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Preliminary Phytochemical Screening of Ethanolic and Aqueous Extracts of Leaves of Plumeria Obtusa

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Preliminary Phytochemical Screening of Ethanolic and Aqueous Extracts of Leaves of Plumeria Obtusa The International journal of analytical and experimental modal analysis ISSN NO:0886-9367 Preliminary Phytochemical Screening of Ethanolic and Aqueous Extracts of Leaves of Plumeria Obtusa Shukla Pawandeep1, Saraswat Rohit2, Gupta Arun Kumar3, Patel Rakesh4, Chaturvedi Mohit5 1, 2, 5 Department of Pharmaceutical Sciences, OPJS University, Churu, Rajasthan, India 3 Chameli Devi Institute of Pharmacy, Indore, Madhya Pradesh, India 4 Department of Pharmacy, Dr. A.P.J. Abdul Kalam University, Indore, Madhya Pradesh, India Abstract Preliminary screening of phytochemicals is a valuable step, in the detection of the bioactive principles present in medicinal plants and subsequently may lead to drug discovery and development. In the present study, chief phytoconstituents of leaves of Plumeria Obtusa plant was identified in order to relate their presence with bioactivities of the plants. Screening of selected medicinal plant part extracts was performed for the presence of tannins, flavonoids, terpenoids, saponins, steroids, phlobatannins, carbohydrates, glycosides, coumarins, alkaloids, proteins, emodins, anthraquinones, anthocyanins and leucoanthocyanins using standard methods, which contribute several pharmaceutical applications to humans. Keywords: Plumeria Obtusa, Phytochemical screening, phytoconstituents, Medicinal plants, Leaves. (Corresponding author: [email protected]) Introduction Medicinal plants besides therapeutic agents are also a big source of information for a wide variety of chemical constituents which could be developed as drugs with precise selectivity. These are the reservoirs of potentially useful chemical compounds which could serve as newer leads and clues for modern drug design [1]. Knowledge of the chemical constituents of plants is desirable because such information will be of value for the synthesis of complex chemical substances. The study of plants continues principally for the discovery of novel secondary metabolites which possess many pharmacological properties. Owing to the significance in the above context, such preliminary phytochemical screening of plants is the need of the hour in order to discover and develop novel therapeutic agents with improved efficacy. Numerous research groups have also reported such studies throughout the world [2-6]. Thus, the present study deals with the screening based on phytochemical tests of ethanolic and aquesous extracts of leaves of Plumeria Obtusa. Volume XII, Issue VI, June/2020 Page No: 1336 The International journal of analytical and experimental modal analysis ISSN NO:0886-9367 Plumeria obtusa is native to the Greater Antilles, Florida, northern Central America and southern Mexico. Cultivation is common in heater parts of the world, including Southeast Asia [7]. In the Caribbean Islands, people use the bark as a diuretic. They also use the latex to stimulate purging [8, 9]. Plumeria obtusa (Synonyms: Plumeria apiculata Urb., Plumeria bahamensis Urb., Plumeria barahonensis Urb., Plumeria beatensis Urb., Plumeria bicolor Seem., Plumeria cayensis Urb., Plumeria clusioides Griseb., Plumeria clusioides Var. parviflora (Griseb.) M.Gómez, Plumeria confusa Britton, Plumeria cubensis Urb., Plumeria cuneifolia Helwig, Plumeria dictyophylla Urb., Plumeria ekmanii Urb., Plumeria emarginata Griseb., Plumeria estrellensis Urb., Plumeria inaguensis Britton, Plumeria jamaicensis Britton, Plumeria krugii Urb., Plumeria marchii Urb., Plumeria montana Britton & P.Wilson, Plumeria nipensis Britton, Plumeria nivea Mill., Plumeria obtusa var. laevis Griseb., Plumeria obtusa var. parviflora Griseb., Plumeria ostenfeldii Urb., Plumeria parvifolia Donn, Plumeria portoricensis Urb., Plumeria tenorei Gazparr., Plumeria venosa Britton, Plumeria versicolor Dehnh., is a plant with comman names Bengali: Kathgolop, Hindi: Golachin, Champa, Golenchi, Kannada: Sampige, Konkani: Champe, Manipuri: Khagi-leihao, Marathi: Chafa, having a place with family Apocynaceae [8-17 ]. Materials and Methods Collection and identification of plant The leaves of Plumeria obtusa was collected in the month of July 2016 from local areas of Indore (M.P.) and were identified and authenticated by Dr. S.N. Dwivedi, Prof. and Head, Department of Botany, Janata PG College, A.P.S. University, Rewa, M.P., India. what's more, kept in our Laboratory, Voucher example No. J/BOT/H-139. Preparation of plant extracts The dried powder of leaves of Plumeria obtusa was separated with Ethanol (95%) in a soxhlet apparatus. Aqueous extract was set up by cold maceration process by utilizing separate amount of powder. The solvents were evacuated by refining under diminished weight and the subsequent