Printed Edition of the Special Issue Published in Marine Drugs Advances in Marine Chitin and Chitosan Edited by Hitoshi Sashiwa and David Harding www.mdpi.com/journal/marinedrugs Hitoshi Sashiwa and David Harding (Eds.) Advances in Marine Chitin and Chitosan This book is a reprint of the special issue that appeared in the online open access journal Marine Drugs (ISSN 1660-3397) in 2014 (available at: http://www.mdpi.com/journal/marinedrugs/special_issues/advance_chitin). Guest Editors David Harding Massey University New Zealand Hitoshi Sashiwa Kaneka C RUSRUDWLRQ Japan Editorial Office MDPI AG Klybeckstrasse 64 Basel, Switzerland Publisher Shu-Kun Lin Managing Editor Tina Yan 1. Edition 2015 MDPI • Basel • Beijing • Wuhan ,6%1 3') ISBN 978-3-03842-1 (Hbk) © 2015 by the authors; licensee MDPI, Basel, Switzerland. All articles in this volume are Open Access distributed under the Creative Commons Attribution .0 license (http://creativecommons.org/licenses/by/ .0/), which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. However, the dissemination and distribution of p h ysical copies of this book as a whole is restricted to MDPI, Basel, Switzerland. III Table of Contents List of Contributors ............................................................................................................ VII About the Guest Editors...................................................................................................... XII Preface .............................................................................................................................. XIII &KDSWHU, &KHPLVWU\ Priyanka Sahariah, Vivek S. Gaware, Ramona Lieder, Sigríður Jónsdóttir, Martha Á. Hjálmarsdóttir, Olafur E. Sigurjonsson and Már Másson The Effect of Substituent, Degree of Acetylation and Positioning of the Cationic Charge on the Antibacterial Activity of Quaternary Chitosan Derivatives Reprinted from: Mar. Drugs 2014 , 12 (8), 4635-4658 http://www.mdpi.com/1660-3397/12/8/4635 .......................................................................... 3 Syang-Peng Rwei, Yu-Ming Chen, Wen-Yan Lin and Whe-Yi Chiang Synthesis and Rheological Characterization of Water-Soluble Glycidyltrimethylammonium-Chitosan Reprinted from: Mar. Drugs 2014 , 12 (11), 5547-5562 http://www.mdpi.com/1660-3397/12/11/5547....................................................................... 27 Inmaculada Aranaz, María C. Gutiérrez, María Luisa Ferrer and Francisco del Monte Preparation of Chitosan Nanocompositeswith a Macroporous Structure by Unidirectional Freezing and Subsequent Freeze-Drying Reprinted from: Mar. Drugs 2014 , 12 (11), 5619-5642 http://www.mdpi.com/1660-3397/12/11/5619....................................................................... 43 Sudipta Chatterjee, Fabien Salaün and Christine Campagne The Influence of 1-Butanol and Trisodium Citrate Ion on Morphology and Chemical Properties of Chitosan-Based Microcapsules during Rigidification by Alkali Treatment Reprinted from: Mar. Drugs 2014 , 12 (12), 5801-5816 http://www.mdpi.com/1660-3397/12/12/5801....................................................................... 69 IV George Z. Kyzas and Dimitrios N. Bikiaris Recent Modifications of Chitosan for Adsorption Applications: A Critical and Systematic Review Reprinted from: Mar. Drugs 2015 , 13 (1), 312-337 http://www.mdpi.com/1660-3397/13/1/312 .......................................................................... 85 Nidal Daraghmeh, Babur Z. Chowdhry, Stephen A. Leharne, Mahmoud M. H. Al Omari and Adnan A. Badwan Co-Processed Chitin-Mannitol as a New Excipient for Oro-Dispersible Tablets Reprinted from: Mar. Drugs 2015 , 13 (4), 1739-1764 http://www.mdpi.com/1660-3397/13/4/1739 ...................................................................... 