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Evaluation of Total Polyphenols, Flavonoids and Antioxidant Activity of Myrica Esculenta Buch-Ham.Ex D.Don: Fruits

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Evaluation of Total Polyphenols, Flavonoids and Antioxidant Activity of Myrica Esculenta Buch-Ham.Ex D.Don: Fruits wjpmr, 2021,7(2), 186-192 SJIF Impact Factor: 5.922 WORLD JOURNAL OF PHARMACEUTICAL Research Article Nishat et al. World Journal of Pharmaceutical and Medical Research AND MEDICAL RESEARCH ISSN 2455-3301 www.wjpmr.com Wjpmr EVALUATION OF TOTAL POLYPHENOLS, FLAVONOIDS AND ANTIOXIDANT ACTIVITY OF MYRICA ESCULENTA BUCH-HAM.EX D.DON: FRUITS Dr. Nishat Anjum1* and Y. C. Tripathi2 1Science Department, Ras Bihari Bose, Subharti University, Dehradun-248007. 2Chemistry and Bioprospecting Division, Forest Research Institute, PO New Forest, Dehradun – 248006, Uttarakhand, India. *Corresponding Author: Dr. Nishat Anjum Science Department, Ras Bihari Bose, Subharti University, Dehradun-248007. Article Received on 16/12/2020 Article Revised on 06/01/2021 Article Accepted on 26/01/2021 ABSTRACT Background: Fruits of Myrica esculenta belongs to Family Myricaceae, Fruits have been used to treat a variety of diseases for thousands of years. It is widely used in folk medicine to treat; a lot of research has been finding natural antioxidants of fruits of Myrica esculenta. The present study was aimed to evaluate the total polyphenol and antioxidant activity of Myrica esculenta. Methods: The present study was standard methods of phytochemicals. Total phenolic content the various extracts was determined spectrometrically using a modified Folin-Ciocalteu method and antioxidant efficacy by following DPPH radical scavenging protocol, total flavonoid content by reported method. Results: The results showed that Myrica esculenta of Phytochemical screening the presence of alkaloids, steroids, terpenoids, flavonoids, phenolics, tannins, saponins, glycosides, carbohydrate, protein and amino acids in different extracts of Myrica esculenta fruits. Fruits were recorded methanol extract maximum number of chemical constituents. High polarity solvent was present of flavonoids and phenolic constituents mostly found in extract. The total phenolic content (7.12±0.42 mg GAE/g of extract), total flavonoid content (4.54±0.017 mg QE/g of extract) and antioxidant activity (IC50, 55.00±0.341 µg/ml) were found highest in methanol extract. Conclusions: The present study established fruits of Myrica esculenta as rich sources of phenolic compounds and natural antioxidants. KEYWORDS: Myrica esculenta, Fruits, Phytochemicals, Phenolics, Flavonoids, Antioxidant activity. 1. INTRODUCTION is an evergreen dioecous tree, it is shade giving, medium size tree and it traditionally eats whole-seed and all Oxygen through excessive production of reactive oxygen fruits. All the parts of M. esculenta plant have huge species (ROS) can potentially cause cellular damage. nutritional and therapeutic importance. Fruits are used Oxidative stress is an important contributor to the for syrup, jams, pickels and preparation for refreshing pathophysiology of a variety of human pathological drinks.[9,10] Kafal is a naturally occurring antioxidant, it is conditions including cardiovascular dysfunction, widely used in folk medicine to treat ailments such as atherosclerosis, inflammation, carcinogenesis, drug cough, chronic bronchitis, ulcers, anaemia, fever, toxicity, reperfusion injury and neurodegenerative diarrhea and ear, nose and throat disorders.[11] M. diseases .[1] Plant contains many phytochemical that are esculenta has been studied for its chemical property and useful sources of natural antioxidant, such as phenolic a number of phytochemicals of differet type has been diterpenes, flavonoids, tannins and phenolic acid.[2,3] isolation and characterization from various parts of Polyphenols, especially flavonoids are generally known plants. Recently reported pharmacognostical and as the antioxidant agent in plant extracts.[4] phytochemical activity of M. esculenta steam.[12,13] However, no systematic phytochemical and biological Myrica esculenta belongs to Family Myricaceae, Plant studies have so far been reported on fruits of the plant. species of this genus are distributed in china, Taiwan, The aim of study, therefore, was evaluation of total Japan, Western Highland of Cameroon, North America, polyphenols, flavonoids and antioxidant activity, Nepal and India.