Polyphenolic and Essential Oil Composition of Mentha and Their Antimicrobial Effect
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KARMEN KAPP Polyphenolic and Essential Oil Composition of Recent Publications in this Series 87/2015 Hongxin Chen Heterobasidion annosum sensu stricto Pathogenesis: Bioinformatic and Functional Study of Cerato-Platanin Family Proteins 88/2015 Kristiina Rajamäki Role of the Inflammasome Pathway in Atherosclerosis 89/2015 Pirkko Kriikku DISSERTATIONES SCHOLAE DOCTORALIS AD SANITATEM INVESTIGANDAM Toxicological Abuse Profile of New Recreational Drugs in Driving-Under-The-Influence and UNIVERSITATIS HELSINKIENSIS 106/2015 Post-Mortem Cases in Finland 90/2015 Eeva Mustonen Red Clover Isoflavonoids in Feed, Plasma and Milk of Ruminants 91/2015 Flavia Restitutti Central, Cardiovascular and Metabolic Effects of Dexmedetomidine Associated with the Selective Peripheral Alpha2-adrenoceptor Antagonist MK-467 in Dogs KARMEN KAPP 92/2015 Katariina Nurmi Prevention of Inflammatory Cellular Responses by Ethanol and Hemin – Interplay Between Inflammasomes and Processes Inhibiting Inflammation Polyphenolic and Essential Oil Composition of 93/2015 Liang He Rare Genetic Variants and Complex Diseases - a Bayesian Approach Mentha and Their Antimicrobial Effect 94/2015 Jani Saksi Stroke-Associated Carotid Atherosclerosis - a Role for FABP4 in Atheroma Vulnerability and Cardiometabolic Risk 95/2015 Otto Hemminki Cancer Immunotherapy with a Gene Modified Serotype 3 Oncolytic Adenovirus 96/2015 Jukka Koskela Individual Trajectories in Asthma and COPD: A Longitudinal Perspective to Obstructive Lung Disease 97/2015 Miira Klemetti Trends In Obstetric and Perinatal Outcomes of Women with Type 1 Diabetes During 1988-2011 – A Finnish Population-Based Observational Study Mentha 98/2015 Ileana B. 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Helsinki 2015 Supervisors Professor Heikki Vuorela, Ph.D. Division of Pharmaceutical Biosciences Faculty of Pharmacy University of Helsinki, Finland Professor Pia Vuorela, Ph.D. Division of Pharmaceutical Biosciences Faculty of Pharmacy University of Helsinki, Finland Professor Tõnu Püssa, Ph.D. Department of Food Hygiene Institute of Veterinary Medicine and Animal Sciences Estonian University of Life Sciences, Estonia Associate Professor Ain Raal, Ph.D. Department of Pharmacy Faculty of Medicine University of Tartu, Estonia Reviewers Professor Anu Hopia, Ph.D. Functional Food Forum Faculty of Medicine University of Turku, Finland Professor Najat A. Saliba, Ph.D. Department of Chemistry Faculty of Arts and Sciences American University of Beirut, Lebanon Opponent Professor Rudolf Bauer, Ph.D. Institute of Pharmaceutical Sciences Department of Pharmacognosy University of Graz, Austria © Karmen Kapp 2015 ISBN 978-951-51-1828-8 (paperpack) ISBN 978-951-51-1829-5 (pdf) ISSN 2342-3161 (print) ISSN 2342-317X (online) Cover illustration F.E. Köhler, Köhler's Medizinal-Pflanzen, Mentha × piperita Hansaprint Printing House Helsinki 2015 TABLE OF CONTENTS ACKNOWLEDGEMENTS LIST OF ORIGINAL PUBLICATIONS CONTRIBUTION OF KARMEN KAPP TO ORIGINAL PUBLICATIONS (I–V) LIST OF ABBREVIATIONS ABSTRACT 1. INTRODUCTION 11 2. REVIEW OF THE LITERATURE 12 2.1. Genus Mentha L. 12 2.1.1. Taxonomy and Mentha plants 12 2.1.2. Polyphenolic composition of Mentha plants 14 2.1.3. Essential oil composition of Mentha plants 15 2.1.4. Variation of the chemical composition of Mentha plants 17 2.1.5. Mentha plants used in food industry 18 2.1.6. Health benefits of Mentha plants 18 2.2. Chlamydia pneumonia 21 2.3. Escherichia coli 22 2.4. Staphylococcus aureus 24 3. AIMS OF THE STUDY 26 4. MATERIALS AND METHODS 27 4.1. Materials 27 4.1.1. Mentha plants 27 4.1.2. Commercial peppermint tea samples 29 4.1.3. Mint flavoured candies and food supplements 31 4.1.4. Reagents and instruments 33 4.2. Methods 34 4.2.1. Extraction of Mentha plants 34 4.2.2. Isolation of Mentha plants essential oils 34 4.2.3. Isolation of mint flavouring hydrodistilled extracts 34 4.2.4 Sample material preparations studied 35 4.2.5. High performance liquid chromatography-mass spectrometry 35 4.2.6. Gas chromatography-mass spectrometry 35 4.2.7. Antichlamydial high-throughput screening 35 4.2.7.1. Cell culture and Chlamydia pneumoniae stock 35 4.2.7.2. Infections 36 4.2.7.3. Infectious progeny assay 36 4.2.7.4. Host cell viability assay 36 4.2.8. Antimicrobial susceptibility testing 37 