111 Shubin Yang, Dadong Shao, Xiangke Wang, Guangshun Hou, Masaaki Nagatsu, Xiaoli Tan, Xuemei Ren and Jitao Yu Design of Chitosan-Grafted Carbon Nanotubes: Evaluation of How the – OH Functional Group Affects Cs + Adsorption Reprinted from: Mar. Drugs 2015 , 13 (5), 3116-3131 http://www.mdpi.com/1660-3397/13/5/3116 ...................................................................... 137 &KDSWHU,, 0HGLFLQDO%LRORJ\ Riccardo A. A. Muzzarelli, Mohamad El Mehtedi and Monica Mattioli-Belmonte Emerging Biomedical Applications of Nano-Chitins and Nano-Chitosans Obtained via Advanced Eco-Friendly Technologies from Marine Resources Reprinted from: Mar. Drugs 2014 , 12 (11), 5468-5502 http://www.mdpi.com/1660-3397/12/11/5468..................................................................... 155 Shao-Jung Wu, Trong-Ming Don, Cheng-Wei Lin and Fwu-Long Mi Delivery of Berberine Using Chitosan/Fucoidan-Taurine Conjugate Nanoparticles for Treatment of Defective Intestinal Epithelial Tight Junction Barrier Reprinted from: Mar. Drugs 2014 , 12(11), 5677-5697 http://www.mdpi.com/1660-3397/12/11/5677..................................................................... 191 Jing-Yi Hou, Li-Na Gao, Fan-Yun Meng and Yuan-Lu Cui Mucoadhesive Microparticles for Gastroretentive Delivery: Preparation, Biodistribution and Targeting Evaluation Reprinted from: Mar. Drugs 2014 , 12 (12), 5764-5787 http://www.mdpi.com/1660-3397/12/12/5764..................................................................... 213 V Jin-Kyu Rhee, Ok Kyu Park, Aeju Lee, Dae Hyeok Yang and Kyeongsoon Park Glycol Chitosan-Based Fluorescent Theranostic Nanoagents for Cancer Therapy Reprinted from: Mar. Drugs 2014 , 12 (12), 6038-6057 http://www.mdpi.com/1660-3397/12/12/6038..................................................................... 237 Qing-Xi Wu, Dong-Qiang Lin and Shan-Jing Yao Design of Chitosan and Its Water Soluble Derivatives-Based Drug Carriers with Polyelectrolyte Complexes Reprinted from: Mar. Drugs 2014 , 12 (12), 6236-6253 http://www.mdpi.com/1660-3397/12/12/6236..................................................................... 257 Moacir Fernandes Queiroz, Karoline Rachel Teodosio Melo, Diego Araujo Sabry, Guilherme Lanzi Sassaki and Hugo Alexandre Oliveira Rocha Does the Use of Chitosan Contribute to Oxalate Kidney Stone Formation? Reprinted from: Mar. Drugs 2015 , 13 (1), 141-158 http://www.mdpi.com/1660-3397/13/1/141 ........................................................................ 276 Toril Andersen, Stefan Bleher, Gøril Eide Flaten, Ingunn Tho, Sofia Mattsson and Nataša Škal ko-Basnet Chitosan in Mucoadhesive Drug Delivery: Focus on Local Vaginal Therapy Reprinted from: Mar. Drugs 2015 , 13 (1), 222-236 http://www.mdpi.com/1660-3397/13/1/222 ........................................................................ 294 Adnan A. Badwan, Iyad Rashid, Mahmoud M. H. Al Omari and Fouad H. Darras Chitin and Chitosan as Direct Compression Excipients in Pharmaceutical Applications Reprinted from: Mar. Drugs 2015 , 13 (3), 1519-1547 http://www.mdpi.com/1660-3397/13/3/1519 ...................................................................... 309 Nidal A. Qinna, Qutuba G. Karwi, Nawzat Al-Jbour, Mayyas A. Al-Remawi, Tawfiq M. Alhussainy, Khaldoun A. Al-So'ud, Mahmoud M. H. Al Omari and Adnan A. Badwan Influence of Molecular Weight and Degree of Deacetylation of Low Molecular Weight Chitosan on the Bioactivity of Oral Insulin Preparations Reprinted from: Mar. Drugs 2015 , 13 (4), 1710-1725 http://www.mdpi.com/1660-3397/13/4/1710 ...................................................................... 