Technische Universität München Lehrstuhl für Chemisch-Technische Analyse und Chemische Lebensmitteltechnologie Technological separation and analysis of flavanones from different plants and their microbiological activity Iwona Małgorzata Proczek Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grades eines Doktor-Ingenieurs genehmigten Dissertation. Vorsitzender: Univ.-Prof. Dr. K.-H. Engel Prüfer der Dissertation: 1. Univ.-Prof. Dr. Dr. h.c. H. Parlar 2. Univ.-Prof. Dr. Dr. h.c. A. K. Bledzki (Universität Kassel) Die Dissertation wurde am 26.01.2011 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 20.06.2011 angenommen. First and foremost, I would like to express my gratitude to Prof. Dr. Dr. Harun Parlar, not only for the interesting subject material and making my dissertation possible, but also for his professional guidance, helpful advice, financial assistance and continuous support during all aspects of this thesis. I wish to express my sincere appreciation to Prof. Dr. Karl-Heinz Engel from the Department of General Food Technology kindly accepting the position as chairman in my examination, and to Prof. Dr. Andrzej Bledzki, from the University in Kassel, for being my second thesis reviewer. I am also grateful to Dr. Frank Otto for answering many of my questions on various applications, for the fruitful discussions, for his encouragement, patience, a big aid and supervision all throughout the thesis also from a distance. You should know that without your help it would be very difficult for me to write this thesis. I want to emphasize a special thanks to Albrecht Friess for the incredible atmosphere during these 3 years, and especially for proofreading my thesis. Thank you for providing me with a lot of helpful, advice and for the continuous willingness to help. I would like to express my thanks to Dr. Vogel, and especially his successor Dr. Mathias Hutzler from the Institute for Brewing and Food Quality, for kindly lending me a laboratory, for the active collaboration, and the patient understanding. My special thanks also go to Margit Gramma from the Institute for Brewing and Food Quality for helping me with the analyses, her smile which always brought a nice atmosphere, and her continuous readiness to help. I also owe many thanks to Dr. Ludwig M. Niessen from the Department of Technical Microbiology for allowing me to use the SunRise Tecan and also Patrick Preissler for his kindly help by analyses. I am also indebted to Dr. Martin Haslbeck, from the Department of Biotechnology in Garching, for making the circular dichroism analyses possible, and for all the help and advice. I would like to thank Dr Thomas Letzel for allowing me to use the mass spectrometer, and Silvia Grosse for her support during the analyses and the pleasant atmosphere at the office. I want to emphasize a special thanks to Dr. Mehmed Colhan for his help and kind words in difficult moments. Many thanks also to Daniela Schossig and Julia Meineke, from the RAPS Forschung Zentrum, for wonderful cooperation. Thanks to Dr. Johanna Graβmann, Romy Scheerle and Dr. Florian Weiland for sharing the office and nice environment. I also owe many thanks to Jutta Pierschalik, Claudia Steinmetz and Irene Goros for providing me with the various documents, gracious aid and wonderful atmosphere. I would also like to express many thanks to Dr. Eng. Lorenz Gabel for his help, especially in the initial phase of my work. Your friendship and friendly environment, created by the multitude of interesting conversations, were unforgettable. I want to emphasize with all my heart my gratitude to Tanja Weber for every moment during these 3 years! I cannot express in words how much you did for me, not only by helping me to solve a huge number of different problems during the creation of this work, but also by your smile, friendship and kind heart that was always open to listen and help in any circumstances! I would like to thank Yvonne Hanrahan for reading and correcting parts of this thesis. I also owe a great deal of gratitude to Krystyna and Zdzisław Stańczyk, and their daughter Moni for your support and for all the time I spent at your home. This thesis would not have been possible without you, my dear Friends! I am thankful to all my friends in Munich, who have made me feel at home here, and to my friends abroad, for the trips, visits and e-mails, reminding me that friendship remains despite any distance. To write about your help and support would be overwhelmingly lengthy, so I decided to only mention your names. I hope we will make more moments to remember! I would like to thank Agatka Miłosz, Angelica Liguori, Angels Via Estern, Ania Kalita, Ania Górka-Babik, Berenika