Pharmacognosy Ofiphiona Grantioides (Boiss.) Anderb
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PHARMACOGNOSY OFIPHIONA GRANTIOIDES (BOISS.) ANDERB. AND PLUCHEA ARGUTA BOISS. SUBSP. GLABRA QAISER OF FAMILY ASTERACEAE Ph. D. THESIS BY SHAHIDA NAVEED DEPARTMENT OF BOTANY UNIVERSITY OF PESHAWAR May 2014 PHARMACOGNOSY OFIPHIONA GRANTIOIDES (BOISS.) ANDERB. AND PLUCHEA ARGUTA BOISS. SUBSP. GLABRA QAISER OF FAMILY ASTERACEAE. SHAHIDA NAVEED A THESIS SUBMITTED TO THE DEPARTMENT OF BOTANY, UNIVERSITY OF PESHAWAR, PESHAWAR, PAKISTAN IN PARTIAL FULFILLMENT FOR THE REQUIREMENT OF DEGREE OF DOCTOR OF PHILOSOPHY IN BOTANY DEPARTMENT OF BOTANY UNIVERSITY OF PESHAWAR May 2014 DECLARATION The materials contained within this thesis are my original work and have not been submitted before to this or to any other university. Shahida Naveed CERTIFICATE OF APPROVAL This Dissertation, entitled ―PHARMACOGNOSY OFIPHIONA GRANTIOIDES (BOISS.) ANDERB. AND PLUCHEA ARGUTA BOISS. SUBSP. GLABRA QAISER OF FAMILY ASTERACEAE ‖ submitted by Shahida Naveed is hereby approved and recommended as partial fulfillment for the award of Degree of Doctor of Philosophy in Botany. Professor Dr. Muhammad Ibrar ______________________ Research Supervisor, Department of Botany, University of Peshawar. External _______________________ Examiner Professor Dr. Muhammad Ibrar ________________________ Chairman, Department of Botany, University of Peshawar. DEPARTMENT OF BOTANY UNIVERSITY OF PESHAWAR May 2014 DEDICATED TO MY PARENTS MY HUSBAND AND MY KIDS AHSAN ZUNAIRA MOHSIN LIST OF CONTENTS S .No CONTENTS P. NO Acknowledgement i Abstract iii Publications x CHAPTER-1 INTRODUCTION 1-27 1.1 Pharmacognosy 1 1.2 Medicinal plants 2 1.3 Phytosociology 4 1.4 Ethnobotany 5 1.5 Pharmacognostic study 5 1.5.1 Identification and authentification of plant material 7 1.5.2 Anatomy/Histology 7 1.5.3 Phytochemical analysis 7 1.5.4 Extractive value 8 1.5.5 Fluorescence study 8 1.5.6. Heavy metals analysis 8 1.6. Pharmacology and bioassays 9 1.6.1 Antihelmintic activity 10 1.6.2 Antilice activity 11 1.6.3. Antimicrobial activity 12 1.6.4. Free Radicals and Antioxidants 13 1.6.4. a. Antioxidant activity (DPPH Radical Scavenging activity) 13 1.6.5. Cytotoxicactivity 14 1.6.6. Hepatoprotective activity 16 1.6.7. Insecticidal activity 17 1.6.8. Phytotoxic activity 17 1.7 Family description of research plants 18 1.7.1 Tribe Inuleae 18 1.7.2. Genus Iphiona 18 1.7.3. Iphiona grantioides 18 1.7.4. Taxonomic position Iphiona grantioides (Boiss.). 19 1.7.5. Distribution of Iphiona grantioides (Boiss.) A. Anderb. in Pakistan 20 1.7.6. Ethnomedicinal uses of Iphiona grantioides (Boiss.)A. Anderb. 20 1.8.1 Tribe Plucheeae 22 1.8.2. Genus Pluchea 22 1.8.3. Pluchea arguta (Boiss.). 23 1.8.4. Pluchea arguta .subsp. glabra Qaiser 25 1.8.5. Taxonomic position of Pluchea arguta subsp. glabra. 