340 VI Zakieh I. Al-Kurdi, Babur Z. Chowdhry, Stephen A. Leharne, Mahmoud M. H. Al Omari and Adnan A. Badwan Low Molecular Weight Chitosan – Insulin Polyelectrolyte Complex: Characterization and Stability Studies Reprinted from: Mar. Drugs 2015 , 13 (4), 1765-1784 http://www.mdpi.com/1660-3397/13/4/1765 ...................................................................... 356 &KDSWHU,,, 2WKHUV Woo-Jin Jung and Ro-Dong Park Bioproduction of Chitooligosaccharides: Present and Perspectives Reprinted from: Mar. Drugs 2014 , 12 (11), 5328-5356 http://www.mdpi.com/1660-3397/12/11/5328..................................................................... 379 Tzu-Wen Liang, Chih-Ting Huang, Nguyen Anh Dzung and San-Lang Wang Squid Pen Chitin Chitooligomers as Food Colorants Absorbers Reprinted from: Mar. Drugs 2015 , 13 (1), 681-696 http://www.mdpi.com/1660-3397/13/1/681 ........................................................................ 409 Islem Younes and Marguerite Rinaudo Chitin and Chitosan Preparation from Marine Sources. Structure, Properties and Applications Reprinted from: Mar. Drugs 2015 , 13 (3), 1133-1174 http://www.mdpi.com/1660-3397/13/3/1133 ...................................................................... 425 VII List of Contributors Inmaculada Aranaz: Instituto de Ciencia de Materiales de Madrid (ICMM), Consejo Superior de Investigaciones Científicas (CSIC), (Materials Science Institute of Madrid, Spanish National Research Counsil), Cantoblanco 28049, Madrid, Spain. Toril Andersen: Drug Transport and Delivery Research Group, Department of Pharmacy, Faculty of Health Sciences, University of Tromsø The Arctic University of Norway, Tromsø 9037, Norway. Nawzat Al-Jbour: The Jordanian Pharmaceutical Manufacturing Company (PLC), Research and Innovation Centre, P.O. Box 94, Naor 11710, Jordan. Mayyas A. Al-Remawi: College of Pharmacy, Taif University, Taif 5700, Saudi Arabia. Tawfiq M. Alhussainy: Department of Pharmacology and Biomedical Sciences, Faculty of Pharmacy and Medical Sciences, University of Petra, P.O. Box 96134, Amman 11196, Jordan. Khaldoun A. Al-So'ud: Department of Chemistry, Faculty of Science, Al al-Bayt University, P.O. Box 130040, Mafraq 25113, Jordan. Mahmoud M. H. Al Omari: The Jordanian Pharmaceutical Manufacturing Company (PLC), Research and Innovation Center (RIC), Suwagh Subsidiary for Drug Delivery Systems, P.O. Box 94, Naor 11710, Jordan. Zakieh I. Al-Kurdi: The Jordanian Pharmaceutical Manufacturing Company (PLC), Suwagh Subsidiary for Drug Delivery Systems, P.O. Box 94, Naor 11710, Jordan; Faculty of Engineering & Science, University of Greenwich, Medway Campus, Chatham Maritime, Kent ME44TB, UK. Adnan A. Badwan: The Jordanian Pharmaceutical Manufacturing Company (PLC), Suwagh Subsidiary for Drug Delivery Systems, P.O. Box 94, Naor 11710, Jordan; Research and Innovation Center (RIC), The Jordanian Pharmaceutical Manufacturing Co., P.O. Box 94, Naor 11710, Jordan. Stefan Bleher: Drug Transport and Delivery Research Group, Department of Pharmacy, Faculty of Health Sciences, University of Tromsø The Arctic University of Norway, Tromsø 9037, Norway. Dimitrios N. Bikiaris: Laboratory of Polymer Chemistry and Technology, Division of Chemical Technology, Department of Chemistry, Aristotle University of Thessaloniki, GR- 541 24 Thessaloniki, Greece. Yu-Ming Chen: Institute of Organic and Polymeric Materials, National Taipei University of Technology, 1, Sec. 3, Zhongxiao E. Rd., Taipei 10648, Taiwan. Whe-Yi Chiang: Institute of Organic and Polymeric Materials, National