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By Ugwu Patience Ngozi Pg/M. Pharm/08/49752 Department EFFECTS OF HEAVY METALS ON THE AIR POLLUTION TOLERANCE INDICES ( APTI ) OF FIVE MEDICINAL PLANTS GROWING WITHIN QUARRY SITE IN ISHI-AGU, EBONYI STATE, NIGERIA. BY UGWU PATIENCE NGOZI PG/M. PHARM/08/49752 DEPARTMENT OF PHARMACOGNOSY AND ENVIRONMENTAL MEDICINES, FACULTY OF PHARMACEUTICAL SCIENCES, UNIVERSITY OF NIGERIA, NSUKKA OCTOBER, 2014 TITLE PAGE EFFECT OF HEAVY METALS ON THE AIR POLLUTION TOLERANCE INDICES ( APTI ) OF FIVE MEDICINAL PLANTS GROWING WITHIN QUARRY SITE IN ISHI-AGU, EBONYI STATE, NIGERIA BY UGWU PATIENCE NGOZI PG/M. PHARM/08/49752 BEING REPORT SUMITTED TO THE DEPARTMENT OF PHARMACOGNOSY AND ENVIRONMENTAL MEDICINE, FACULTY OF PHARMACEUTICAL SCIENCES, UNIVERSITY OF NIGERIA, NSUKKA, IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF MASTER OF PHARMACY (M.PHARM) DEGREE. PROF.( MRS) .S.I. INYA-AGHA (SUPERVISOR) DEPARTMENT OF PHARMACOGNOSY AND ENVIRONMENTAL MEDICINES FACULTY OF PHARMACEUTICAL SCIENCES, UNIVERSITY OF NIGERIA, NSUKKA OCTOBER 2014 i CERTIFICATION This project report titled ”Effect of heavy metals on the Air Pollution Tolerance Indices (APTI) of five medicinal plants:Vitex doniana (Lamiceae ), Anthocleista djalonensis (Gentianae), Ricinus communis (Euphorbiacea ), Cnestis ferruginea(Connaraceae) and Manihot esculenta (Euphorbiacea ), growing within quarry site in Ishi-Agu, Ebonyi State Nigeria” is hereby certified as meeting the requirements for the award of Masters of Pharmacy (M.Pharm) degree in the Department of Pharmacognosy and Environmental Medicines, Faculty of Pharmaceutical Sciences,University of Nigeria, Nsukka. ____________________ ____________________ Supervisor Head of Department ____________________ _____________________ Date Date ____________________ External Examiner _________________ Date ii DEDICATION This work is dedicated to my lovely, caring husband (Ugwu Benjamin Sunday) and our children, Ebuka, Ugochukwu and Munachimso. iii ACKNOWLEDGEMENTS My gratitude goes to God Allmighty, whose unfalling love saw me through this programe. I return all the glory to him. My thanks to my supervisor Professor (Mrs) S.I Inya- Agha whose professional advise and support came handy in the period of this duty. I would like to express my warmest gratitude to Dr (Mrs) U.E Odo who graciously attended to my problem when needed. I appreciate the Head of Dept., Professor C.O Ezugwu who provided professional advice and accommodation that facilitated this work. I wish to thank Rev. Dr S.C. Ezea, Mr Otuu Fred. and Pharm M.Ezejiofor, who have always given me their support and assistance. I wish to recognize my classmates, Pharm Edith Atamah , Pharm (Mrs). Okonta, Pharm Okwubie, Pharm Calister Ugwu, Pharm S.A Chime, Pharm Charity Eze for their encouragement. My appreciation goes to the Technical staff of the Department of Pharmacognosy and Environmental Medicines whose numerous technical supports were reassuring in the course of this study. My deepest gratitude goes to my father, Francis Omeje, whose prayerful support gave me strength. My warmest greeting to Franca, Chioma, Kenneth and Chika for their help to see that this work was successful. Our Almighty God will surely reward them. Ugwu patience Ngozi University of Nigeria, Nsuka, 2014. iv ABSTRACT Effects of heavy metals on the Air Pollution Tolerance Indices (APTI) of five medicinal plants growing within quarry site in Ishi-Agu, Ebonyi State, Nigeria were studied. Following laboratory analysis of the leaves of the plants for metal load and four biochemical parameters of APTI computation. The metal load was determined by the Atomic Absorption spectrophotometric method, while the ascorbic acid was determined by the titremetric method, leaf extract pH by the electro-chemical techniques, relative water content by plant turgidity mass and the total chlorophyll, by spectrophotometric method. Foliar photomicrography of the leaf was also studied. Results of Plants from experimental site showed changes in physical and internal structures of the leaves that lead to closure of some stomata pores in test plants and damage of some epicelluler cells. Quantification of phytochemical contents of the test plants showed higher values than that of control which may be due to the multiplication of some organic substances caused by oxidative stress, presence of inorganic subtances like heavy metals and lime stone dust.The result of elemental analysis of both experimental and control plants gave the range of concentration of the metals in parts per million (ppm) as follows: - lead ( 2.75 – 14.13 ± 0.02); Nickle (0.27 - 0.54 ± 0.01) ; Cadmium ( 2.25 - 24.30 ± 0.03); Zinc (0.11- 0.03 ± 0.02); and Arsenic (0.10 - 0.70 ± 0.04). Control –Lead ( 0.00 – 5.52 ± 0.01); Nickle (0.13 – 0.23 ± 0.01); Cadmium (0.00 - 12.00 ± 0.03); Zinc ( 0.06 – 0.10 ± 0.03); and Arsenic (0.02 – 0.20 ± 0.02). There was slight increase of v Cadmium above maximum daily permissible intake in C. ferruginea , A, djalonensis, and R. communis, also there is high content of Arsenic in R.communis which is the cause of oxidative strees in test plants that result in the elicitation of antioxidants from plants and multiplication of ascorbic acid content. The values for the percentage computation of APTI in test plants are V.doniana (11.03), C.ferruginea (7.62), A.djalonesis (9.94), R. communis (11.5) and M. esculenta (8.60). Control pants gave V.doniana (11.86), C.ferruginea (8.21), A.djalonesis (11.86), R. communis (14.39), and M. esculenta (10.21) that result in reduction of Relative water content, Ph value, Total chlorophyll content and Ascorbic acid content.There is relative high content of heavy metals, physiological and visible changes in some sensitive test plants. Owing to information gotten from the result of this research it is not recommended that plants from the quarry sites be used in herbal formulation because of possible metal intoxication and attenuation of their phythopotency. vi TABLE OF CONTENTS Title page: i Certification: ii Dedication: iii Acknowledgements: iv Abstract: v Table of contents: vii List of Tables: x List of Figures: xi CHAPTER ONE: 1 1.1 Introduction 1 1.2 Quarry 2 1.2.1 Effect of quarry on environments 3 1.2.2 Quarry waste 4 1.2.3 Land pollution 5 1.2.4 Noise pollution 5 1.2.5 Damage to biodiversity 6 1.3 Pollution 7 1.3.1 Pollution and vegetation 7 1.3.2 Effect of pollution on leaf anatomy 9 1.4 Air Pollution 11 1.4.1 Air pollution and medicinal plants 13 1.4.2 Types of air pollution, and their sources 14 1.4.3 The threshold which air pollution affects medicinal plants 16 1.4.4 Effect of air pollution on leaf morphology 17 1.4.5 Effect of air pollution on plant sugar content 18 1.4.6 Effects of air pollution on leaf proline 19 1.4.7 General effects of air pollution on medicinal plants 20 1.4.8 Ways of controlling air pollution 23 vii 1.5 Heavy Metals 26 1.5.1 Effect of heavy metals on medicinal plants 27 CHAPTER TWO 29 2.1 Medicinal plants 29 2.2 Pharmacognostic profile of selected medicinal plants 30 2.2.1 Pharmacognostic profile of Cnestis ferruginea 30 2.2.2 Pharmacognostic profile of Ricinus communis 34 2.2.3 Pharmacognostic profile of Vitex doniana 38 2.2.4 Pharmacognostic profile of Anthocliestia djalonensis 41 2.2.5 Pharmacognostic profile of Manihot esculenta 45 2.3 Previous pharmacological invetigation of plants studied 48 2.4 Aim and objective 55 2.5 Significance of study 55 2.6 Definition of terms 56 CHAPTER THREE: MATERIALS AND METHODS 66 3.1 Collection and preparation of plants materials 66 3.2 Chemicals, reagents and equipments 67 3.3 Morphological and microscopic analysis 68 3.4 Qualitative Phytochemical analysis 69 3.5 Quantitative Phytochemical analysis 74 3.6 Determination of Analytical standard 76 3.7 Elemental analysis 80 3.8 Air pollution Tolerance index analysis 84 3.9 Statistical Analysis 86 CHAPTER FOUR: RESULT 87 4.1 Macroscopic /Morphological examination of the leaves of test and control samples 87 4.2 Microscopical examination of the leaves of test and control samples 90 viii 4.3 Summary of the morphological features and microscopic results of leaf from experimental and control sites respectively. 95 4.4 Phytochemical analysis of the leaves of test and control samples 96 4.5 Analytical standard of the plants leaves of the test and control samples 98 4.6 Elemental analysis of the leaves of test and control samples 99 4.7 Relative water contents of the leaves of the leaves of test and control samples 101 4.8 Chlorophyll contents of the leaves of the leaves of test and control samples 102 4.8 pH of the leaf extract of leaves of test and control samples 102 4.9 Ascorbic acid contents of the leaves of test and control samples 103 4.10 Air pollution tolerance indices (APTI) of the leaves of test and control samples 103 CHAPTER FIVE: DISCUSSION AND CONCLUSION 104 5.1 Discussion 104 5.2 Conclusion 116 Recommendation 117 Reference 118 Appendix 127 ix LIST OF TABLES Table1 : Summary of the morphological and microscopic features of leaves of test and control samples 95 Table 2: Qualitative phytochemical analysis of the five medicinal plants 96 Table 3: Quantitative analysis of the five medicinal plants 97 Table 4: Analytical Standard of the five medicinal plants 98 Table 5 : Elemental analysis of the five medicinal plants 100 Table 6 : Relative water contents (RWC) of the five medicinal plants 101 Table 7: Chlorophyll contents of test and control samples 102 Table 8: pH of leaf extract of the five medicinal plants 102 Table 9: Ascorbic acid contents of the five medicinal plants 103 Table 10 : Air pollution tolerance indices (APTI) of the five medicinal
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