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Screening of Phyto-chemical compounds from hydro-ethanolic and ethanolic leaf and bark extracts of Terminalia arjuna and Syzygium cumini
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HARMONY IN THE NATURE AND EXISTENCE - Unit-IV
HARMONY IN THE NATURE AND EXISTENCE - Unit-IV
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Porous Ceramics seminar and technical writing
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Screening of Phyto-chemical compounds from hydro-ethanolic and ethanolic leaf and bark extracts of Terminalia arjuna and Syzygium cumini
1.
International Research Journal
of Engineering and Technology (IRJET) e-ISSN: 2395 -0056 Volume: 04 Issue: 06 | June -2017 www.irjet.net p-ISSN: 2395-0072 © 2017, IRJET | Impact Factor value: 5.181 | ISO 9001:2008 Certified Journal | Page 1934 Screening of Phyto-chemical compounds from hydro-ethanolic and ethanolic leaf and bark extracts of Terminalia arjuna and Syzygium cumini Dr. Swaati Sharma , Priya Vishnoi Department of Agriculture, Food and Biotechnology, Jayoti Vidyapeeth Women’s University, Jaipur, Rajasthan, India. ---------------------------------------------------------------------------***--------------------------------------------------------------------------- Abstract: Medicinal plants have many phyto-chemicals or secondary metabolites which are used to treat many human diseases (oxidative stress diseases and neurodegenerative diseases). Neurodegenerative diseases include stroke, Parkin-son’s disease and Alzheimer’s disease and oxidative stress diseases such as inflammatory bowel diseases, retinal ischemia and cardiovascular diseases. In this study, different parts of Terminalia arjuna and Syzygium cumini plants (mainly leaves and bark) are used for screening of bioactive compounds. Plant parts are extracted with ethanolic and hydro-ethanolic (50%) solvents by soxhlet apparatus extraction method and screened for the presence of secondary metabolites namely alkaloids, flavonoids, rasins, saponins, steroids, tannins, glycosides, phenol, terpenoids and fatty acids. In this research work, We found that selected plants are good source of various bioactive compounds. However, both leaves and bark of Terminalia arjuna contain high amount of phyto-chemicals than Syzygium cumini. From this study, We concluded that both plants are good source of secondary metabolites and can be used as medicinal source for the treatment of various diseases. KEYWORDS: Terminalia arjuna, Syzygium cumini, secondary metabolites, phyto-chemicals, bioactive compounds. 1. INTRODUCTION: Since ancient times, secondary metabolites have ability to prevent chronic diseases far away their role as dietary antioxidants (1). According to World Health Organisation (WHO), medicinal plants can be good source to obtain variety of drugs (2). Generally herbal drugs are less expensive, safe, and easily available and rarely have side effects. These are widely used in human therapy, veterinary, scientific research and agriculture etc. A large number of phyto-chemicals belong to various chemical classes have prohibitory effects on all types of microorganism invitro. Medicinal plants contain many bioactive compounds and are concentrated in the endangered parts of the plants such as leaves, bark, flowers, fruit and skin, used for various medicinal purposes (3,4, 5). Secondary metabolites have their role in several health benefits and provide a source of medicine since historic times. Traditional plants are helpful in world’s pharmaceuticals and serve as starting material in drug development. It has been reported that these natural secondary metabolites may reduce the risk of cardiovascular diseases, oxidative stress diseases and neurodegenerative diseases (1). The plants chosen for this study are Terminelia arjuna (T.arjuna) and Syzygium cumini (S. cumini). T.arjuna is large sized tree to 30 m tall, belongs to Combretaceae family. This large an evergreen tree appeared in hot season (February to April), found in South Asian region having wide spectrum of biological activity. The bark of T. arjuna have been reported for anti-dysentric, antipyretic, astringent, cardiotonic, lithotriptic, anticoagulant, hypolipidemi, antimicrobial (6) and antiuremic (7) activities. Many useful phytochemicals have been isolated from T.arjuna including terpenoids for cardiovascular properties, tannins and flavonoids for its antimicrobial and anticancer properties etc (8). On the other hand, S.cumini is commonly known as jambolan or java plum, belongs to family Myrtaceae. Plants of this family are rich in volatile oils which are used in medicines (9). Fruits of this family are used as edibles and as traditional medicines in divergent ethno-botanical practices throughout the tropical and subtropical world (10). The fruits are berries (edible), 1.5 to 3.5 centimeters long, dark purple and nearby black color contain single large seed (11, 12). The fruit is sweet, mildly sour and astringent in taste and contains raffinose, glucose, fructose (13), citric acid, mallic acid (14), gallic acid, anthicyanins (15).