Taipei University of Technology, 1, Sec. 3, Zhongxiao E. Rd., Taipei 10648, Taiwan. Yuan-Lu Cui: Research Center of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. VIII Sudipta Chatterjee: University of Lille Nord de France, F-59000 Lille, France; ENSAIT/GEMTEX, F-59100 Roubaix, France. Christine Campagne: University of Lille Nord de France, F-59000 Lille, France; ENSAIT/GEMTEX, F-59100 Roubaix, France. Babur Z. Chowdhry: Faculty of Engineering & Science, University of Greenwich, Medway Campus, Chatham Maritime, Kent ME44TB, UK. Francisco Del Monte: Instituto de Ciencia de Materiales de Madrid (ICMM), Consejo Superior de Investigaciones Científicas (CSIC), (Materials Science Institute of Madrid, Spanish National Research Counsil), Cantoblanco 28049, Madrid, Spain. Trong-Ming Don: Department of Chemical and Materials Engineering, Tamkang University, New Taipei City 251, Taiwan. Nguyen Anh Dzung: Institute of Biotechnology & Environment, Tay Nguyen University, Buon Ma Thuot 63000, Vietnam. Fouad H. Darras: Research and Innovation Center (RIC), The Jordanian Pharmaceutical Manufacturing Co., P.O. Box 94, Naor 11710, Jordan. Nidal Daraghmeh: The Jordanian Pharmaceutical Manufacturing Co., PO Box 94, Naor 11710, Jordan; Faculty of Engineering & Science, University of Greenwich, Medway Campus, Chatham Maritime Kent ME44TB, UK. Gøril Eide Flaten: Drug Transport and Delivery Research Group, Department of Pharmacy, Faculty of Health Sciences, University of Tromsø The Arctic University of Norway, Tromsø 9037, Norway. Moacir Fernandes Queiroz: Department of Biochemistry, Biosciences Centre, Federal University of Rio Grande do Norte, Salgado Filho avenue 3000, Natal, RN 59078-970, Brazil. María Luisa Ferrer: Instituto de Ciencia de Materiales de Madrid (ICMM), Consejo Superior de Investigaciones Científicas (CSIC), (Materials Science Institute of Madrid, Spanish National Research Counsil), Cantoblanco 28049, Madrid, Spain. Vivek S. Gaware: Faculty of Pharmaceutical Sciences, School of Health Sciences, University of Iceland, Hofsvallagata 53, IS-107 Reykjavík, Iceland; PCI Biotech AS, Strandveien 55, N- 1366 Lysaker, Norway. María C. Gutiérrez: Instituto de Ciencia de Materiales de Madrid (ICMM), Consejo Superior de Investigaciones Científicas (CSIC), (Materials Science Institute of Madrid, Spanish National Research Counsil), Cantoblanco 28049, Madrid, Spain. Li-Na Gao: Research Center of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. Martha Á. Hjálmarsdóttir: Department of _Biomedical Science, Faculty of_ Medicine, University of _Iceland, Stapi, Hringbraut 31, 101 Reykjavík, Iceland. Jing-Yi Hou: State Key Laboratory of Earth Surface Processes and Resources Ecology, Beijing Normal University, Haidian District, Beijing 100088, China; State Key Laboratory of Dao-di Herbs, China Academy of Chinese Medical Sciences, Beijing 100700, China. Guangshun Hou: Institute of Resources & Environment, Henan Polytechnic University, Jiaozuo 454000, China. IX Chih-Ting Huang: Department of Chemistry, Tamkang University, New Taipei City 25137, Taiwan. Woo-Jin Jung: Division of Applied Bioscience & Biotechnology, Institute of Environment- Friendly Agriculture (IEFA), College of Agricultural and Life Sciences, Chonnam National University, Gwangju 500-757, Korea. Sigríður Jónsdóttir: Department of Chemistry, Science Institute, University of Iceland, Dunhagi 3, IS-107 Reykjavik, Iceland. George Z. Kyzas: Laboratory of Polymer Chemistry and Technology, Division of Chemical Technology, Department of Chemistry, Aristotle University of Thessaloniki, GR-541 24 Thessaloniki, Greece. Qutuba G. Karwi: Department of