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EVALUATION of ANTI-ULCER PROPERTIES of METHANOL EXTRACT of Terminalia Superba SEPTEMBER 2014

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EVALUATION of ANTI-ULCER PROPERTIES of METHANOL EXTRACT of Terminalia Superba SEPTEMBER 2014 i EVALUATION OF ANTI-ULCER PROPERTIES of METHANOL EXTRACT of Terminalia superba Engl. & Diels (Combretaceae) STEM BARK A Dissertation Submitted to the Department of Pharmacognosy and Environmental Medicine, Faculty of Pharmaceutical Sciences, University of Nigeria in partial fulfilment of the requirements for the Award of Master of Pharmacy Degree. BY CHUKUMA MICHAEL ONYEGBULAM REG.NUMBER PG/M.PHARM./12/63223. SUPERVISOR: PROFESSOR C.O.EZUGWU SEPTEMBER 2014 ii CERTIFICATION THIS PROJECT REPORT TITLED EVALUATION OF PHYTOCHEMICAL AND ANTI-ULCER PROPERTIES OF Terminalia superba, ENGL. & DIELS (Combretaceae), IS AN ORIGINAL RESEARCH WORK DONE BY CHUKUMA MICHAEL ONYEGBULAM REG.NUMBER PG/M. PHARM./12/63223. IT IS HEREBY CERTIFIED AS MEETING THE REQUIREMENTS FOR THE AWARD OF MASTER OF PHARMACY (M.PHARM) DEGREE OF THE DEPARTMENT OF PHARMACOGNOSY AND ENVIRONMENTAL MEDICINE, FACULTY OF PHARMACEUTICAL SCIENCES, UNIVERSITY OF NIGERIA, NSUKKA. --------------------------------- Prof. C. O. Ezugwu Supervisor/HOD Dept. of Pharmacognosy and Environmental Medicine iii DEDICATION This research is dedicated to the glory of the ONLY LIVING GOD WHO MAKE ALL THINGS BEAUTIFUL IN HIS OWN TIME iv ACKNOWLEDGEMENTS Above all I thank almighty GOD for His protection and provision throughout this course and research work. I thank my family, especially wife Mrs. Harriet Ihuoma Chukuma, for their love, prayers, support and sacrifice towards the success of this project. I cannot thank the following enough for their intellectual input, direction and support- My supervisor Prof. C. O. Ezugwu sir, your patience, availability and readiness to discuss this project have been exceptional, Prof. S. I. Ofoefule, Prof. P.A. Akah, Dr. Mrs. U.E Odo, and Dr. Michele, Mr. Atogba James Mr. Austin N. Okorie, all the staff of the Department of Pharmacognosy and Environmental Medicines. My appreciation of your sacrifice and selfless commitment to this project cannot be sufficiently expressed in words. Without your encouragement and support this project would have been mere wishful thinking. My friends Barr. Jackson Agbai Abba, Mr. Oko Ogbuja, Mr. Rowland Ojukwu, Mr. Nwokocha Obewu, Mr. John Eleanya Onuoha, Miss Deborah Nwanne Ibe, your contributions to the success of this project merit special mention. GOD BLESS YOU ALL. v TABLE OF CONTENTS Title Page ----------------------------------------------------------------------------------- i Certification --------------------------------------------------------------------------------- ii Dedication ----------------------------------------------------------------------------------- iii Acknowledgements ------------------------------------------------------------------------ iv Table of Contents --------------------------------------------------------------------------- v List of Tables-------------------------------------------------------------------------------- vii List of Figures ------------------------------------------------------------------------------ viii Abstract ------------------------------------------------------------------------------------- ix Chapter One ------------------------------------------------------------------------------- 1 1.1 Introduction ------------------------------------------------------------------- 2 1.2 The Plant Terminalia superba ----------------------------------------------- 4 1.2.1 Uses -------------------------------------------------------------------- 9 1.3 Some previous researches on T. superba Engl. & Diels ----------------- 13 1.4 Ulcers -------------------------------------------------------------------------- 17 1.4.1 Pathophysiology of stomach ulcers ---------------------------------- 17 1.4.2 Treatment of ulcers ---------------------------------------------------- 18 1.5 In vivo models used for evaluation of potential anti-gastro duodenal ulcer agents -------------------------------------------------------- 30 1.6 Acute toxicity testing --------------------------------------------------------- 33 Chapter Two: Materials and Methods 2.1 Reagents and Equipments --------------------------------------------------- 34 2.2 Experimental procedure ------------------------------------------------------ 35 2.2.1 Collection, Identification and Preparation--------------------------- 35 vi 2.2.2 Microscopy ------------------------------------------------------------- 35 2.2 3 Extraction --------------------------------------------------------------- 35 2.2.4 Preliminary phytochemical screening of powdered bark ---------- 36 2.2.5 Acute toxicity test of crude extract ----------------------------------- 39 2.2.6 Fractionation ------------------------------------------------------------ 