semisolid mass was vacuum dried utilizing rotary flash evaporator [18]. Phytochemical screening Phytochemical screening was performed to identify phytochemicals in the Ethanolic and Aqueous extracts of Plumeria obtusa leaves exposed to following test to decide the presence of phytoconstituents. Volume XII, Issue VI, June/2020 Page No: 1337 The International journal of analytical and experimental modal analysis ISSN NO:0886-9367 Tests for carbohydrates and glycosides A small quantity of the extracts was dissolved individually in 4 ml of distilled water and filtered. The filtrate was exposed to Molisch’s test to find out the presence of Carbohydrates. Molisch’s test Filtrate was treated with 2-3 drops of 1% alcoholic - napthol arrangement and 2 ml of Conc. sulphuric acid was included at the edges of the test tube. Appearance of dark colored ring at the intersection of two fluids demonstrates the presence of carbohydrates. Another bit of the extract was hydrolyzed with hydrochloric acid for couple of hours on a water bath and the hydrolysate was exposed to Legal's and Borntrager's test to distinguish the presence of various glycosides. Legal’s test To the hydrolysate 1 ml of pyridine and couple of drops of sodium nitropruside solutions were added and then it was made alkaline with sodium hydroxide solution. Appearance of pink to red colour indicates the presence of glycosides. Borntrager’s test Hydrolysate was treated with chloroform and then the chloroform layer was removed. To this, equal quantity of dilute ammonia solution was added. Ammonia layer acquires pink color, showing the presence of glycosides. Test for alkaloids A small section of the solvent free pet ether, alcohol extracts were stirred individually with few drops of dil. hydrochloric acid and filtered. The filtrate was tested with number of reagents for the presence of alkaloids. The reagents are following: Dragondroff’s reagent : Reddish brown ppt Wagner’s reagent : Reddish brown ppt Mayer’s reagent : Cream color ppt Hager’s reagent : Yellow ppt Volume XII, Issue VI, June/2020 Page No: 1338 The International journal of analytical and experimental modal analysis ISSN NO:0886-9367 Test for proteins and free amino acids Small quantities of the extracts were dissolved in few ml of water and treated with following reagents and observed. Million’s reagent : Appearance of red color shows the Presence of protein and free amino acid. Ninhydrin reagent : Appearance of purple color shows the presence of proteins and free amino acids Biuret’s test : Equal volumes of 5% sodium hydroxide solution & 1% copper sulphate solution was added. Appearance of pink or purple color shows the presence of proteins and amino acids. Test for phenolic compounds and tannins Small quantities of the extracts were taken individually in water and test for the presence of phenol compounds and tannins was carried out with the following reagents. 1. Dil. Ferric chloride solution (5%): Violet color. 2. 1% solution of gelatin containing 10% sodium chloride: White ppt 3. 10% lead acetate solution: White ppt Test for flavonoids With aqueous sodium hydroxide solution Blue to violet colour (anthocyanins) yellow colour (flavones), yellow to orange (flavonones) With con. sulphuric acid Yellow orange colour (anthocyanins) yellow to orange colour (flavones) orange to crimson (flavonones) Shinoda’s test Small quantities of the extract were dissolved in alcohol, to them piece of magnesium followed by Con: hydrochloric acid drop wise added and heated. Appearance of magenta color shows the presence of flavonoids. Volume XII, Issue VI, June/2020 Page No: 1339 The International journal of analytical and experimental modal analysis ISSN NO:0886-9367 Tests for fixed oils and fats Spot test Small quantities of various extract were separately pressed between two filter papers. Appearance of oil stain on the paper indicates the presence of fixed oil. Few drops of 0.5 N alcoholic potassium hydroxide were added to a small quantity of various extracts along with a drop of phenopthlein, the mixture was heated on a water bath for 1-2 hours, soap formation or partial neutralization of alkali indicates the presence of fixed oils and fats. Tests for steroids and triterpenoids Libermann-Burchard test Treat the extract with couple of drops of acetic anhydride, boil and cool. Then add con. sulphuric acid from the side of test tube, dark brown ring is formed at the junction two layers and upper layer turns green which shows presence of steroids and formation of deep red colour indicates presence of triterpinoid. Salkowski test Treat the extract with few drop of con. Sulphuric acid, red colour at lower layer indicates presence of steroids and formation of yellow coloured lower layer indicates presence of triterpenoids. Test for mucilages and gums Small quantities of extract were added separately to 25 ml of absolute alcohol with constant
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