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University of Gondar College of Natural And UNIVERSITY OF GONDAR COLLEGE OF NATURAL AND COMPUTATIONAL SCIENCE DEPARTMENT OF CHEMISTRY DETERMINATION OF ESSENTIAL METAL, POLYPHENOL CONTENT, ANTIOXIDANT AND ANTIBACTERIAL ACTIVITIES OF METHANOLIC ROOT EXTRACTS OF CROTON MACROSTACHYUS AND PHYTOLACCA DODECANDRA A THESIS SUBMITTED TO DEPARTMENT OF CHEMISTRY IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN CHEMISTRY BY: ABEBE TSEGA ADVISOR: DESSIE TIBEBE (PhD, ASSOCIATE PROFESSOR) CO-ADVISOR: MR. MULUGETA LEGESSE (M.Sc., ASSISTANT PROFESSOR) OCTOBER, 2020 GONDAR, ETHIOPIA i UNIVERSITY OF GONDAR COLLEGE OF NATURAL AND COMPUTATIONAL SCIENCE DEPARTMENT OF CHEMISTRY DETERMINATION OF ESSENTIAL METAL, POLYPHENOL CONTENT, ANTIOXIDANT AND ANTIBACTERIAL ACTIVITIES OF METHANOLIC ROOT EXTRACTS OF CROTON MACROSTACHYUS AND PHYTOLACCA DODECANDRA BY: ABEBE TSEGA APPROVED BY THE EXAMINING BOARD: Name Signature Date -------------------------------- ----------------------- ------------- Advisor ---------------------------------- ------------------------- -------------- Co advisor ---------------------------------- -------------------------- ---------------- External Examiner --------------------------------- ---------------------------- ------------ Internal examiner ---------------------------------- ------------------------------ ------------ Chairman i DECLARATION First of all, I declare that this thesis was an outcome of my genuine effort and I submit it to the University of Gondar in partial fulfillment for the requirement of a Degree of Master of Science. The thesis will be placed at the university library to be made accessible to borrowers for reference. I seriously declare that I have not so far submitted this thesis to any other institution anywhere for the reward of any academic degree, diploma, or certificate. Name of the researcher: Abebe Tsega Signature ____________________ Place: University of Gondar Date of submission: _______________________ ii ACKNOWLEDGEMENTS First and foremost, I would like to thank the almighty God for giving me strength to overcome a number of difficulties I faced when I conducted this study. I appreciatively acknowledge my advisor, Dr. Dessie Tibebe, for his unremitting supervision, intelligent guidance and dedication of his time for correction of draft copies. His appreciated advice, encouragement and productive comments that helped me a lot to shape the study in its present form. His approach and regular follow up also played a key role for the successful completion of the experimental work thesis and thesis write-up. Without his help, this research work would have not been accomplished timely. My sincere gratitude also goes to my co-advisor, Mr. Mulugeta Legesse for his excellent technical support during the experimental work, for his valuable comments, guidance on data analysis and important advice towards the completion of this thesis. In addition, I would like to thank laboratory assistants, Mr. Mulat Tiruneh, Mr. Amogne Wondu, Mr. Legesse Terefe and Mrs. Zemenay Alebel for their unreserved support and respectful cooperation during my experimental work. I am very much indebted to my wife, S/r Haregitu Derso for her moral support and encouragement in the course of my study. Finally, my deepest thanks go to the Department of Chemistry, University of Gondar for providing me with the necessary laboratory facilities to conduct my Master’s thesis research. iii TABLE OF CONTENT Titles Page DECLARATION ............................................................................................................................ ii ACKNOWLEDGEMENTS ........................................................................................................... iii LIST OF TABLES ........................................................................................................................ vii LIST OF FIGURES ..................................................................................................................... viii LIST OF ABBREVIATIONS ........................................................................................................ ix ABSTRACT ...................................................................................................................................... x 1. INTRODUCTION ...................................................................................................................... 1 1.1. Background of the Study ...................................................................................................... 1 1.2. Statement of the Problem ..................................................................................................... 3 1.3. Objectives of the Study ........................................................................................................ 4 1.3.1. General Objective .......................................................................................................... 4 1.3.2. Specific Objectives ........................................................................................................ 4 1.4. Significance and Scope of the Study .................................................................................... 4 2. LITERATURE REVIEW ........................................................................................................... 6 2.1. Traditional Medicinal Plants ................................................................................................ 6 2.2. Taxonomy Species and Family of Croton Macrostachyus and Phytolacca Dodecandra ... 7 2.3. The Importance of Medicinal Plants for the Development of Modern Drugs ..................... 8 2.4. Traditional Medicinal Plants in Ethiopia ............................................................................. 9 2.5. Elemental Composition of Traditional Medicinal Plants ................................................... 11 2.6. Essential Macro and Trace Metals ..................................................................................... 11 2.7. Phenolic Contents of Traditional Medicinal Plants............................................................ 14 2.8. Antioxidants ....................................................................................................................... 17 2.9. Medicinal Use of Croton Macrostachyus and Phytolacca Dodecandra............................ 18 iv 2.10. Analytical Techniques for Analysis of Metals in Plant Samples ..................................... 19 2.11. Principle of FAAS ............................................................................................................ 19 2.12. Principle of UV-Vis Spectroscopy ................................................................................... 20 2.13. Principles of Optimization of Digestion Procedures ........................................................ 21 3. MATERIALS AND METHODS .............................................................................................. 22 3.1. Description of the Study Area ............................................................................................ 22 3.2. Apparatus and Reagents ..................................................................................................... 23 3.2.1. Instruments and Apparatus .......................................................................................... 23 3.2.2. Reagents and Chemicals .............................................................................................. 23 3.3. Plant Sampling and Sample Collection Procedures ........................................................... 24 3.4. Sample Pretreatment .......................................................................................................... 25 3.5. Extraction of Medicinal Plants for Total Polyphenolic Content Analysis ......................... 25 3.6. Preparation of Standard Solutions ...................................................................................... 26 3.7. Total Polyphenolic Content Analysis ................................................................................. 27 3.8. Flavonoid Content Analysis ............................................................................................... 27 3.9. Determination of Antioxidant Activity .............................................................................. 28 3.10. Antibacterial activities of the root extracts of medicinal plants ....................................... 28 3.11. Optimization of Digestion Procedure ............................................................................... 29 3.12. Digestion of Croton Macrostachyus and Phytolacca Dodecandra Root Samples .......... 34 3.13. Metal Levels Analysis of Traditional Medicinal plant Root Samples ............................. 34 3.14. FAAS Instrumentation ..................................................................................................... 35 3.15. Instrument Calibration...................................................................................................... 36 3.16. Method Performance and Validation ............................................................................... 37 4. RESULTS AND DISCUSSION ............................................................................................... 40 4.1. Recovery............................................................................................................................. 40 v 4.2. Optimal Conditions for Sample Digestion ........................................................................
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