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Phytochemical and Biological Investigation of Methanolic Leaf Extract of Atalantia Roxburghiana

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Phytochemical and Biological Investigation of Methanolic Leaf Extract of Atalantia Roxburghiana Phytochemical and Biological Investigation of Methanolic Leaf Extract of Atalantia roxburghiana A project submitted by Armin Tania ID: 13146005 Session: Spring 2013 to The Department of Pharmacy In partial fulfillment of the requirements for the degree of Bachelor of Pharmacy (Hons.) Dhaka, Bangladesh December 2017 This work is dedicated to my parents and my sibling to whom I owe my achievements. Certification statement This is to certify that, this project titled ‘Phytochemical and Biological Investigation of Methanolic Leaf Extract of Atalantia roxburghiana’ submitted for the partial fulfillment of the requirements for the degree of Bachelor of Pharmacy (Hons.) from the Department of Pharmacy, BRAC University constitutes my own work under the supervision of Shejuti Rahman Brishty, Lecturer, Department of Pharmacy, BRAC University and this project is the result of the author’s original research and has not previously been submitted for a degree or diploma in any university. To the best of my knowledge and belief, the project contains no material previously published or written by another person except where due reference is made in the project paper itself. Signed, __________________________________ Countersigned by the Supervisor ____________________________________ i Acknowledgement I would like to begin my gratitude to Allah for the help in the completion of this research and preparation of this paper. I would like to thank my supervisor Shejuti Rahman Brishty, Lecturer, Department of Pharmacy, BRAC University for her continuous provision, guidance and patience since the first day of the project work. As a person, she has continuously inspired and motivated me with her skilled knowledge which made me more passionate about the project when it began. I am really indebted to Professor Dr. Eva Rahman Kabir, Honorable Chairperson, Department of Pharmacy, BRAC University, for her support, encouragement and kind cooperation all through the project. Finally, I would like to thank the laboratory officers and assistants for their continuous guidance and cooperative behavior. Armin Tania ii Abstract The present study investigated the phytochemical and biological properties of Atalantia roxburghiana leaf methanol extract for the first time. The objective was to establish this plant as a potential therapeutic agent in traditional practice of medicine in Bangladesh. The preliminary phytochemical screening of the crude methanolic extract exhibited the presence of alkaloids, carbohydrates, flavonoids, phenols, phytosterols, tannins, glycosides and saponin. DPPH radical scavenging effect, total flavonoid and phenolic contents and antioxidant capacities were studied through established protocols and the leaf extract showed significant antioxidant potential through these experimentations. Moderate cytotoxic effect was observed in brine shrimp lethality bioassay. The antimicrobial activity was evaluated by disc diffusion method where the plant extract showed maximum activity against the bacteria Escherichia coli & Vibrio cholera. Evaluation of thrombolytic activity using Streptokinase further indicated very significant thrombolytic properties of the methanolic extract. The present data constitutes a preliminary investigation of the plant which might be valuable for performing research and developing new drugs from natural sources in order to treat different kinds of illnesses in our country as well as in other parts of the world. iii Table of contents Certification statement i Acknowledgement ii Abstract iii Table of contents iv List of tables viii List of figures ix Abbreviations x Chapter-1: Introduction 1.1 General information ................................................................................................................ 1 1.2 Relationship of the phytotherapy with the medicinal plants ................................................ 1 1.3 Research of traditional drug in Bangladesh .......................................................................... 2 1.4 Name of the Plant.................................................................................................................... 6 1.5 Information about plant .......................................................................................................... 6 1.5.1 General information about Atalantia roxburghiana .......................................................... 6 1.5.2 The plant Family: Rutaceae................................................................................................. 8 1.5.3 The plant Genus: Atalantia ................................................................................................. 8 1.5.4 Some reported species ......................................................................................................... 8 1.5.5 Taxonomic hierarchy ........................................................................................................... 9 1.5.6 Chemical constituents of Atalantia roxburghiana ............................................................. 9 1.5.7 Edibility of Atalantia roxburghiana ................................................................................. 10 Chapter-2: Methodology 2.1 Chemical investigation of the experimental plants ............................................................. 12 2.2 Collection and preparation of the plant material ................................................................. 12 2.3 Extraction of the plant material ............................................................................................ 12 2.4 Phytochemical screening ...................................................................................................... 13 2.4.1 Detection of alkaloid.......................................................................................................... 13 2.4.1.1 Hager's test ...................................................................................................................... 14 iv 2.4.1.2 Mayer’s test ..................................................................................................................... 14 2.4.1.3 Wagner’s test................................................................................................................... 14 2.4.2 Detection of carbohydrates................................................................................................ 14 2.4.2.1 Molisch’s test .................................................................................................................. 14 2.4.2.2 Fehling’s test ................................................................................................................... 14 2.4.3 Detection of flavonoids ..................................................................................................... 15 2.4.3.1 Lead acetate test .............................................................................................................. 15 2.4.3.2 Zinc ribbon test ............................................................................................................... 15 2.4.4 Detection of phenols or phenolic compound ................................................................... 15 2.4.5 Detection of phytosterols................................................................................................... 15 2.4.6 Detection of steroids .......................................................................................................... 15 2.4.7 Detection of tannin............................................................................................................. 16 2.4.7.1 Lead acetate test .............................................................................................................. 16 2.4.7.2 Potassium dichromate test .............................................................................................. 16 2.4.8 Detection of resin ............................................................................................................... 16 2.4.9 Detection of glycosides ..................................................................................................... 16 2.5 Evaluation of antioxidant activity ........................................................................................ 17 2.5.1 Determination of Total Phenolic Content (TPC) ............................................................. 17 2.5.1.1 Principle........................................................................................................................... 18 2.5.1.2 Materials and methods.................................................................................................... 18 2.5.1.3 Standard curve preparation ............................................................................................ 19 2.5.1.4 Sample preparation ......................................................................................................... 20 2.5.1.5 Total phenolic compound analysis ................................................................................ 20 2.5.2 Antioxidant activity: DPPH assay .................................................................................... 21 2.5.2.1 Principle........................................................................................................................... 21 2.5.2.2 Materials and methods.................................................................................................... 21 2.5.2.3 Control preparation for antioxidant activity measurement .........................................
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