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Assesment of Chemical and Morphological Variation of Spanish Marjoram, Spanish Sage and Spike Lavender

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Assesment of Chemical and Morphological Variation of Spanish Marjoram, Spanish Sage and Spike Lavender Departamento de Química Analítica. Facultad de Ciencias TESIS DOCTORAL: ASSESMENT OF CHEMICAL AND MORPHOLOGICAL VARIATION OF SPANISH MARJORAM, SPANISH SAGE AND SPIKE LAVENDER Departamento de Química Analítica. Facultad de Ciencias TESIS DOCTORAL: ASSESMENT OF CHEMICAL AND MORPHOLOGICAL VARIATION OF SPANISH MARJORAM, SPANISH SAGE AND SPIKE LAVENDER Presentada por Inés Méndez Tovar para optar al grado de doctora por la Universidad de Valladolid Co-dirigida por: Dr. Carmen Asensio Sánchez-Manzanera Prof. Dr. Johannes Novak Valladolid, Julio, 2015 “Success is not built on success. It's built on failure. It is built on frustration. Sometimes it is built on catastrophe” A mi familia TABLE OF CONTENTS INDEX OF TABLES....................................................................................1 INDEX OF FIGURES...................................................................................2 ABSTRACT..................................................................................................3 RESUMEN....................................................................................................4 1. GENERAL INTRODUCTION.................................................................5 1.1. AROMATIC AND MEDICINAL PLANTS OVERVIEW 1.2. SECONDARY METABOLITES...................................................9 1.3. LAMIACEAE FAMILY: Lavandula latifolia medik., Salvia lavandulifolia vahl., Thymus mastichina L.........................................12 2. SCOPE OF THE THESIS.......................................................................29 2.1. MOTIVATION AND PURPOSE................................................30 2.2. OBJECTIVES...............................................................................30 2.3. THESIS OUTLINE......................................................................31 3. CHARPTER 1: Volatile fraction differences for Lamiaceae species using different extraction methodologies………………….………...................32 3.1. INTRODUCTION........................................................................32 3.2. METHODS...................................................................................33 3.3. RESULTS AND DISCUSSION...................................................35 3.4. CONCLUSIONS..........................................................................39 4. CHAPTER 2: Variability in essential oil composition of wild populations of Lamiaceae species collected in Spain.................................45 4.1. INTRODUCTION........................................................................46 4.2. METHODS...................................................................................47 4.3. RESULTS.....................................................................................49 4.4. DISCUSSION...............................................................................52 4.5. CONCLUSIONS..........................................................................55 5. CHAPTER 3: By-product of Lavandula latifolia Medik. essential oil distillation as source of natural antioxidants…...........…………………....64 5.1. INTRODUCTION........................................................................65 5.2. METHODS...................................................................................66 5.3. RESULTS.....................................................................................70 5.4. DISCUSSION...............................................................................73 5.5. CONCLUSIONS..........................................................................75 6. CHAPTER 4: Contribution of the main polyphenols of Thymus mastichina to its antioxidant properties L...................................................83 6.1. INTRODUCTION........................................................................84 6.2. METHODS...................................................................................85 6.3. RESULTS AND DISCUSSION...................................................88 6.4. CONCLUSIONS..........................................................................94 7. CHAPTER 5: Polyphenol variability of Salvia lavandulifolia Vahl. Spanish populations………………….......................................................103 7.1. INTRODUCTION......................................................................104 7.2. METHODS.................................................................................104 7.3. RESULTS AND DISCUSSION.................................................106 7.4. CONCLUSIONS........................................................................111 8. CHAPTER 6: Morphological variability of T. mastichina populations................................................................................................121 8.1. INTRODUCTION......................................................................122 8.2. METHODS.................................................................................123 8.3. RESULTS AND DISCUSSION.................................................124 9. GENERAL CONCLUSIONS...............................................................139 10.RESUMEN EN ESPAÑOL..................................................................141 APPENDIX 1: Lamiaceae Photography’s.........................................164 APPENDIX 2: Structural formulas...................................................165 APPENDIX 3: Acknowledgements...................................................167 INDEX OF TABLES Table 1: PhD thesis organizational chart. Table 2: Relative percentage of total peak area of the volatiles obtained with Hydrodistillation (HD), Microdistillation (MD) and Solvent Extraction (EX). Table 3: Main volatile compounds quantification expressed in milligrams of terpenoids per gram of plant material (content > 0.5 mg/g in any of the three methods) and statistical significance of Lavandula latifolia . Table 4: Main volatile compounds quantification expressed in milligrams of terpenoids per gram of plant material (content > 0.5 mg/g in any of the three methods) and statistical significance of Salvia lavandulifolia . Table 5: Main volatile compounds quantification expressed in milligrams of terpenoids per gram of plant material (content > 0.5 mg/g in any of the three methods) and statistical significance of Thymus mastichina . Table 6: Geographical coordinates of the studied populations. Table 7: Minimum, maximum and average of the compounds found in the essential oil of 11 populations of Thymus mastichina L. collected in 2009, 2010 and 2011 expressed in peak area percentage (%). Statistical significance for year and population variables. Table 8: Average, maximum and minimum amount of the compounds found in the essential oil of 12 populations of Salvia lavandulifolia from 2009, 2010 and 2011 expressed in peak area percentage (%).Statistical significance for year and population variables. Table 9: Average, maximum and minimum amount of the compounds found in the essential oil of 12 populations of Lavandula latifolia collected in 2009, 2010 and 2011 expressed in peak area percentage (%).Statistical significance for year and population variables. Table 10: Quality ranges from the International Standard Organization for Thymus mastichina, Salvia lavandulifolia and Lavandula latifolia . Table 11: Geographical coordinates of collected populations of Lavandula latifolia . Table 12: Total phenol content, EC 50 of scavenging activity, and EC 50 of ferric reduction antioxidant power, of the plant and the by-product of the 12 population of Lavandula latifolia . Table 13: Percentages of the sum of squares obtained in the analysis of variance by type of plant material using population, year and year by population interaction as sources of variation. Table 14: Coefficients of Pearson correlation among total phenols, free radical scavenging activity and ferric reducing antioxidant power. Table 15: Phenolic compounds identified in by-product of 12 Lavandula latifolia populations (results expressed in mg/100 g of dry extract). Table 16. Stepwise regression analysis taking scavenging activity (DPPH) and Ferric Reduction Antioxidant Power (FRAP) as dependent variables and the polyphenol content of apigenin, luteolin and rosmarinic acid as independent variables. Table 17. Data of the origins of the analyzed Thymus mastichina populations. Table 18. Pearson correlation coefficients between the antioxidative test and the identified polyphenols. Table 19: Stepwise regression analysis taking scavenging activity (DPPH), ferric reduction antioxidant power (FRAP) and total phenols (TP) as dependent variables and the polyphenol contents as independent variables. Table 20 : Data of the origins of the Salvia lavandulifolia populations. Table 21: Amounts of detected compounds in the 7 populations of Salvia lavandulifolia . Table 22 : Maximum, minimum, average and deviation for the antioxidant activity colorimetric methodologies. Table 23. Significative correlations between methods, original coordinates and peaks. Table 24: Data of the origins of the studied populations of Thymus mastichina . Table 25: Qualitative and quantitative traits studied in Thymus mastichina . Table 26: Mean, range, coefficients of variation (CVs) and main components for 20 morphological traits in 14 populations of Thymus mastichina collected in 2011. Table 27: Pearson correlation coefficients among the 20 studied traits of Thymus mastichina . Table 28: Percentages of the sum of squares obtained in the analysis of variance using population, year and year x population
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