COMPOSITION OF HAWTHORN (Crataegus spp.) FRUITS AND LEAVES AND EMBLIC LEAFFLOWER (Phyllanthus emblica) FRUITS Pengzhan Liu Department of Biochemistry and Food Chemistry and Functional Foods Forum University of Turku Turku 2012 Supervised by Docent Baoru Yang, Ph.D. Department of Biochemistry and Food Chemistry University of Turku Turku, Finland Professor Heikki Kallio, Ph.D. Department of Biochemistry and Food Chemistry University of Turku Turku, Finland Reviewed by Professor Lothar W. Kroh, Ph.D. Technische Universität Berlin Institut für Lebensmitteltechnologie und Lebensmittelchemie FG Lebensmittelchemie und Analytik Berlin, Germany Docent Kaisu Riihinen, Ph.D. Department of Biosciences University of Eastern Finland Kuopio, Finland Opponent Professor James N. Seiber, Ph.D. Department of Environmental Toxicology University of California, Davis Davis, CA, United States of America ISBN 978-951-29-4981-6 (PRINT) ISBN 978-951-29-4982-3 (PDF) Painosalama Oy – Turku Finland 2012 Content 3 CONTENTS ABSTRACT.....................................................................................................................6 ABBREVIATIONS.........................................................................................................8 LIST OF ORIGINAL PUBLICATIONS......................................................................9 1. INTRODUCTION...................................................................................................10 2. REVIEW OF THE LITERATURE........................................................................12 2.1. Composition of Crataegus spp...........................................................................12 2.1.1. Phenolic compounds in hawthorn of different origins ...........................12 2.1.1.1. Procyanidins.............................................................................12 2.1.1.2. Flavonols and flavonol glycosides............................................14 2.1.1.3. C-glycosyl flavones..................................................................15 2.1.1.4. Other phenolic compounds.......................................................17 2.1.1.5. Contents of phenolic compounds in hawthorn ........................17 2.1.2. Triterpene acids.......................................................................................25 2.1.3. Other compounds....................................................................................25 2.2. Health effects of the phenolic compounds of hawthorn.....................................26 2.2.1. Effects on the heart and vascular system................................................26 2.2.2. Hypolipidaemic effects...........................................................................28 2.2.3. Hypotensive effects.................................................................................29 2.2.4. Antioxidant and radical scavenging activities.........................................30 2.2.5. Anti-inflammatory activities...................................................................33 2.3. Composition of Phyllanthus emblica.................................................................34 2.3.1. Phenolic compounds...............................................................................34 2.3.1.1. Hydrolysable tannins................................................................34 2.3.1.2. Flavonoids and phenolic acids..................................................41 2.3.1.3. Other phenolics.........................................................................43 2.3.2. Norsesquiterpenoids and sesquiterpenoids.............................................45 2.3.3. Vitamin C and other vitamins.................................................................46 2.3.4. Other compounds....................................................................................47 2.4. Health effects of Phyllanthus emblica................................................................47 2.4.1. Antioxidant, radical scavenging and anti-aging activities......................48 2.4.2. Effects on diabetes and diabetic complications.......................................51 2.4.2.1. Effects on type 1 and type 2 diabetes.......................................51 2.4.2.2. Effects on diabetic complications and metabolic syndrome.....52 2.4.3. Effects on cardiovascular health ............................................................55 2.4.4. Anti-inflammatory effects.......................................................................57 4 Content 2.4.5. Cytotoxic effects, protective effects against clastogenicity and anti- cancer activities.......................................................................................59 2.4.5.1. Cytotoxic effects.......................................................................59 2.4.5.2. Protective effects against metal-induced clastogenicity...........60 2.4.5.3. Radioprotective effects.............................................................60 2.4.5.4. Protective effects against chemical-induced carcinogenesis....61 2.4.5.5. Protective effects against the toxicity of anti-cancer medicine.. 61 2.4.6. Effects on the nervous system.................................................................64 2.4.7. Immunomodulating effects.....................................................................65 2.4.8. Anti-microbial and cytotoxic activity.....................................................67 2.4.9. Antidiarrhoeal effects..............................................................................67 2.4.10. Hepatoprotective effects..........................................................................68 2.4.11. Effects on gastric ulceration....................................................................69 2.4.12. Other functions........................................................................................69 3. SUMMARY OF THE PRESENT STUDY.............................................................71 3.1. Objectives of the study.......................................................................................71 3.2. Materials and methods........................................................................................71 3.2.1. Plant samples...........................................................................................71 3.2.2. Analysis of acids, sugars and sugar alcohols in Chinese hawthorn fruits (Study I).........................................................................................72 3.2.2.1. Sample preparation...................................................................72 3.2.2.2. Gas chromatography.................................................................73 3.2.2.3. Qualitative and quantitative analysis........................................73 3.2.3. Analysis of phenolic compounds in hawthorn fruits and leaves (Studies II, III and IV).............................................................................73 3.2.3.1. Sample preparation for qualitative analysis of phenolic compounds in fruits of C. pinnatifida var. major......................73 3.2.3.2. Sample preparation for quantitative analysis of phenolic compounds in fruits of Chinese hawthorn (Study III)..............74 3.2.3.3. Sample preparation for quantitative analysis of fruits and leaves of C. grayana (Study IV)...............................................74 3.2.3.4. HPLC-DAD and HPLC-ESI-MS (Studies II, III and IV).........74 3.2.4. Statistical analysis (Studies I and III)......................................................75 3.2.5. Analysis of phenolic compounds in emblic leafflower fruits..................75 3.2.5.1. Sample preparation...................................................................75 3.2.5.2. HPLC analysis (Study V).........................................................76 3.3. Results and discussion........................................................................................77 3.3.1. Acids, sugars and sugar alcohols in Chinese hawthorn fruits (Study I). 77 3.3.2. Purification and enrichment of phenolic compounds with polyamide column chromatography (Study II).........................................................78 Content 5 3.3.3. Identification of phenolic compounds in Chinese hawthorn fruits (Study II).................................................................................................79 3.3.4. Quantitative analysis of phenolic compounds in hawthorn using HPLC-ESI-MS-SIR (Study III and IV)...................................................80 3.3.4.1. Quantitative analysis of phenolic compounds in Chinese hawthorn fruits (Study III)........................................................81 3.3.4.2. Correlation between the contents of different phenolic compounds in Chinese hawthorn fruits (Study III)..................82 3.3.4.3. Phenolic compounds in fruits and leaves of C. grayana and changes during fruit ripening (Study IV)..................................83 3.3.4.3.1. Fruits.......................................................................................83 3.3.4.3.2. Leaves.....................................................................................84 3.3.5. Composition of phenolic compounds in emblic leafflower fruits (Study V).. 84 3.4. Conclusion..........................................................................................................85 ACKNOWLEDGEMENTS.........................................................................................86 REFERENCES..............................................................................................................88