2.
International Research Journal
of Engineering and Technology (IRJET) e-ISSN: 2395 -0056 Volume: 04 Issue: 06 | June -2017 www.irjet.net p-ISSN: 2395-0072 © 2017, IRJET | Impact Factor value: 5.181 | ISO 9001:2008 Certified Journal | Page 1935 2. MATERIALS AND METHODS:- 2.1COLLECTION OF PLANT MATERIAL: The leaves and bark of T.arjuna and S.cumini were procured from the botanical garden of jayoti vidhyapeeth women’s university. 2.2EXTRACTION OF PLANT MATERIAL: Fresh plant materials were washed with running tap water and dried at room temperature. After washing and drying, leaves and barks of the plants were grinded using grinder to make powder. The powder was then subjected to successive extraction with different solvents (50% hydro-ethanol and ethanol) using soxhlet apparatus method. The hydro-ethanolic and ethanolic extracts were prepared by taking 25 grams of each powdered sample and soaked in 250 ml of 50% hydro-ethanol or ethanol solvent system. The collected extracts were stored in calcium carbonate at very low temperature and then taken up for further research work. 2.3PHYTOCHEMICAL ANALYSIS: Chemical test were carried out on 50% hydro-ethanolic and ethanolic extracts of aerial parts of T.arjuna and S.cumini using standard procedures to identify the constituents (16,17,18). Test for Glycosides: For the estimation of glycosides, extract was diluted with solvent by which extract was extracted from plant materials. 2ml of diluted plant extract was mixed with 2ml of chloroform and 2ml of sulphuric acid then shaken well. Reddish brown color indicated the presence of steroidal ring (glycone portion of glycosides). Test for Fatty acids: 0.5gm of plant extract was mixed with 5ml of ether and then extract was evaporated on filter paper and dried the filter paper. The transparency appeared on filter paper indicated the presence of fatty acids. Test for Flavonoids: Alcoholic solution of extract was mixed with few drops of 10% lead acetate. Yellow precipitate indicated the presence of flavonoids. Test for Phenol: Diluted extract was mixed with few drops of ferric chloride solution. Appearance of bluish black color showed the presence of phenol. Test for Resins: 0.5gm of plant extract was diluted with 10 ml of water and then shaken for 5 min. Turbidity formed in the solution indicated the presence of resins. Test for Saponins: Diluted 5ml extract was stirred with 5ml of distilled water and then warmed in a test tube. Stable formation of foam indicated the presence of saponins. Test for Steroids: Diluted 2ml of extract was stirred with 2ml of chloroform and 2ml of conc. Sulphuric acid. Red color produced in the lower chloroform layer indicated the presence of steroids. Test for Tanins: 2ml of extract was mixed with 2ml of DW and then 2 or 3 drops of ferric chloride was mixed to the solution. Green precipitate indicated the presence of tannins. Test for Terpenoids: To identify the presence of terpenoids, Salkwoski method was used. 0.5 gm plant extract was mixed with 2ml of chloroform and 3ml of conc. Sulphuric acid to form a layer. Reddish brown color indicated the presence of terpenoids.
3.
