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2.2.1. Basella Alba A Comparative Study of Cholinesterase Inhibitory, Thrombolytic and Antioxidant Activities of Three Medicinal Plants (Basella alba, Blumea balsamifera, and Curculigo orchioides) Available in Bangladesh for the Treatment of Neurodegenerative Disorders and Clotting Disorders A research paper is submitted to the Department of Pharmacy, East West University in conformity with the requirements for the degree of Bachelor of Pharmacy. Submitted by Farzana Khan Sristy Id: 2013-1-70-080 Department of Pharmacy East West University Submitted to Tirtha Nandi Lecturer Department of Pharmacy East West University East West University Dedicated To My Beloved Parents Without Whom I Could Not Be Here…… I Certificate by the Chairperson This is to certify that the thesis entitled “A Comparative Study of Cholinesterase Inhibitory, Thrombolytic and Antioxidant Activities of Three Medicinal Plant (Basella alba, Blumea balsamifera, and Curculigo orchioides) Available in Bangladesh for the Treatment of Neurodegenerative Disorders and Clotting Disorders” submitted to the Department of Pharmacy, East West University for the partial fulfillment of the requirement for the award of the degree Bachelor of Pharmacy, was carried out by Farzana Khan Sristy, Id: 2013-1-70-080, during the period 2016 of her research in the Department of Pharmacy, East West University. ____________________________ Dr. Shamsun Nahar Khan Associate Professor & Chairperson Department of Pharmacy East West University, Dhaka II Certificate by the Supervisor This is to certify that the thesis entitled “A Comparative Study of Cholinesterase Inhibitory, Thrombolytic and Antioxidant Activities of Three Medicinal Plant (Basella alba, Blumea balsamifera, and Curculigo orchioides) Available in Bangladesh for the Treatment of Neurodegenerative Disorders and Clotting Disorders” submitted to the Department of Pharmacy, East West University for the partial fulfillment of the requirement for the award of the degree Bachelor of Pharmacy, was carried out by Farzana Khan Sristy, Id: 2013-1-70-080, during the period 2016 of her research in the Department of Pharmacy, East West University, under the supervision and guidance of me. The thesis has not formed the basis for the award of any other degree/diploma/fellowship or other similar title to any candidate of any university. __________________________ Tirtha Nandi Lecturer Department of Pharmacy East West University, Dhaka. III Certificate by the Co-Supervisor This is to certify that the thesis entitled “A Comparative Study of Cholinesterase Inhibitory, Thrombolytic and Antioxidant Activities of Three Medicinal Plant (Basella alba, Blumea balsamifera, and Curculigo orchioides) Available in Bangladesh for the Treatment of Neurodegenerative Disorders and Clotting Disorders” submitted to the Department of Pharmacy, East West University for the partial fulfillment of the requirement for the award of the degree Bachelor of Pharmacy, was carried out by Farzana Khan Sristy, Id: 2013-1-70-080, during the period 2016 of her research in the Department of Pharmacy, East West University, under the supervision and guidance of me. The thesis has not formed the basis for the award of any other degree/diploma/fellowship or other similar title to any candidate of any university. __________________________ Kushal Biswas Lecturer Department of Pharmacy East West University, Dhaka. IV Declaration by the Candidate I, Farzana Khan Sristy, hereby declare that the dissertation entitled “A Comparative Study of Cholinesterase Inhibitory, Thrombolytic and Antioxidant Activities of Three Medicinal Plant (Basella alba, Blumea balsamifera, and Curculigo orchioides) Available in Bangladesh for the Treatment of Neurodegenerative Disorders and Clotting Disorders” submitted by me to the Department of Pharmacy, East West University and in the partial fulfillment of the requirement for the award of the degree Bachelor of Pharmacy, work carried out by me during the period 2016 of my research in the Department of Pharmacy, East West University, under the supervision and guidance of Tirtha Nandi, Lecturer, Department of Pharmacy, East West University. The thesis paper has not formed the basis for the award of any other degree/diploma/fellowship or other similar title to any candidate of any university. Farzana Khan Sristy Id: 2013-1-70-080 Department of Pharmacy East West University, Dhaka. V ACKNOWLEDGEMENT At first, I am grateful to God for the good health and wellbeing that were necessary to complete this research. I would like to express my deepest gratitude to my research supervisor, Tirtha Nandi, Lecture, Department of Pharmacy, East West University and co-supervisor, Kushal Biswas, Lecturer, Department of Pharmacy, East West University, who had been always optimistic and full of passion and ideas. Their generous advice, constant supervision, intense support, enthusiastic encouragements and