Zaraska, Beril Caylak, Chiara Alfano, Cony Wendler Vidal, Cris Burqueño, Elunia Romej, Gilda Fulco, Elena Serrano Bertos, Kasia Mikła, Martita Lopez, Liz Costa, Madzia Różańska, Madzia Kuty, Marylka Neubauer, Miriam Olejnik, Monia Dworecka, Misia Kołodziej, Nuria Riera, Paola Azzarino, Paula Gil, Renijka Załęcka, Sandrita Berdala, Vale Angeletti, Antonio Sala, Balazs Matuz, Carlos Gomez Bartolomé, Chen Teng, Dejan Pangercic, Domenico Lorusso, Edu Aguilar Moreno and his wonderful Family, Franio Lazaro Blasco, Giuliano Garrammone, Fr. Jan Kruczyński, Javi Bueno García, Javi Mulero, Jordi Rivera, José Gardiazabal, José Garzon, Fr. Józef Zborzil, Juanito Cabezas García, Juli Cocera Cañas, Maciuś Tusz, Marco Ristic, Marek Babik, Paweł Telega, Pau Goldstein, Rafa Rodriguez, Raul Tejedor, Ricardo Minguez, Romain Hermenier, Silvio Pasquali, Thomas Wendler Vidal, Tomaso de Cola and Yves Strittmatter. I want to emphasize special thanks to Benjamin Gaczkowski for his support, understanding and encouragement especially in the worst moments of writing of this thesis! I want to thank you for your help and time, simply being there for me! Most of all I want to thank all my family, especially my phenomenal brothers Mariusz and Piotr for being my teachers from the beginning, for their support, help at any time with all things I needed also from a distance! I would like to thank their wives Marta and Madzia, as well for being with us and making the family and the time spent together richer and beautiful!! At the end I want to thank my parents, the most important people! If I would like to write here all things for that I owe to thank them, I wouldn't have enough space, but if I wrote just a few situations, it wouldn't express everything what they did! I write then just one sentence: Thank you from all my heart for your Love, Love which was a source of everything and brought me everything!!! All wisdom comes from the LORD and with him it remains forever. The sand of the seashore, the drops of rain, the days of eternity: who can number these? Heaven's height, earth's breadth, the depths of the abyss: who can explore these? Before all things else wisdom was created; and prudent understanding, from eternity. To whom has wisdom's root been revealed? Who knows her subtleties? There is but one, wise and truly awe-inspiring, seated upon his throne: It is the LORD; he created her, has seen her and taken note of her. He has poured her forth upon all his works, upon every living thing according to his bounty; he has lavished her upon his friends…. Sir 1, 1-10 Dedicated to my wonderful Family Contents 1. Introduction ................................................................................................... 1 1.1. The topic of the thesis ................................................................................................ 3 2. Theoretical Background ............................................................................... 5 2.1. Flavonoids ................................................................................................................... 5 2.1.1 Structure and nomenclature of flavonoids .......................................................... 6 2.1.2. Occurrence of flavonoids .................................................................................... 7 2.1.3. Medicinal properties of flavonoids ..................................................................... 8 2.1.4. Human therapeutic significance of flavonoids ................................................... 9 2.1.5. Absorption and metabolism of flavonoids ........................................................ 10 2.1.6. Antioxidant properties of flavonoids ................................................................ 11 2.1.7. Tumors protective activity of flavonoids .......................................................... 12 2.1.8. Flavonoids as immune modulators ................................................................... 13 2.1.9. Antiviral activity of flavonoids ......................................................................... 14 2.1.10. Antimicrobial activity of flavonoids ................................................................. 15 2.1.11. Toxicity of flavonoids....................................................................................... 16 2.2. Flavanone ................................................................................................................. 17 2.2.1. Naringenin ........................................................................................................ 18 2.2.2. Isosakuranetin ..................................................................................................