International Research Journal
of Engineering and Technology (IRJET) e-ISSN: 2395 -0056 Volume: 04 Issue: 06 | June -2017 www.irjet.net p-ISSN: 2395-0072 © 2017, IRJET | Impact Factor value: 5.181 | ISO 9001:2008 Certified Journal | Page 1936 QUALITATIVE ESTIMATION OF PHYTOCHEMICALS:- TABLE NO:- 1 NAME OF PHYTO- CHEMICALS NAME OF MEDICINAL PLANTS TERMINALIA ARJUNA SYZYGIUM CUMINI LEAVES AND BARK EXTRACTED WITH 50% HYDRO-ETHANOL LEAVES AND BARK EXTRACTED WITH ETHNOL LEAVE AND BARK EXTRACTED WITH 50% HYDRO- ETHANOL LEAVES AND BARK EXTRACTED WITH ETHANOL Leaves Bark Leaves Bark Leaves Bark Leaves Bark Saponins ++ +++ ++ ++ ++ ++ +++ ++ Steroids ++ +++ ++ + +++ ++ ++ + Flavonoids +++ ++ +++ ++ ++ +++ +++ ++ Phenols +++ ++ +++ ++ +++ +++ +++ ++ Resins +++ + ++ + +++ +++ + ++ Tanins +++ ++ +++ ++ ++ ++ ++ ++ Terpenoids +++ ++ +++ ++ ++ + ++ ++ Fatty acids +++ - +++ - - - - - Glycosides +++ +++ +++ +++ +++ ++ +++ ++ +++ High concentration, ++ Moderate concentration, + Low concentration, - Absence. 3.RESULT AND DISCUSSION : The qualitative estimation of phyto-chemicals of plants T.arjuna and S. cumini exhibited the presence of various bioactive components such as glycosides, saponins, flavonoids, phenol, resins, tannins, terpenoids, fatty acids. From this study, I found that T.arjuna contains high amount of phyto-chemicals than S.cumini. Different plant parts with different solvents were screened to check the presence of various secondary metabolites. Results have been shown in table no.1. Leaves of T.arjuna extracted with 50% hydro-ethanol contain high (+++) amount of flavonoids, phenols, resins, tannins, terpenoids, fatty acids and glycosides and moderate (++) amount of saponins and steroids, whereas, the bark extracted with the same solvent contains high (+++) amount of saponins, steroids and glycosides, moderate (++) amount of flavonoids, tannins, terpenoids and phenols, low (+) amount of resins and fatty acids are absent(-). In case of leaves extracted with ethanol contain high (+++) flavonoids, phenol, tannins, terpenoids, fatty acids and glycosides, moderate (++) saponins, steroids and resins, whereas, the bark extracted with the same solvent contains high (+++) glycosides, moderate (++) saponins flavonoids, phenols, tannins and terpenoids, low (+) steroids and resins, fatty acids are absent(-). The bark of T.arjuna functions as a diuretic in cirrhosis of liver and gives relief in symptomatic hypertension (19). Its leaves have been reported to have analgesic and anti-inflammatory properties (20). T.arjuna is also known as cardiotonic since historic times (21). The bark of T.arjuna and juice of it is used as antacid by the people live in malkangiri district of orrisa
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International Research Journal
of Engineering and Technology (IRJET) e-ISSN: 2395 -0056 Volume: 04 Issue: 06 | June -2017 www.irjet.net p-ISSN: 2395-0072 © 2017, IRJET | Impact Factor value: 5.181 | ISO 9001:2008 Certified Journal | Page 1937 (22). Powder of bark is boiled with water and inhaled to cure headache and kill bacteria in teeth (23). Bark ash is used to treat snakebite and scorpion sting (21). The bark of T.arjuna is also used to treat fever and high blood pressure in south surguja district of Madhya Pradesh (24). It has been proven that T.arjuna and its species have potential to treat HIV (25). On the other hand, leaves of S.cumini extracted with 50% hydro-ethanol contain high (+++) steroids, phenols, resins and glycosides, moderate (++) flavonoids, tannins, terpenoids and saponins, fatty acids are absent(-), whereas, bark extracted with the same solvent contains high (+++) flavonoids, phenols and resins, moderate (++) saponins, steroids, tannins and glycosides, low (+) terpenoids and fatty acids are absent (-). However, In case of leaves extracted with ethanol contain high (+++) saponins, flavonoids, phenols and glycosides, moderate (++) steroids, tannins and terpenoids, low (+) resins and fatty acids are absent (-), whereas the bark extracted with the same solvent contains moderate (++) saponins, flavonoids, phenols, resins, tannins, terpenoids and glycosides, low (+) steroids and fatty acids are absent (-). The fruits of S.cumini having medicinal properties, used to treat cough, diabetes, inflammation, dysentery and ringworm (10). The fruit (mainly seeds) of S.cumini is also claimed to contain an alkaloid, jambosine, and a glycoside, jambolin or antimellin, which stop the diastatic coversion of starch into sugar (26). They