyses were done within three days extraction of phenols and antioxidant activities of the of extraction. fruit have not been reported. The main objectives of this study were to Phytochemical screening of the fruit extract determine the phenolic contents and detail antioxidant Different solvent extracts were used for activities of the P. sylvestris fruit grown in Bangladesh phytochemical screening of compounds which and to examine the efficiency of different solvent include tannins, flavonoids, alkaloids and saponins systems for the extraction of phenolic compounds. according to the methods of Harborne (1998) and The phenolic constituents were extracted using three Trease and Evans (1989) with slight modifications. solvents, namely, methanol, ethanol and acetone with different proportions of water. Test for tannins Exactly 1.0 ml of fruit extract for each solvent Materials and Methods was dissolved in 10 ml of distilled water and filtered (using Whatman No. 1 filter paper). A blue coloration Plant materials resulting from the addition of ferric chloride reagent Fresh fruit sample without any physical and to the filtrate indicated the presence of tannins in the microbial damage were collected from Chittagong extract. region (Bangladesh) at maturation stage and the species was identified consulting Siddiqui et al. Test for alkaloids (2007). The sample was stored at -20°C prior to 0.2 ml of extract for each solvent was taken in a analysis. conical flask and ammonia solution (3 ml) was added. It was allowed to stand for few minutes to evaluate Chemicals and reagents free alkaloids. Chloroform (10 ml) was added to the Folin–Ciocalteu’s (FC) phenol reagent, sulphuric conical flask, shaken by hand, and then filtered. The acid, hydrochloric acid, methanol, acetone and chloroform was evaporated from the crude extract by ethanol were from Merck (Darmstadt, Germany). water bath and Mayer’s reagent (3 ml) was added. A Sodium nitrite, sodium acetate, aluminium chloride, cream color precipitation was obtained immediately ammonium phosphate, ammonium molybdate, ferric that showed the presence of alkaloids. Saha et al./IFRJ 24(6): 2534-2542 2536 Test for flavonoids solution at pH 5.5, 1 ml of 0.2 mM DPPH in ethanol, 1.0 ml of each solvent extract was taken in a and 1.492 ml of a 50% (v/v) ethanol aqueous solution. test tube and few drop of dilute NaOH solution
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