4.2.8.1. Agar diffusion method 37 4.2.8.2. Broth dilution method 37 4.2.8.2.1. Microbial strains 37 4.2.8.2.2. Broth dilution assay 37 4.2.8.2.3. Determination of minimal bactericidal concentration 38 4.2.9. Statistical analysis 38 3 5. RESULTS AND DISCUSSION 39 5.1. Polyphenolic composition of Mentha plants 39 5.2. Essential oil composition of Mentha plants 41 5.3. Mint flavouring hydrodistilled extract composition 47 5.4. Antibacterial activity 51 5.4.1. Preliminary antibacterial activity tests 51 5.4.2. Antichlamydial activity 51 5.4.2.1. Commercial peppermint tea samples 51 5.4.2.2. Mint flavouring hydrodistilled extracts 51 5.4.3. Activity against Escherichia coli and Staphylococcus aureus 52 5.4.3.1. Water extracts of Mentha plants 52 5.4.3.2. Essential oils of Mentha plants 53 5.4.3.3. Reference substances 54 5.4.3.4. Mint flavouring hydrodistilled extracts 55 6. CONCLUSIONS 56 7. REFERENCES 58 APPENDICES 1-3 ORIGINAL PUBLICATIONS 4 ACKNOWLEDGEMENTS My sincere gratitude goes to my supervisors. I thank Professor Heikki Vuorela for his excellent guidance and encouragement to think and work independently. I am also thankful for his everlasting positive attitude and good humour that supported me throughout the studies. I am also grateful to my supervisor Professor Pia Vuorela for her strong support and her invaluable expertise in the field of pharmaceutical microbiology. Her help in Chlamydia pneumoniae experiments was essential. Special thanks go to my supervisor Professor Tõnu Püssa for devotedly making me acquainted with LC-MS and his great enthusiastic help throughout the years. I would further like to express thanks to supervisor Assoc. Professor Ain Raal who encouraged me to start my doctoral studies and infected me with the high interest towards pharmacognosy; also for introducing the topic of my thesis: Mentha. I would like to thank warmly Dr. Anne Orav from Tallinn University of Technology for her great help and giving the opportunity to learn GC-MS. The co-operation with her was always fluent and I could always rely on her experienced and wise guidance. My sincere thanks go to Docent Päivi Tammela for her excellent guidance and belief in me and my project. Her help was invaluable in the critical moment. I would like to acknowledge Professor Mati Roasto from Estonian University of Life Sciences for his great advice and support. He gave back the faith to my studies. I own special thanks to Docent Leena Hanski for support and encouragement as well as to M.Sc. Elina Hakala and Dr. Terttu Tiirola for their invaluable help with Chlamydia experiments. I thank Intendant Arto Kurtto, Finnish Museum of Natural History, University of Helsinki and Dr. Ülle Reier, Department of Botany, University of Tartu for identifying the damn difficult plants. I am grateful to Professor Jari Yli-Kauhaluoma and Docent Yvonne Holm for evaluating and guiding the progress of my thesis. I would further like to express my gratude to Professor Anu Hopia and Professor Najat A. Saliba for reviewing the manuscript and providing valuable comments and suggestions. I express my sincere thanks to all my colleagues at the Division of Pharmaceutical Biosciences, University of Helsinki for great company and support. Warm thanks to Elina Hakala and Kari Kreander for pleasant atmosphere and chats. I thank also Docent Into Laakso who was always helpful when needed. I would like to thank Krista Virtanen, Heidi Mäkkylä and Tarja Hiltunen for their technical assistance. I would also like to thank the staff from Department of Pharmacy, University of Tartu and Department of Food Hygiene, Estonian University of Life Sciences for their support. TÜ Tamme Pharmacy and Ravana Pharmacy colleagues deserve warm thanks for encouraging me to start the PhD studies. Special thanks to Soile Kuosmanen for her warmhearted and eternal help while settling in to Finland and its culture. Sincere thanks to Anna Klugman for being a brave and inspiring companion during our PhD studies and adventures in Finland. My most sincere and warmest thanks go to my parents, brother and cat Kusti. Thank you for all the support and understanding you have given to finish this study. I am grateful to the Finnish Cultural Foundation (FCF), the Graduate School for Pharmaceutical Research (GSPR) at the University of Helsinki, Chancellor`s Travel Grant and the Society for Medicinal Plant and Natural Product Research (GA) for their financial support.