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Studies on the Standardization of Rose Water and Assessment of Its Uses/ Role in Herbal Medicine

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Studies on the Standardization of Rose Water and Assessment of Its Uses/ Role in Herbal Medicine STUDIES ON THE STANDARDIZATION OF ROSE WATER AND ASSESSMENT OF ITS USES/ ROLE IN HERBAL MEDICINE By SAFIA ABIDI B.Pharm., M.Pharm. DEPARTMENT OF PHARMACOGNOSY FACULTY OF PHARMACY AND PHARMACEUTICAL SCIENCES UNIVERSITY OF KARACHI KARACHI-75270, PAKISTAN 2018 STUDIES ON THE STANDARDIZATION OF ROSE WATER AND ASSESSMENT OF ITS USES/ ROLE IN HERBAL MEDICINE By SAFIA ABIDI B.Pharm., M.Pharm. Thesis submitted for the partial fulfillment of the degree of Doctor of Philosophy DEPARTMENT OF PHARMACOGNOSY FACULTY OF PHARMACY AND PHARMACEUTICAL SCIENCES UNIVERSITY OF KARACHI KARACHI-75270, PAKISTAN 2018 ii CERTIFICATE I have gone through the thesis titled “STUDIES ON THE STANDARDIZATION OF ROSE WATER AND ASSESSMENT OF ITS USES / ROLE IN HERBAL MEDICINE” submitted by Miss Safia Abidi for the award of Ph.D. degree and certify that to the best of my knowledge it contains no plagiarized material. Signature & Seal of Supervisor Prof. Dr. Iqbal Azhar Department of Pharmacognosy Faculty of pharmacy and pharmaceutical Sciences University of Karachi Date : iii BOARD OF ADVANCED STUDIES & RESEARCH UNIVERSITY OF KARACHI DECLARATION I, SAFIA ABIDI d/o SYED TAJAMMUL HUSAIN hereby declare that no part of the work presented by me has been plagiarized from anywhere. Proper references are cited wherever necessary. I understand that the university reserves the right to cancel the degree if any of the above declaration is proved false before or even after the award of degree. Signature of Candidate:…………………………….. Name of the Candidate: SAFIA ABIDI Title of the thesis: “Studies on the Standardization of Rose Water and Assessment of Its Uses / Role in Herbal Medicine” Degree: Ph.D. Department: Pharmacognosy Date:……………………………. iv IN THE NAME OF ALLAH THE MOST BENEFICENT THE MOST MERCIFUL v DEDICATION Dedicated to my parents for their continous support To my husband for be my strength and To my priceless loving sons Aarib and Muhammad vi TABLE OF CONTENTS Pg. No. List of Tables xi List of Figures xv List of Abbreviations xviii Acknowledgements xx Abstract xxii Urdu Translation xxv PART-Ia 1. INTRODUCTION 2 1.1 Introduction PART-Ib LITERATURE REVIEW 7 1.2 Introduction 8 1.3 Morphology 8 1.4 History 9 1.5 Varieties of Rosa damascena 10 1.6 Phytoconstituents of Rosa damascena 10 1.7 Products obtained from Rosa damascena Petals 12 1.8 Traditional Uses of Rosa damascena 12 1.9 Current Uses of Rosa damascena 13 1.10 Toxicity 14 PART-II 2. MATERIAL AND METHODS 15 2.1 List of Chemical and Reagents Used 16 2.2 List of Apparatus/ Equipment Used 18 2.3 List of Microorganisms 20 2.4 List of Animals 21 2.5 List of Solution Used 21 vii TABLE OF CONTENTS Pg. No. 2.6. Collection and Identification of Rose Species (Flower) 22 2.7. Preparation of Rose water 23 2.8. Evaluation of Physicochemical Parameters 24 2.8.1 Color 24 2.8.2 pH 24 2.8.3 Detection of Saponin 24 2.8.3.1 Foam Test 24 2.8.4 Detection of Tannins 24 2.8.4.1 Lead Acetate Test 24 2.8.4.2 Nitric Acid Test 25 2.8.5 Detection of Triterpenoids 25 2.8.5.1 Liebermann Burchard Test 25 2.8.6 Detection of Fixed Oil 25 2.8.6.1 Spot Test 25 2.8.7 Detection of Flavanoids 25 2.8.7.1 Lead Acetate Test 25 2.8.7.2 Sulphuric Acid Test 25 2.9 Determination of Volatile Constituents by HS-GC-MS 26 2.10 Detection of Functional Groups by FT-IR 26 2.11 Acute toxicity Studies 26 2.11.1 Experimental Animals 26 2.11.2 Experimental Design 27 2.12 Brine Shrimp(Artemia Salina) Lethality Bioassay 27 2.12.1 Shrimp’s Eggs Viability Test 27 2.12.2 Brine Shrimp Lethality Assay 27 2.12.3 Data Analysis 28 2.13 In-Vitro Anti-Oxidant Activity 28 2.13.1 Preparation of Stock Solution 28 viii TABLE OF CONTENTS Pg. No. 2.13.2 Determination of Reducing Power 28 2.13.3 Statistical Analysis 29 2.14 In-Vitro Anti-Inflammatory Activity 29 2.14.1 Preparation of Stock Solution 29 2.14.2 Inhibition of Albumin Denaturation Method 30 2.14.2.1 Control Solution (50ml) 30 2.14.2.2 Standard solution (50ml) 30 2.14.2.3 Test solution (50ml) 30 2.15 Determination of In-vitro Sun Protecting Factor (SPF) 31 2.16 Anti-Microbial Activity 32 2.16.1 Collection of Isolates 32 2.16.2 Standard Anti-Microbial Agents 33 2.16.3 Anti-Microbial Assay 34 2.17 Microbial Contamination 34 2.17.1 Sample preparation 34 2.17.2 