are also rich in flavonoids (a well known antioxidant), which accounts for the scavenging of free radicals (27, 28). Jambolan also have phenolics with antioxidant activity (29) and is rich in protein and calcium. Java plums are also rich in mineral salts, sugar, vitamin C, PP which boosts the beneficial effects of vitamin anthocyanins and flavonoids (30). Different parts of the jambolan have been reported for its antioxidant, anti-inflammatory, neuropsycho-pharmacological, anti-microbial, anti-bacterial, anti-HIV, antileishmanial and antifungal, nitric oxide scavenging, free radical scavenging, anti-diarrheal, antifertility, anorexigenic, gastroprotective and anti-ulcerogenic and radioprotective activities (31). Phyto-chemicals in our diet promote the health (1). Phyto-nutrients are related with the prevention of recognized chronic diseases including cardio vascular diseases (CVDs), cancer, osteoporosis, diabetes and vision diseases that are mainly terrible in Western countries. These diseases are mostly attributed to a high-sugar and high-fat diet, smoking, lack of exercise and other unhealthy lifestyle (32). Interaction of carcinogens or oxidants with DNA causes many types of cancers. The powerful antioxidant ability of phyto-nutrients can reduce this damage (33). Different parts of plant are also used to treat various diseases such as diabetes, blisters in mouth, cancer, colic, diarrhea, digestive complaints, dysentery, piles, pimples and stomachache (34). Phyto-chemicals, found in bark and leaves of T.arjuna and S.cumini, play important roles to treat various diseases. Tannins have the ability to treat sore throat, wound healing, antidiarrhoea and antihaemorrhagic agent (35, 36). Tannins have also been studied for the antimicrobial degradation of dietary proteins of semen (37). Flavonoids can inhibit the initiation, promotion and progression of tumors. Phenols and flavanoids, both are strong water soluble antioxidants (38). These phenolic compounds also have some biological properties such as anti-apoptosis, anti-septics, anti-carcinogen, anti-ageing, anti- inflammation, anti-atherosclerosis, improvement of endothelial function and cardiovascular protection (36). These phenolic compounds also have ability to inhibit angiogenesis and cell proliferation activities (39). Terpenoids have carboxylic acid group which is responsible for the activity of organic extracts (40,16). Glycosides are present in all the hydro-ethanolic and ethanolic extracts of T.arjuna and S.cumini which lower the blood pressure (41). Saponins, found in hydro-ethanolic and ethanolic extracts of both the plants are known to posses reduction in cholesterol level and can manage cardiovascular diseases in humans (37). Saponins also have been used as emulsifying agents and having anti-fungal (42), anti-inflammatory activity (43), coagulating and precipitating property in red blood cells. Steroids also have been found in these medicinal plants, previously reported for its antibacterial activity (44) and their association with such compounds as sex hormones (45). Fatty acids, found in hydro-ethanolic and ethanolic extracts of T.arjuna leaves are previously reported for their anti-fungal and anti- bacterial activities (46). Resins are also present in these traditional medicinal plants, used for the treatment of arthritis, wound healing and have anti-inflammatory, anti-microbial, anti-tumor, anti-hyperlipidemia properties (47). 4. CONCLUSION: From this study, it can be concluded that these species may be used for drug development in pharmaceutical companies. Different parts (leaves and bark) of both plants are good source of phyto-chemicals which have anti-fungal and anti-microbial properties. ACKNOWLEDGEMENT:- The authors are thankful to Dr. Panckaj Garg, Founder & Advisor and Department of Agriculture, Food and Biotechnology, Jayoti Vidyapeeth Women’s University, Jaipur for providing the laboratory and technical facilities used in this study.
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International Research Journal
