Introduction 1 GENERAL CONCEPTS Baby Banana (Musa acuminata AA Simmonds cv. Bocadillo) 4

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1 TABLE OF CONTENTS Introduction 1 1 GENERAL CONCEPTS Baby Banana (Musa acuminata AA Simmonds cv. Bocadillo) Taxonomic Classification of Bananas (Musa acuminata) Plant morphology Pre-harvest Post-harvest Hyperpigmentation phenomenon Chlorophyll degradation High-Speed Countercurrent Chromatography Spiral-Coil Low-Speed Rotary Countercurrent Chromatography (Spiral-Coil LSRCCC) 27 2 RESULTS AND DISCUSSION Phytochemical profile of Baby Banana control peels Phosphatidylcholine (Lecithin) Glycosyldiacylglycerolipids O- -D-Galp (1 6 )- O- -D-Glup (1 3)- 2, 1-diacyl-L-glycerol (GGDG) O- -D-Galp(1-3)-2, 1 diacyl-l-glycerol (MGDG) Compositional analysis of the glycolipid GGDG and MGDG Summary Molecular species of glucocerebroside Sulphoquinovosyl-diacylglycerol (SQDG) XV

2 2.1.5 Phosphatidylethanolamine (PE) Ceramide aminoethylphosphonate (CAEP) Summary Linoleic acid Summary Lutein Violaxanthin-di-laurate and -Carotene isomers Summary Epi-cycloeucalenone and 4-Epi-cyclomusalenone ß-Sitosterol Summary Pheophytin-a Summary Optimization of the solvent system for the separation of chlorophylls and derivatives by means of High-Speed Countercurrent Chromatography in Baby Banana peels, spinach and grass Experimental Design Isolation of chlorophylls and derivatives from spinach by HSCCC Role of dichloromethane and chloroform as stationary phase in the solvent system for the isolating of chlorophylls and xanthophylls from plant extracts by HSCCC Chlorophylls a/b ratio and derivatives in Baby Banana peels and other plants by HSCCC Preparative Spiral-Coil Low Speed Rotary Countercurrent chromatography of Baby Banana peels (Musa acuminata) with hyperpigmentation APCI-HPLC-MS-MS analysis of Spiral-Coil-LSRCCC fractions from Baby Banana peels with hyperpigmentation Identification of polar compounds in Spiral-Coil-LSRCCC 204 XVI

3 Identification of chlorophyll derivatives esterified with sterols in Spiral-Coil-LSRCCC Spiral-Coil-LSRCCC of Baby Banana peels with hyperpigmentation: comparison with HSCCC of Baby Banana control Elucidation of tocopherols and tocotrienols in Fraction 14 of Spiral-Coil-LSRCCC separation from Baby Banana peels with hyperpigmentation extract Elucidation of Triterpene Alcohol Ferulates in Fraction 14 of Spiral-Coil-LSRCCC separation from Baby Banana peels with hyperpigmentation extract Summary SUMMARY AND OUTLOOK MATERIAL AND METHODS Plant Material and Chemicals Plant Material Chemicals and Solvents Equipments and Parameter High-Performance Liquid Chromatography (HPLC) Jasco-System (HPLC-DAD) Knauer-System (preparative) Equipment I Knauer-System (preparative) Equipment II Analytical Columns Preparative Columns Solvent System and Gradients of HPLC DAD Solvent System and Gradients of Preparative HPLC Mass spectrometry APCI-HPLC-MS-MS Column APCI-HPLC-MS-MS 245 XVII

4 Parameter Mode APCI-HPLC-MS-MS Solvent system and gradient of APCI-HPLC-MS High-Speed Countercurrent Chromatography (HSCCC) Solvent System of High-Speed Countercurrent Chromatography (HSCCC) Gas Chromatography GC-MS I GC-MS II GC-MS III Spiral-Coil Low-Speed Rotary Countercurrent Chromatography (Spiral-Coil LSRCCC) Solvent system for Spiral-Coil LSRCCC Nuclear Magnetic Resonance Spectroscopy Methods Extraction Methods First Extraction Method Second Extraction Method Thin Layer Chromatography (TLC) System System Compositional analysis of glycolipids Analysis of fatty acid components by Gas chromatography mass spectrometry (GC-MS) Sugar composition of glycolipids (SCG) Isolation of pheophytin from methanol phase of Baby Banana peels by means of Normal Phase Chromatography 252 Isolation of carotenoids from acetone extract of Baby Banana peels with hyperpigmentation by means of Normal Phase Chromatography XVIII

5 4.3.6 Partial isolation of chlorophyllase and proteins from flavedo of Baby Banana peels SDS/PAGE condition BIBLIOGRAPHY 255 XIX

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