40 2.2.7 Evaluation of antiulcer activity --------------------------------------- 41 Chapter Three: Results 3.1 Macroscopy ------------------------------------------------------------------- 46 3.2 Microscopy -------------------------------------------------------------------- 48 3.3 Phytochemical screening----------------------------------------------------- 54 3.4 Acute toxicity ----------------------------------------------------------------- 58 3.5 Antiulcer activity ------------------------------------------------------------- 59 Chapter Four: Discussion and Conclusion 4.1 Discussion --------------------------------------------------------------------- 64 4.2 Conclusion --------------------------------------------------------------------- 66 References ----------------------------------------------------------------------------------- 67 Appendix ------------------------------------------------------------------------------------ 80 vii LIST OF TABLES Table 1: Some previous researches on T. superba -------------------------------------- 13 Table 2: Review of some anti-ulcer researches involving other plants --------------- 21 Table 3: Some plant derived tannins with antiulcer property -------------------------- 26 Table 4: Some plant derived flavonoids with antiulcer property --------------------- 27 Table 5: Some plant derived saponins with antiulcer property ------------------------ 28 Table 6: Some active constituents isolated from plants -------------------------------- 29 Table 7: Preliminary Phytochemical of Screening of Powdered T. superba Stem Bark ----------------------------------------------------------- 55 Table 8: Phytochemical screening of T superba stem bark crude methanol extract -------------------------------------------------------- 56 Table 9: Phytochemical screening of T superba stem Bark methanol Fraction ---- 57 Table 10: Phytochemical screening of T superba Stem Bark ethyl acetate fraction 58 Table 11: Acute toxicity result ----------------------------------------------------------- 59 Table 12: Antiulcer activity of crude methanol extract of Terminalia superba ----- 61 Table 13: Effect of Methanol Fraction on Aspirin-Induced Ulcer ------------------- 62 Table 14: Comparison of effect of fractions of T. superba stem bark -------------- 63 Table 15: Administration of crude methanol extract ----------------------------------- 80 Table 16: Administration of doses of different fractions ----------------------------- 81 Table 17: Administration of methanol fraction ----------------------------------------- 82 Table 18: Phytochemical screening of T superba stem bark ethyl acetate fraction ---------------------------------------------- 83 viii LIST OF FIGURES Fig 1: Photograph of T. superba --------------------------------------------------------- 6 Fig 2: Internal surface of T. superba stem bark ---------------------------------------- 46 Fig 3: Outside surface of T. superba stem bark ---------------------------------------- 46 Fig 4: Internal surface of T. superba leaf ----------------------------------------------- 47 Fig 5: External surface of T. superba leaf ----------------------------------------------- 47 Fig 6: Branch showing characteristic turfs ---------------------------------------------- 47 Fig 7: Large coloured cork cells---------------------------------------------------------- 48 Fig 8: Fork-shaped fibres ------------------------------------------------------------------ 48 Fig 9: Diamond-shaped Ca oxalate ------------------------------------------------------ 48 Fig 10: Rosette calcium oxalate crystals ----------------------------------------------- 49 Fig 11: Large rosette calcium oxalate crystal ------------------------------------------ 49 Fig 12 Showing sclerieds ----------------------------------------------------------------- 49 Fig 13: Showing fragment of phloem cells --------------------------------------------- 50 Fig 14: Showing fibres with narrow lumen -------------------------------------------- 50 Fig 15: Showing fragment of phloem parenchyma cell -------------------------------- 51 Fig 16: Showing cork cells showing underlying group of fibres ---------------------- 51 Fig 17: Showing large phloem parenchyma cells with calcium oxalate crystals ------------------------------------------------------------------- 52 Fig 18: Showing fragment of phloem cells ---------------------------------------------- 52 Fig 19: Showing phloem parenchyma with biserate medullary ray ------------------- 53 ix ABSTRACT Terminalia superba Engl. & Diels (Combretaceae), is a member of the genus Terminalia that comprises around 100 species distributed in tropical regions of the world. In Africa it is found along the coast of west and central Africa. It has different uses in traditional medicine such as antimalarial, anti-diabetic, anti-fungal, and anti-hypertensive in the areas
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