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Jari Olavi Summanen

A Chemical and Ethnopharmalogical Study on Phyllanthus emblica (Euphorbiaceae)

ACADEMIC DISSERTATION To be presented with the permission of the Faculty of Science of the University of Helsinki, for public criticism in Auditorium XII, on August 4th,1999, at 12 o’clock noon.

University of Helsinki Department of Pharmacy Division of Pharmacognosy

Supervisors: Prof.Raimo Hiltunen, Ph.D. Division of Pharmacognosy Department of Pharmacy University of Helsinki Finland Prof. Heikki Vuorela, Ph.D. Division of Pharmacognosy Department of Pharmacy University of Helsinki Finland Reviewers: Docent Ilkka Ojanperä, Ph.D. Department of Forensic Medicine University of Helsinki Finland Prof. Raimo K. Tuominen, M.D. Division of Pharmacology and Toxicology Department ofPharmacy University of Helsinki Finland Opponent: Prof. Adolf Nahrstedt, Dr. rer. nat. Institut für Pharmazeutische Biologie und Phytochemie der Universität Hittorstraße 56 D-48149 Münster Germany

© Jari Olavi Summanen ISBN 951-45-8677-8 Helsingin yliopiston verkkojulkaisut, 1999


PREFACE ABSTRACT LIST OF ORIGINAL PUBLICATIONS LIST OF ABBREVIATIONS 1. INTRODUCTION 2. AIMS OF THESTUDY 3. REVIEW OF THE LITERATURE 3.1. PHYLLANTHUS 3.1.1. Botanical aspects 3.1.2. Phytochemistry 3.1.3. Ethnopharmacology 3.1.4. Importance of aromatic plant phenols in biological systems 3.1.5. Analysis of phenolic compounds 3.2. SEPARATION METHODS 3.2.1. Planar chromatography 3.2.2. High-performance capillary electrophoresis (CE) 3.2.3. Mobile phase optimization in liquid chromatography 3.3.BIOLOGICAL ASSAYS 3.3.1. Anti-inflammatory activity 3.3.2. Calcium fluxes in cell metabolism 4. EXPERIMENTAL 4.1. MATERIALS 4.2. METHODS 5. RESULTS AND DISCUSSION 5.1. DEVELOPMENT OF ANALYTICAL CHROMATOGRAPHIC METHODS FOR PLANT PHENOLICS 5.1.1. Analysis of phenolic antioxidants by CE (I) 5.1.2. AMD (Automated Multiple Development) optimization (III) 5.1.3. Evaluation of video documentation anddensitometer parameters in the detection of plant phenolics by TLC (III) 5.1.4. Optimization of separation in TLC using desirability functions and mixture designs according to “PRISMA” (IV) 5.2. BIOLOGICAL ACTIVITY 5.2.1. Anti-inflammatory activity of leaf extracts of Phyllanthus emblica L. (V) 5.2.2. Effects of simple aromatic compounds and flavonoids on calcium fluxes in clonal rat pituitary GH4C1cells (VI) 6. CONCLUSIONS 5 7 9 10 12 14 15 15 15 15 18 19 21 21 21 24 25 27 27 28 31 31 32 34 34 34 35 37 40 42 42 43 46





This work was carried out at the Division of Pharmacognosy, Department of Pharmacy, University of Helsinki during the years 1994–1998. I wish to express my gratitude to Professor Raimo Hiltunen, Head of the Pharmacognosy Division andHead of the Department of Pharmacy, for his support during the course of this study and for placing excellent facilities for my work. My sincere gratitude is due to my supervisor, Professor Heikki Vuorela, whose encouraging guidance and stimulating discussions have led my work. Apart from scientific stuff, thanks are also due to openmindedness to discuss anything anytime during all these years. Myspecial thanks go to Professor, M.D. Heikki Vapaatalo, Department of Pharmacology and Toxicology, Institute of Biomedicine, University of Helsinki for introducing me the subject together with Assoc. Prof., Ph.D. M. Zaini Asmawi, Universiti Sains Malaysia, Penang, Malaysia. Special thanks are also due to Docent, M.D. Hannu Kankaanranta, Docent, M.D. Eeva Moilanen and Docent, Ph.D. (Pharm.) PiaVuorela. Without their valuable collaboration my work would never have been started. I want to express my appreciation to Dr. Michael Pasternack for the patch clamp experiments and Prof. Dr. Kid Törnquist, who introduced me to calcium antagonistic test methods. I would also like to thank them for the encouraging discussions and advice throughout the course of the experiments and writing. Mr. Teijo...
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