UNIVERSITAT POLITÈCNICA DE VALÈNCIA Departament de Tecnologia d'Aliments DOCTORAL THESIS OBTENTION AND CHARACTERIZATION OF ROSEMARY AND ASH TREE SEED EXTRACTS AND STUDY OF THEIR PREVENTIVE EFFECTS ON METABOLIC DISORDERS Presented by: Sixto Alvin IBARRA BOBER Directed by: Dr. Amparo CHIRALT BOIX Dr. Marc ROLLER València, 2011 i ii DOCTORAL THESIS OBTENTION AND CHARACTERIZATION OF ROSEMARY AND ASH TREE SEED EXTRACTS AND STUDY OF THEIR PREVENTIVE EFFECTS ON METABOLIC DISORDERS Presented by: Sixto Alvin IBARRA BOBER Ingeniero Alimentario UNIVERSIDAD FEDERICO VILLARREAL (LIMA – PERU) Magister en Ciencia de los Alimentos UNIVERSIDAD DE CHILE Presented at: UNIVERSITAT POLITÈCNICA DE VALÈNCIA To obtain the degree: “DOCTOR PER LA UNIVERSITAT POLITÈCNICA DE VALÈNCIA” Directed by: Dr. Amparo CHIRALT BOIX Profesora catedrática de la Universidad Politécnica de Valencia Dr. Marc ROLLER Scientific Director of NATUREX València, 2011 iii iv Acknowledgments To v vi Dedicatory To my family, friends and colleagues vii viii INDEX Page ABBREVIATIONS - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 1 ABSTRACT - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 3 RESUMEN - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 5 RESUM - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 7 INTRODUCTION - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 9 OBJECTIVES - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 17 CHAPTER I: IMPORTANCE OF EXTRACT STANDARDIZATION AND IN VITRO/EX VIVO ASSAY SELECTION FOR THE EVALUATION OF ANTIOXIDANT ACTIVITY OF BOTANICALS: A CASE STUDY ON THREE Rosmarinus officinalis L. EXTRACTS- - - - - - - - - - - - - - - - - - - - - 19 CHAPTER II: CARNOSIC ACID-RICH ROSEMARY (Rosmarinus officinalis L.) LEAF EXTRACT LIMITS WEIGHT GAINS AND IMPROVES CHOLESTEROL LEVELS AND GLYCAEMIA IN MICE ON A HIGH-FAT DIET - - - - - - - - - - - - - - - - - - - - -- - - - - - - - - - - - - - - - - - - 41 CHAPTER III: IRIDOIDS FROM Fraxinus excelsior L. WITH ADIPOCYTE DIFFERENTIATION-INHIBITORY AND PPARa ACTIVATION 61 ACTIVITY - - - - - - - - - - - - - - - - - - - -- - - - - - - - - - - - - - - - - - - - - - - - - CHAPTER IV: Fraxinus excelsior L. SEED EXTRACT FRAXIPURE™ LIMITS WEIGHT GAINS AND HYPERGLYCEMIA IN HIGH-FAT DIET- 79 INDUCED OBESE MICE - - - - - - - - - - - - - - - - - - -- - - - - - - - - - - - - - - - CHAPTER V: ACUTE EFFECTS OF Fraxinus excelsior L. SEED EXTRACT ON POSTPRANDIAL GLYCEMIA AND INSULIN 97 SECRETION ON HEALTHY VOLUNTEERS - - - - - - - - - - - - - - - - - - -- - DISCUSSION - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 119 CONCLUSIONS- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 129 ANNEX - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 131 ix x