[5-8] Myrica esculenta is an important phytochemistry M. esculenta of fruits by DPPH radical medicinal plant in the sub-tropical Himalayas tracks upto scavenging assay. an altitude of 1200 to 2000m and flowering of M. esculenta is February-April and fruiting May-June, distributed to thoughtout commonly known as box berry, fruits are sweet and local name is Kafal or Kaphal. Kafal www.wjpmr.com │ Vol 7, Issue 2, 2021. │ ISO 9001:2015 Certified Journal │ 186 Nishat et al. World Journal of Pharmaceutical and Medical Research 2. MATERIALS AND METHODS replicated three times. It is based on the results of qualitative phytochemical screening, petroleum ether 2.1 Chemicals and Reagents (60-800), chloroform, ethylacetate, acetone and methanol All the chemicals and reagents used for the analytical extracts of Myrica esculenta fruits were selected for works were of laboratory grade and refer to Merck and evaluation of TPC and TFC and antioxidant activity. SD fine-chem limited. DPPH (1, 1-diphenyl-2- picrylhydrazyl), Folin-Ciocalteu reagent, sodium 2.5. Evaluation of total phenolic content (TPC) carbonate, aluminium chloride were all purchased from Total polyphenolic content (TPC) based on the results of Merck, USA. Specified detecting reagents like qualitative phytochemical analysis, petroleum ether (60- Dragendorff’s reagent, FeCl (Ferric choride), Ehrlich 3 800), chloroform, ethylacetate, acetone and methanol reagent, Ninhydrine etc. (for qualitative analysis) extracts were selected for estimation of total phenolic depends upon the nature and class of chemical content. The concentration of phenolics in the extracts constituents. Some reagents were freshly prepared for was determined using spectrophotometic method.[21] qualitative phytochemical screening of fruits extracts. Most active extract solution of concentration of 1mg/ml Some of the reagents were prepared during the was used in the analysis. Methanol solution of the extract experiments while some of the readymade available was used the analysis. Mixture was prepared by mixing reagents were used for qualitative detection. 0.5ml of methanol solution of extract; 2.5ml of 10% Spectrophotometer UV/Vis double beam (Chemito 1700, FolinCiocalteu reagent dissolved in the water and 2.5ml Thermo Fisher) was used for spectrophotometric 7.5% Na CO3. Blank test tube prepared containing 0.5ml determination of TPC (Total phenolic content), TFC 2 methanol, 2.5ml 10% Folin-Ciocalteu reagent dissolved (Total flavonoid content) and free radical scavenging in water and 2.5ml of 7.5% of Na CO . The solution was activity. 2 3 thereafter incubated at room temperature for 45min. The absorbance was determined using spectrophotometer at 2.2. Plant materials λ = 765nm. The sample was prepared in triplicate for Fresh fruits of Myrica esculenta diseases free were max each analysis and the mean value of absorbance was collected from outskirts of Chail Chowk, Mandi, obtained. The same procedure was repeated for the Himachal Pradesh (H.P.), India. The fruits were standard solution of Gallic acid or calibration line was identified and authenticated by Systematic Botany made. Measured absorbance, the concentration of Section of Department of Botany, FRI. A voucher phenolics was read (mg/ml) from the calibration line; specimen has been preserved in the Chemistry & then the content of phenolics in extracts was expressed in Bioprospecting Division, FRI for future reference. The terms of Gallic acid equivalent (mg of GAE/100mg of collected fruits were cleaned properly under running tap extract). Over all the experiments were replicated thrice water to make them free from dust and then dried in and the data were presented as mean values ± standard shade at room temperature (250C). Dried fruits were deviation.[22, 23] stored in airtight bags at room temperature till future use. 2.6. Evaluation of total flavonoid content 2.3. Preparation of extracts Deviation The flavonoid content of chloroform, ethyl Dried Myrica esculenta fruits berries were successively acetate, acetone and methanol extracts was measured extracted with solvents, petroleum ether (60-800), based on methods [24]. Briefly, 0.5ml of sample was chloroform, ethylacetate, acetone, and methanol. The mixed with 1.5ml of methanol and then, 0.1ml of 10% exhaustive extraction was done by shaking the contents aluminium chloride was added, following by 0.1ml of at a regular interval of time till discolored solvents. potassium acetate and 2.8ml of distilled water. The Extracts so obtained were separately distilled under mixture was incubated at room temperature for 30 min. reduced pressure to obtained solvent free extracts and the The absorbance was measured by a spectrophotometer at extractive values were determined on dry weight basis λ = 415nm. The samples were prepared in triplicate with respect to the initial weight of dried and box berry max for each analysis and the mean value of absorbance was pulp taken. The resultant crude extracts were transferred obtained. The same procedure was repeated for the into airtight sample bottles and kept at 4 0C until they standard solution of quercetin (QE) and the calibration were used.[14] line was construed. The total flavonoids content in extracts was expressed in quercetin equivalent (mg of 2.4. Analysis of Phytochemical screening QE/100mg of extract). The standard curve was prepared 1 gram of all extracts fraction was dissolved
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