Bacterial contamination 34 2.18 Clinical Studies 35 2.18.1 Clinical Protocol 35 2.19 Glow Measurement 35 2.19.1 Study Protocol 35 2.19.2 Instrumentation 36 2.19.3 Method 36 2.19.4 Statistical Analysis 36 2.20 Determination of Skin Hydration and Oil Content 36 2.20.1 Study Design 36 2.20.2 Statistical analysis 37 2.21 Cream Formulation 37 2.21.1 Placebo Cream and Its Preparation 37 ix TABLE OF CONTENTS Pg. No. 2.21.2 Cream Formulation and Its Preparation 38 2.22. In-vitro Anti-Inflammatory Activity of Rose Water Based Cream 41 Formulation 2.22.1 Preparation of Stock Solution 41 2.22.2 Inhibition of albumin denaturation method 41 2.23 In -vitro Anti-oxidant Activity of Rose Water Based Cream Formulation 42 2.23.1 Preparation of stock solution 42 2.23.2 Determination of reducing power 42 2.24 Antitussive Activity of Rose Water Based Cough Syrup Formulation 43 2.24.1 Preparation of cough syrup formulations 43 2.25 Screening for Antitussive activity of Cough Syrup 45 2.25.1 Animals Used 46 2.25.2 Method 46 PART-III 3. RESULTS 48 3.1 Evaluation of Physicochemical Parameters 48 3.2 Determination of Volatile Constituents by HS-GC-MS 48 3.3 Detection of Functional Groups by FT-IR 49 3.4 Acute Toxicity Study 49 3.5 Brine Shrimp(Artemia Salina) Lethality Bioassay 49 3.6 In-vitro Anti-Oxidant Assay by Ferric Reducing Power 49 3.7 In-vitro Anti-Inflammatory Activity 50 3.8 Determination of In-vitro Sun Protecting Factor (SPF) 50 3.9 Anti-Microbial Activity 51 3.10 Microbial Contamination 51 3.11. Clinical Studies 52 3.12 Glow Measurement of Skin 52 3.13 Measurement of Skin Hydration and Oil Content 52 x TABLE OF CONTENTS Pg. No. 3.14 In-vitro Anti-Inflammatory activity of Rose Water Based Cream 53 Formulation 3.15 In-vitro Anti-Oxidant Activity of Rose Water Based Cream 53 Formulation 3.16 Antitussive Activity of Rose Water Based Cough Syrup Formulation 54 PART-IV 4. DISCUSSION 56 4.1 Evaluation of Physicochemical Parameters 56 4.2 Determination of Volatile Constituents by HS-GC-MS 57 4.3 Detection of Functional Groups by FT-IR 58 4.4 Acute Toxicity Study 58 4.5 Brine Shrimp(Artemia Salina) Lethality Bioassay 49 4.6 In-vitro Anti-Oxidant Assay by Ferric Reducing Power 60 4.7 In-vitro Anti-Inflammatory Activity 62 4.7.1 Chronic Inflammation 62 4.7.2 Recent advancement on inflammation 63 4.8 Determination of In-vitro Sun Protecting Factor (SPF) 65 4.9 Anti-Microbial Activity 67 4.10 Microbial Contamination 67 4.11 Clinical Studies 68 4.12 Glow Measurement of Skin 68 4.13 Measurement of Skin Hydration and Oil Content 69 4.14 In-vitro Anti-Oxidant Activity of Rose Water Based Cream 70 Formulation 4.15 In-vitro Anti-Inflammatory Activity of Rose Water Based Cream 71 Formulation 4.16 Antitussive Activity of Rose Water Based Cough Syrup Formulation 72 xi TABLE OF CONTENTS Pg. No. PART-V CONCLUSION AND SUGGESTIONS 75 REFERENCES 181 PUBLICATION 200 xii LIST OF TABLES Table No. Title of Tables Pg. No. Part I (a) Table 1.1 General testing parameters for the standardization of herbal 5 medicine Part II (b) Table 1.2.1 Chemical composition of rose essential oil obtained from 10 different parts of the world Table 1.2.2 Traditional uses of different parts of rose flower 12 Table 1.2.3 Pharmacological effects of different types of extracts of Rose 13 flower Part-II Table 2.1 Normalized product function used in the calculation of SPF 32 Table 2.2 Composition of placebo cream 39 Table 2.3 Composition of cream formulation 40 Table 2.4 Composition of cough syrups formulation C1 and C2 44 Table 2.5 Composition of cough syrups formulation C3 and C4 45 Table-3 pH Trend of Rose Water Samples 77 Table-4 Phytochemical Screening of Rose Water Samples 78 Table-5 Volatile Components Present in Rose Water Samples 79 Table-6 FT-IR Absorption Band Assignments of Rose Water Samples 80 Table-7 Acute Toxicity Study of Rose Water at 15, 20,25ml Adult Dose 81 Table-8 Brine Shrimp (Artemia salina) Lethality Bioassay of Sample #1 82 Table-9 Brine Shrimp (Artemia salina) Lethality Bioassay of Sample #2 82 Table-10 Brine Shrimp (Artemia Salina) Lethality Bioassay of Sample #3 83 Table-11 Brine Shrimp (Artemia salina) Lethality Bioassay of Sample #4 83 Table-12 Brine Shrimp (Artemia salina) Lethality Bioassay of Sample #5 84 Table-13 Brine Shrimp (Artemia salina) Lethality Bioassay of Sample #6 84 Table-14 Brine Shrimp (Artemia salina) Lethality Bioassay of Sample #7 85 Table-15 Brine Shrimp (Artemia salina) Lethality Bioassay of Sample #8 85 xiii Table No.
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