26 1.8.6. Distribution in Pakistan 27 1.8.7. Ethnomedicinal uses of Pluchea arguta subsp. glabra 27 CHAPTER -2 REVIEW OF LITERATURE 28- 44 2. Review of Literature 28 2.1. Review of literature for Iphiona grantioides 28 2.1.1 Chemical constituents 28 2.1.2 Review of literature for other species of Iphiona 29 2.2. Review of literature for Pluchea arguta subsp. glabra 30 2.3. Review of literature of other species of the two genera i.e., Iphiona and 30 Pluchea (asteraceae) . 2.4. Pharmacognostic studies 36 2.4.1. Ashing 36 2.4.2. Extractive values 37 2.4.3. Fluorescence study 37 2.4.4. Phytochemical analysis 37 2.4.5 Heavy metals analysis 38 2.5 Biological activities 38 2.5.1. Antihelmintic activity 38 2.5.2. Antilice activity 39 2.5.3. Antimicrobial activity 40 2.5.4. Antioxidant activity 41 2.5.5. Cytotoxic activity 42 2.5.6. Hepatoprotective activity 42 2.5.7. Insecticidal activity 43 2.5.8. Phytotoxic activity 44 CHAPTER - 3 MATERIALS AND METHODS 45- 85 3.1 Morphology of the two research plants 45 3.2 Phytosociology 45 3.2.1 Phytosociology of Iphiona grantioidesandPluchea arguta subsp. glabra 45 3.2.2 Community analysis 46 3.2.2.a. Density 46 3.2.2.b. Frequency (%) 47 3.2.2.c. Cover of herbs 48 3.2. 2.d. Importance Value Index 48 3.2.2.e. Determination of Constancy 49 3.3. Soil analysis 49 3.3. 1. Soil sampling 49 3.3.2 Soil texture 49 3.3.3. Soil pH 49 3.3.4. Electrical conductivity 50 3.3.5. Soil organic matter 50 3.3.6. Total Nitrogen 50 3.3.7. Phosphorus 50 3.3.8. Potassium (K) 50 3.4. Ethnobotany 50 3.5. Pharmacognosy 51 3.5.1. Pharmacognosy of Iphiona grantioides and Pluchea arguta subsp. glabra 51 3.5.2. Macroscopical study 51 3.5.3. Microscopic study of leaf, stem and root of Iphiona grantioides and Pluchea 53 arguta subsp. glabra 3.5. 4. Leaf surface study 53 3.5. 4.a. Presence and absence of stomata/ Type of stomata 53 3.5.4.b. Determination of stomatal Number (SN) and Stomatal index (SI) 54 3.5.4.c. Vein islets and vein termination number 55 3.5.4.d. Palisade Cell Ratio 55 3.5.5. Study of physiochemical characteristics of the crud drugs of Iphiona 56 grantioides Pluchea arguta subsp. glabra 3.5.5.a. Ash analysis 56 3.5.5.b. Determination of the moisture 58 3.5.6 Powder drug study of Iphiona grantioides and Pluchea arguta subsp. glabra 59 3.5.7 Phytochemical screening of Iphiona grantioides and Pluchea arguta subsp. 59 glabra. 3.5.8 Florescence study 66 3.5.9. Extractive values in different solvents 67 3.6. Heavy metals analysis 68 3.7. Biological assays 69 3.7.1. Antihelmintic activity. 70 3.7. 2. Anti-lice activity 71 3.7.3. Antibacterial activity 72 3.7.4. Antifungal activity 73 3.7. 5. Antioxidant (DPPH Free radical scavenging ) activity 76 3.7.6. Cytotoxic activity 78 3.7. 