of Engineering and Technology (IRJET) e-ISSN: 2395 -0056 Volume: 04 Issue: 06 | June -2017 www.irjet.net p-ISSN: 2395-0072 © 2017, IRJET | Impact Factor value: 5.181 | ISO 9001:2008 Certified Journal | Page 1938 REFERENCE: 1. Yongping Bao, Roger Fenwick, Phytochemicals in health and diseases, Marcel Dekker, INC, New York, Basel, 2004. 2.Yadav RN, Agrawala M. Phytochemical analysis of some medicinal plants. Journal of Physiology. 2011;3(12):10–14 3. Mojab F, Kamalinejad M, Ghaderi N, Vanidipour HR. Phytochemicals screening of some species of Iranian plants. Iranian Journal of Pharmaceutical Research. 2003;3:77–82. 4. Parekh J, Chanda S. Antibacterial and phytochemical studies on twelve species of Indian medicinal plants. African Journal Biomedcal Research. 2007;10:175–181. 5. Parekh J, Chanda S. Phytochemicals screening of some plants from western region of India. Plant Arch.2008;8:657–662. 6. Mandal A, Das K, Nandi D K. In vitro bioactivity study of bark extract of Terminalia arjuna on probiotics, commercially available probiotic formulation. International Journal of Phytopharmacology. 2010;1(2):109–113. 7. Das K, Chakraborty PP, Ghosh D, Nandi DK. Protective effect of aqueous extract of Terminalia arjuna against dehydrating induced oxidative stress and uremia in male rat. Iranian Journal of Pharmaceutical Research. 2010;9(2):153-161. 8. Nema R, jain P, Khare S, Pradhan A, Gupta A, Singh D. Antibacterial and antifungal activity of Terminalia arjuna leaves extract with special reference to flavanoids. Basic Research Journal of Medcine and Clinical Science.2012;1(5):63–65. 9. Mahmoud II, Marzouk MS, Moharram FA, El-Gindi MR, Hassan AM. Acylated flavonol glycosides from Eugenia jambolana leaves. Phytochemistry. 2001;58:1239–1244. 10. Reynertson KA, Basile MJ, Kennelly EJ. Antioxidant potential of seven myrtaceous fruits. Ethnobotany Research and Applications. 2005;3:25–35. 11. Gamble JS. The flora of the presidency of Madras. London: Adlard & Son LTD; 1935. 12. Hooker JD. The flora of British India. London: Nabu Press; 1879. p. 499. 13. Srivastava HC. Paper chromatography of fruit juices. Journal of Scientific and Industrial Research. 1953;12B:363–365 14. Lewis YS, Dwarakanath CT, Johar DS. Acids and sugars in Eugenia jambolana. Journal of Scientific and Industrial Research. 1956;15C:280–281 15. Jain MC, Seshadri TR. Anthocyanins of Eugenia jambolana fruits. Indian Journal of Chemistry. 1975;3:20–23. 16. Harbone J.B., Phytochemicals methods. Chapman and Hall Ltd., London, 1973; 49-188. 17. Trease G.E., Evans W.C., Pharmacognosy, 11th edn., Bailliere Tindall, London, 1989; 45-50. 18. Sofowra A., Medicinal plants and Traditional Medicine in Africa, Spectrum Books Ltd., Ibadan, Nigeria, 1993; 191-289. 19. Chatterjee AS. The treatise on indian medicinal plants: Council of scientific and industrial research. New Delhi: Publication and Information Directorate; 1994. 20. Biswas M, Biswas K, Karan TK, Bhattacharya S, Ghosh AK, Haldar PK. Evaluation of analgesic and anti-inflammatory activities of Terminalia arjuna leaf. Journal of Phytology. 2011;3(1):33–38. 21. Yesodharan K and Sujana KA: Ethnomedicinal knowledge among Malamalasar tribe of Parambikulam wildlife sanctuary, Kerala. Indian Journal of Traditional Knowledge, 2007; 6(3): 481-485. 22. Prusti AB and Behera KK: Ethnobotanical Exploration of Malkangiri District of Orissa, India. Ethnobotanical Leaflets, 2007; 1:12-15. 23. Muthu C Ayyanar Raja N and Ignacimuthu S: Medicinal plants used by traditional healers in Kancheepuram District of Tamil Nadu, India. Journal of ethnobilology and Ethnomedicine, 2006; 2:43. 24. Kumar V and Jain SK: A contribution to Ethnobotany of Surguja district in Madhya Pradesh, India. Ethnobotany, 1998; 10: 89-96. 25. Alka Dwevedi, Raman Dwivedi, Yogesh K Sharma, Exploration of Phytochemicals Found in Terminalia sp. and their Antiretroviral Activitiy, Pharmacognosy Reviews. 2016 Jul-Dec; 10(20): 73–83. 26. Morton J. Fruits of warm climates. Florida flair books , Miami: Julia Morton Winterville North Carolina; p. 1987. 27. Ravi K, Ramachandran B, Subramanian S. Protective effect of Eugenia jambolana seed kernel on tissue antioxidants in streptozotocin induced diabetic rats. Biological and Pharmaceutical Bulletin. 2004;27:1212–1217. 28. Ravi K, Ramachandran B, Subramanian S. Effect of Eugenia jambolana seed kernel on antioxidant defense system in streptozotocin induced diabetes in rats. Life Science journal. 2004a;75:2717–2731. 29. Bajpai M, Pande A, Tewari SK, Prakash D. Phenolic contents and antioxidant activity of some food and medicinal plants. International Journal of Food Science and Nutrition. 2005;56:287–291. 30. Council of Scientific and Industrial Research . The wealth of India. New Delhi: Council of Scientific and Industrial Research; 1948. 1976 pp. 31. Sagrawat H, Mann AS, Kharya MD. Pharmacological potential of Eugenia jambolana: a review. Pharmacognosy Magazine. 2006;2:96–104.
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