PHARMACOLOGICAL STUDIES OF SELECTED PLANTS USED TRADITIONALLY IN PAIN AND INFLAMMATION ARSHAD IQBAL DEPARTMENT OF BOTANY ISLAMIA COLLEGE PESHAWAR 2015 PHARMACOLOGICAL STUDIES OF SELECTED PLANTS USED TRADITIONALLY IN PAIN AND INFLAMMATION Thesis submitted in partial fulfillment of the requirements for the award of degree of DOCTOR OF PHILOSOPHY IN BOTANY By ARSHAD IQBAL DEPARTMENT OF BOTANY ISLAMIA COLLEGE PESHAWAR 2015 Dedicated to My Mother TABLE OF CONTENTS TABLE OF CONTENTS ……………………………………………………… ……….. I LIST OF TABLES ……………………………………………………………………..… VI LIST OF FIGURES…………………………………………………………………… IX LIST OF ABBREVIATIONS......................................................................................... XVI ACKNOWLEDGEMENTS…………………………………………………………….. XVIII ABSTRACT ………………………………………………………………………………. 1 1. INTRODUCTION …………………………………………………………………… 03 1.1. Significance and Background …………….................................................................... 03 1.2. The Genus Tamarix ..................................................................................................... 05 1.2.1. Genus Tamarix reported work ....................................................................................06 1.2.1.1. Ethnobotany of Genus Tamarix.............................................................................. 06 1.2.1.2. Phytochemistry of genus Tamarix .......................................................................... 07 1.2.1.3. Pharmacological studies of Genus Tamarix ……………………………………….10 1.3. The Genus Acacia......................................................................................................... 12 1.3.1. Genus Acacia reported work ....................................................................................... 12 1.3.1.1. Ethnobotany of Genus Acacia................................................................................. 11 1.3.1.2. Phytochemistry of Genus Acacia............................................................................ 13 1.3.1.3. Biological studies on Genus Acacia ……………………………………………… 17 1.4. Taxonomical position of Tamarix aphylla. .................................................................. 21 1.4.1. Plant Morphology ..................................................................................................... 21 1.4.2. Distribution ................................................................................................................ 22 1.4.3. Ethnobotanical uses ................................................................................................... 21 1.4.4. Reported isolated compounds of Tamarix aphylla ................................................... 23 1.5. Taxonomical position of Tamarix dioica. ..................................................................... 24 1.5.1. Plant Morphology ....................................................................................................... 24 1.5.2. Distribution ................................................................................................................ 24 1.5.3. Ethnobotanical uses .....................................................................................................25 1.5.4. Reported isolated compounds of Tamarix dioica........................................................ 25 1.6. Taxonomical position of Acacia cyanophylla……………........................................... 26 1.6.1. Plant Morphology ...................................................................................................... 27 I 1.6.2. Distribution ................................................................................................................ 27 1.6.3. Ethnobotanical uses .................................................................................................. 27 1.6.4. Reported isolated compounds of Acacia cyanophylla............................................... 27 1.7. Taxonomical position of Acacia stenophylla………………........................................ 28 1.7.1. Plant Morphology ...................................................................................................... 29 1.7.2. Distribution ............................................................................................................... 29 1.7.3. Ethnobotanical uses ................................................................................................... 29 1.7.4. Reported isolated compounds of Acacia stenophylla................................................. 29 1.8. Natural products ………………………………………………………………………. 30 1.9. Essential oils ……………………………………………………………………………31 1.9.1 Significance of Essential Oils………………………………………………………... 32 1.10. Fixed oils …………………………………………………………………………….. 33 1.11. Pharmacological activities ……………………………………………………………33 1.11.1. Anti-inflammatory activity……………………………………………………..….. 34 1.11.2. Analgesic activity …………………………………………………………..………35 1.11.3. Anti-pyretic activity ………………………………………………………..……… 37 1.11.4. Acute toxicity ……………………………………………………………...………. 37 1.11.5. Anti-microbial activity …………………………………………………..………… 38 1.11.6. Anti-oxidant activity ………………………………………………………………. 38 1.11.7. Enzyme inhibition activity ……………………………………………………...… 39 1.11.8 Aims and objectives………………………………………………………………… 40 2. MATERIALS AND METHODS.................................................................................... 41 2.1. Drugs and Reagents ……….......................................................................................... 41 2.2. General Experimental Conditions ……………............................................................. 42 2.3. Plant Materials ………................................................................................................... 42 2.4. Extraction …….............................................................................................................. 42 2.5. Phytochemical Tests ………......................................................................................... 42 2.5.1. Screening for Different Groups of Compounds ......................................................... 42 2.6. Oils Analysis …….……………………………………………………………………. 45 2.6.1. Extraction …………………………………………………………………………… 45 II 2.6.2. Preparation of Fatty Acid Methyl Esters …………………………………………… 45 2.7. In-Vitro Biological Activities ........................................................................................ 46 2.7.1. Antimicrobial activity …............................................................................................. 46 2.7.2. Acetylcholinesterase inhibition.................................................................................... 50 2.7.3 Lipooxygenase inhibition .............................................................................................50 2.7.4. Antioxidant assay......................................................................................................... 51 2.8. In-Vivo Biological Activities…………...........................................................................51 2.8.1. Experimental animals................................................................................................. 51 2.8.2. Anti-inflammatory activities ………………………………………………………... 51 2.8.2.1 Carageenan Induced Paw Edema Model…………………………………………... 52 2.8.2.2 Xylene-Induced Ear Edema……………………………………………………….. 52 2.8.3 Anti-pyretic activity …………………………………………………………………. 53 2.8.3.1. Pyrexia induction with Brewer‘s Yeast…………………………………………… 53 2.8.4. Analgesic activity.........................................................................................................54 2.8.4.1. Acetic Acid Induced Writhing……………………………………………………. 54 2.8.4.2. Analgesic effect by Hot Plate Method…………………………………………….. 55 2.8.5. Acute toxicity.............................................................................................................. 55 3. RESULTS........................................................................................................................ 56 3.1. Phytochemical Screening (Qualitative)………………….............................................. 56 3.1.1. Phytochemistry of Selected Plants............................................................................... 56 3.2. Oil Analysis.....................................................................................................................57 3.2.1. Oil analysis of Tamarix aphylla................................................................................... 57 3.2.2. Oil analysis of Tamarix dioica …………………........................................................ 58 3.2.3. Oil analysis of Acacia cyanophylla..............................................................................60 3.2.5. Oil analysis of Acacia stenophylla .............................................................................. 62 3.3. In-Vitro Biological Activities……………..................................................................... 64 3.3.1. Antibacterial Activity …………................................................................................. 64 3.3.2. Antifungal activity ...................................................................................................... 81 3.3.3. Acetylcholinesterase inhibitory activities .................................................................. 83 3.3.3.1. Acetylcholinesterase inhibitory activity of Tamarix aphylla………………………83 III 3.3.3.2. Acetylcholinesterase inhibitory activity of Tamarix dioica………………………..83 3.3.3.3. Acetylcholinesterase inhibitory activity of Acacia cyanophylla…………...………84 3.3.3.4.