7. Hepatoprotective activity 79 3.7.8. Insecticidal activity 82 3.7.9. Phytotoxicity 84 CHAPTER- 4 RESULTS AND DISCUSSION 86- 201 4.1 Morphology of the research plants 86 4.1.1 Morphology of Iphiona grantioides 86 4.1.2 Morphology of Pluchea arguta .subsp. glabra Qaiser 89 4.2 Phytosociology 91 4.2.1 Phytosociology of Iphiona grantioides and Pluchea arguta subsp. glabra 91 4.3 Soil analysis 102 4.3.1 Soil sampling 102 4.3.2 Soil texture 102 4.3.3 Soil pH 102 4.3.4 Electrical conductivity 103 4.3.5 Soil organic matter (OM) 103 4.3.6 Total Nitrogen (N) 103 4.3.7 Available phosphorus (P) 103 4.3.8 Available potassium (K) 103 4.4 Ethnobotany 105 4.4.1 Ethnobotany of Iphiona grantioides 106 4.4.2 Ethnobotany of Pluchea arguta subsp. glabra 107 4.5 Pharmacognosy 107 4.5.1 Pharmacognostic studies of Iphiona grantioides and Pluchea arguta subsp. 107 glabra Qaiser 4.5.2 Macroscopical study of Iphiona grantioides and Pluchea arguta subsp. 108 glabra 4.5.3 Microscopic study of leaf, stem and root of Iphiona grantioides and Pluchea 112 arguta subsp. glabra Qaiser 4.5.3.a. Transverse section (T.S) of Iphiona grantioides stem 112 4.5.3.b Transverse section (T.S) of stem of Pluchea arguta subsp. glabra 115 4..5.3.c. Transverse section (T.S) of Iphiona grantioidesleaf 116 4.5.3.d. Transverse section (T.S.) of Pluchea arguta subsp.glabra. Qaiser leaf 120 4.5.3.e. Transverse section (T.S.) of root of Iphiona grantioides 123 4.5.3.f. Transverse section (T.S.) of root of Pluchea arguta subsp.glabra 124 4.5.4. Leaf surface values of Iphiona grantioides and Pluchea arguta subsp.glabra 126 4.5.4.a. Presence and absence of stomata and types of stomata in Iphiona grantioides 126 and Pluchea arguta subsp.glabra 4.5.4.b Determination of Stomatal number and stomatal index of Iphiona grantioides 126 and Pluchea arguta subsp. glabra 4.5.4.c. Vein islet number and vein termination number of Iphiona grantioides and 127 Pluchea arguta subsp. glabra leaf 4.5.4. d. Palisade ratio of Iphiona grantioides and Pluchea arguta subsp. glabra leaf 127 4.5.5. Physicochemical characteristics of the crude drug of Iphiona grantioides and 130 Pluchea arguta subsp. glabra 4.5.6. a. Powder drug study of different parts of Iphiona grantioides 132 4.5.6.b. Powder drug study of Pluchea arguta subsp. glabra 136 4.5.7 Phytochemical screening 140 4.5.7.a. Qualitative phytochemical screening of leaf, stem, root and flower of Iphiona 140 grantioides 4.5.7.b. Qualitative phytochemical screening of leaves, stem, root of Pluchea arguta 141 subsp. Glabra 4.5.8 Fluorescence study 145 4.5.9 Extractive values 150 4.5.9.a. Extractive values of Iphiona grantioides 150 4.5.9.b Extractive values of Pluchea arguta subsp.glabra 150 4. 6 Heavy metals analysis 151 4.7 Bioassays (Biological activities) of Iphiona grantioides and Pluchea arguta 158 subsp. glabra 4.7.1 Antihelmintic activity 158 4.7.2 Antilice activity 162 4.7.3 Antibacterial activity 165 4.7.4 Antifungal activity 169 4.7.5 Antioxidant activity 176 4.7.6 Cytotoxic activity 178 4.7.7 Hepatoprotective activity 182 4.7.8 Insecticidal activity 190 4.7.9 Phytotoxic activity 197 Conclusion 202 Recommendations 204 References 205 LIST OF FIGURES Fig.No. Title Page No. Fig.1.1 Pediculus humanus capitis (Human Head louse) 12 Fig.1.2. Artemia nauplii (brine shrimps larvae) 15 Fig.1.3. Iphiona grantioides Boiss. 21 Fig.1.3.b. Iphiona grantioides Boiss. 22 Fig.1.4.