reminders during the research work not only helped shape this study but also molded me into being a better researcher. Their in-depth thinking, motivation, timely advice and encouragement have made it possible for me to complete this research. Secondly, I am also indebted to the Department of Pharmacy, East West University. I am very proud to the part of this institute. To me it seems like second home. This institute is giving me an opportunity to learn about my future goals, to learn how to show respect to the pharmacy profession. I would like to show my gratitude to the Dr. Shamsun Nahar Khan, Associate Professor and Chairperson of Pharmacy Department; Dr. Sufia Islam, Associate Professor of Pharmacy Depertment; JMA Hannan, PhD(UK), Professor of Department of Pharmacy, to the faculties who are teaching over the last four years to make us ready for the noble profession by becoming a pharmacist. Third, I am thanking our respected lab officers Sipra Biswas, Ruchira Chakrabarty, Ajoy Roy, Sujit Kumar and Shofiqul Islam for helping me by providing equipments and materials as well as their guidance I needed to fulfill the work. Forth, my special thanks Taposhi Sultana Mou, Saiyara Hossain Reevu, Shanjida Alam Rika, and all of my friends, who helped me to conduct the research by being very co- operative to be the part of my study. Because of their tremendous support I could finish the work on time. I also, would like to help my fellow classmates, friends for their continuous support in my stay in this institute. Finally, I am immensely grateful to my beloved parents, Md. Alauddin Khan and Sultana Nasrin for their love and faith in me, especially for their unconditional love in my life. It VI is my parents who made me, who I am now! I also would like to express my heartfelt love to my family for their continuous support and love. Farzana Khan Sristy Id: 2013-1-70-080 Department of Pharmacy East West University, Dhaka. VII Table of Content Chapter Name Content Page number Abstract 1 Chapter 1: Introduction 1.1 Introduction 2 1.2 Disease Progration 3 Signs and Symptoms of 1.3 7 Alzheimer’s Disease 1.4 Stages of Alzheimer’s Disease 8 Current Trend of Alzheimer’s 1.5 10 Disease Treatment Hypothesis of 1.6 12 Alzheimer’s Disease Chapter 2: Plant and Literature Review Plant used for the treatment of 2.1 17 Alzheimer’s disease 2.2 Plants of Evaluation 21 2.2.1 Basella alba 21 2.2.2 Blumea balsamifera 23 2.2.3 Curculigo orchioides 26 2.3 Literature review 31 Pharmacological 2.3.1 Investigational Reviews of 31 Basella alba Pharmacological 2.3.2 Investigational Reviews of 33 Blumea balsamifera Pharmacological 2.3.3 Investigational Reviews of 36 Curculigo orchioides Chapter 3: Materials and Methods 3.1 Collection of Plant Materials 38 3.2 Preparation of Plant Material 38 Cold extraction of the plant 3.3 40 materials Determination of Total 3.4 41 Phenolics Test Determination of Total 3.5 43 Flavonoids (TF) Test VIII Chapter Name Content Page number Determination of Total 3.6 45 Flavanol Test DPPH (1, 1-diphenyl-2- 3.7 picrylhydrazyl) Free Radical 47 Scavenging Reducing Power Capacity 3.8 50 Assessment Purification of acetyl 3.9 52 cholinesterase enzyme Preparation of Enzyme Source 3.10 53 (Blood serum) In-Vitro Acetyl 3.11 Cholinesterase Inhibitory 55 Studies In- Vitro Butyryl 3.12 cholinesterase Inhibitory 56 Studies 3.13 Thrombolytic Activity Test 57 Chapter 4: Results Preparation of Crude 4.1 59 Methanolic Extract Determination of Total 4.2 59 Phenolics Determination of total 4.3 flavonoids of crude methanol 61 extracts (CME) Determination of Total 4.4 63 Flavanol DPPH Radical Scavenging 4.5 65 Activity: 4.6 Reducing Power Capacity 67 Acetylcholinesterase 4.7 68 Inhibitory Activity Assay Butyrylcholinestserase 4.8 Inhibitory Activity of 69 Enzymes 4.9 Thrombolytic Activity Test 71 Chapter 5: Discussion and Conclusion 5.1 Discussion 73 5.2 Conclusion 77 Chapter 6: Referencing 6 Referencing 79 IX List of Figure Figure Name Page Anatomical View of Brain in 1.1 2 Alzheimer’s Disease 1.2 Formation of Amyloid Plaque 5 Formation of Neurofibrillary 1.3 5 Tangle Glutamate Mediated 1.4 14 Neurotoxicity 2.1 Ginkgo biloba 17 2.2 Galanthus alpinus 18 2.3 Cannabinoid cannabidiol 19 2.4 Curcuma longa 19 2.5 Curcumin 19 2.6 Huperzine A 20 2.7 Huperzia serrata 20 2.8 Basella alba 22 2.9 Basella alba fruit 22 2.10 Blumea balsamifera 24 2.11 Blumea balsamifera leaf 24 2.12 Curculigo orchioides 27 2.13 C. orchioides Root 27 3.1 Basella alba powder 40 Filtration of Basella alba 3.2 41 extract 3.3 Filtrate of Basella alba 41 3.4 Total Phenolic Test 44 3.5 Total Flavonoid Test 46 Mechanism of DPPH˙ with 3.6 an antioxidant having 49 transferable hydrogen radical 3.7 DPPH test 50 X Figure Name Page Reducing Power Capacity 3.8 52 Assessment Preparation of Butyryl 3.9 cholinesterase enzyme 55 3.10 Thrombolytic activity Test 59 Standard curve of Gallic 4.1 acid for the determination 61 of total phenolics.
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