Phytochemical Screening & in vitro Antioxidant and Thrombolytic Activities of Aphanamixis polystachya (Wall.) Parker Leaf Extracts Submitted by: Ferdous Ara ID: 2010-3-79-030 Research Supervisor: Mr. Apurba Sarker Apu, Senior Lecturer. Department of Pharmacy East West University A thesis report, submitted to the Department of Pharmacy, East West University, in partial fulfillment of the requirements for the degree of Masters of Clinical Pharmacy and Molecular Pharmacology RESEARCH REPORT ON “Phytochemical Screening and in vitro Antioxidant and Thrombolytic Activities of Aphanamixis polystachya (Wall.) Parker Leaf Extracts” DECLARATION BY THE CANDIDATE It is my pleasure to submit the Research Report. The report is prepared on “Phytochemical Screening and in vitro Antioxidant and Thrombolytic Activities of Aphanamixis polystachya (Wall.) Parker Leaf Extracts” which was carried out by me under the guidance of Mr. Apurba Sarker Apu, Senior lecturer, Department of Pharmacy, East West University, Dhaka. I have given my best effort in preparing this report and to make it a worthy one. I have enjoyed working on the report and gained some experience. ……………. Ferdous Ara ID: 2010-3-79-030 Department of Pharmacy East West University, Dhaka. CERTIFICATE BY THE INVIGILATOR This is to certify that the dissertation, entitled “Phytochemical Screening and in vitro Antioxidant and Thrombolytic Activities of Aphanamixis polystachya (Wall.) Parker Leaf Extracts” is a bonafide research work done by Ferdous Ara, in partial fulfillment of the requirement for the Degree of Masters of Clinical Pharmacy and Molecular Pharmacology. ………………………. Mr. Apurba Sarker Apu Senior Lecturer Department of Pharmacy East West University, Dhaka. ENDORSEMENT BY THE CHAIRPERSON This is to certify that the dissertation, entitled “Phytochemical Screening and in vitro Antioxidant and Thrombolytic Activities of Aphanamixis polystachya (Wall.) Parker Leaf Extracts” is a bonafide research work done by Ferdous Ara, under the guidance of Mr. Apurba Sarker Apu, Senior Lecturer, Department of Pharmacy, East West University, Dhaka. ………………. Dr. Sufia Islam Chairperson and Associate Professor Department of Pharmacy East West University, Dhaka. Acknowledgement Above all, I express my gratitude to Almighty Allah for giving me the strength, energy and patients to carry out this research work. I would like to express my gratitude and admiration to Mr. Apurba Sarker Apu, Senior Lecturer, Department of Pharmacy, East West University, for his suggestion, careful guidance, sincere help, constructive criticism and valuable time without which I would not have been able to complete this work. He had been very enthusiastic and supportive in my research. I would like to extend my cordial gratitude to Dr. Sufia Islam, Chairperson and Associate Professor of the Department of Pharmacy, East West University, for her constant inspiration and whole hearted cooperation. I would also like to take the opportunity to express my whole hearted gratitude to my fellow researchers friends, near and dear ones who offered encouragement, information, inspiration and assistance during the period of constructing the research report. I am thankful to the laboratory instructors for their kind support during the laboratory work. I express my sincere gratitude to my caring parents for their support, inspiration and guiding me all through my life, including that for my research project. I am very grateful to my sisters who encouraged me enormously. This Research paper is dedicated to My beloved parents and sisters TABLE OF CONTENTS CHAPTER 1: INTRODUCTION List of Contents Page No. Introduction 1-3 Plant Profile 3 Common name 3 Botanical name 3 Synonyms 3 Trade names 4 Scientific classification 4 Distributional Range 4 Native distribution 5 Botanical Descriptions of A. polystachya 5 Habit 5 Foliage 6 Plant shape 6 Trunk & bark 6 Branches and branchlets 6 Leaves 6-7 Inflorescence / Flower 7 Flowering 8 Fruit and seed 8 Type of stem 8 Wood 8 Roots 9 Plant utilities 9 Season 9 Ecology 9 Major Threat 9 Uses of A. polystachya 9 1. Medicinal uses 9 2. Common uses 10 Solvent System 10 1. n-hexane 10 Applications 10-11 2. Ethyl acetate 11 Applications 11-12 3. Methanol 12 Applications 12-13 Phytochemical Screening 13-17 Antioxidants 17 Free radical formation by oxygen 17-18 Problem of free radical 19 DPPH free radical scavenging assay 19-20 Nitric oxide scavenging 20-21 Estimation of total phenolic content 21-22 Thrombolytic Activity 22-25 Purpose of the Present Study 25 CHAPTER 2: LITERATURE REVIEW List of contents Page No. 2.1 Phytochemical Studies 26-31 2.2 Pharmacological Studies 31-39 CHAPTER 3: MATERIALS & METHODS List of Contents Page No. Collection and Identification of Plant Material 40 Materials 41 Method 41 Extraction of plant material 41-42 Condensation of the leaf extracts 42-43 Screening of Phytochemical Constituents 44 Test for alkaloids: Hager's test 44 Materials 44 Method 44 Preparation of 2% H SO solution 44 2 4 Preparation of Hager´s reagent 44 Procedure 44-45 Test for flavonoids: Ammonia test (modified) 45 Materials 45 Method 45 Preparation of 5% ammonia solution 45-46 Procedure 46 Test for steroids: Salkowski test 46 Materials 46 Method 47 Procedure 47 Test for terpenoids: Salkowski test (modified) 47 Materials 47 Method 47 Procedure 48 Test for carbohydrates: Fehling's (Reducing sugar) test (modified) 48 Materials 48 Method 48