Pharmacology and Biomedical Sciences, Faculty of Pharmacy and Medical Sciences, University of Petra, P.O. Box 96134, Amman 11196, Jordan. Ramona Lieder: The REModeL Lab, The Blood Bank, Landspitali University Hospital, Snorrabraut 60, 105 Reykjavik, Iceland. Stephen A. Leharne: Faculty of Engineering & Science, University of Greenwich, Medway Campus, Chatham Maritime Kent ME44TB, UK. Wen-Yan Lin: Institute of Organic and Polymeric Materials, National Taipei University of Technology, 1, Sec. 3, Zhongxiao E. Rd., Taipei 10648, Taiwan. Cheng-Wei Lin: Department of Biochemistry and Molecular Cell Biology, School of medicine, Taipei Medical University, Taipei City 110, Taiwan; Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei City 110, Taiwan. Dong-Qiang Lin: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China. Tzu-Wen Liang: Life Science Development Center, Tamkang University, No. 151, Yingchuan Rd., Tamsui, New Taipei City 25137, Taiwan; Department of Chemistry, Tamkang University, New Taipei City 25137, Taiwan. Aeju Lee: Biomedical Research Center, Korea Institute of Science and Technology, Seoul 136-791, Korea. Riccardo A. A. Muzzarelli: Faculty of Medicine, Department of Clinical & Molecular Sciences, Polytechnic University of Marche, IT-60100 Ancona, Italy. Mohamad El Mehtedi: Faculty of Engineering, Department of Industrial Engineering & Mathematical Sciences, Polytechnic University of Marche, IT-60100 Ancona, Italy. Monica Mattioli-Belmonte: Faculty of Medicine, Department of Clinical & Molecular Sciences, Polytechnic University of Marche, IT-60100 Ancona, Italy. Fwu-Long Mi: Department of Biochemistry and Molecular Cell Biology, School of Medicine, Taipei Medical University, Taipei City 110, Taiwan; Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei City 110, Taiwan. X Fan-Yun Meng: State Key Laboratory of Earth Surface Processes and Resources Ecology, Beijing Normal University, Haidian District, Beijing 100088, China; State Key Laboratory of Dao-di Herbs, China Academy of Chinese Medical Sciences, Beijing 100700, China. Karoline Rachel Teodosio Melo: Department of Biochemistry, Biosciences Centre, Federal University of Rio Grande do Norte, Salgado Filho avenue 3000, Natal, RN 59078-970, Brazil. Sofia Mattsson: Department of Pharmacology and Clinical Neuroscience, Division of Clinical Pharmacology, Umeå University, Umeå SE-90187, Sweden. Már Másson: Faculty of Pharmaceutical Sciences, School of Health Sciences, University of Iceland, Hofsvallagata 53, IS-107 Reykjavík, Iceland. Masaaki Nagatsu: Graduate School of Science and Technology, Shizuoka University, 3-5-1, Johoka-ku, Hamamatsu 432-8561, Japan. Ro-Dong Park: Division of Applied Bioscience & Biotechnology, Institute of Environment- Friendly Agriculture (IEFA), College of Agricultural and Life Sciences, Chonnam National University, Gwangju 500-757, Korea Ok Kyu Park: Division of Bio-imaging, Chuncheon Center, Korea Basic Science Institute, Gangwon-do 200-701, Korea. Kyeongsoon Park: Division of Bio-imaging, Chuncheon Center, Korea Basic Science Institute, Gangwon-do 200-701, Korea. Nidal A. Qinna: Department of Pharmacology and Biomedical Sciences, Faculty of Pharmacy and Medical Sciences, University of Petra, P.O. Box 96134, Amman 11196, Jordan. Xuemei Ren: School of Environment and Chemical Engineering, North China Electric Power University, Beijing 102206, China. Syang-Peng Rwei: Institute of Organic and Polymeric Materials, National Taipei University of Technology, 1, Sec. 3, Zhongxiao E. Rd., Taipei 10648, Taiwan. Jin-Kyu Rhee: Western Seoul Center, Korea Basic Science Institute, Seoul 120-140, Korea. Hugo Alexandre Oliveira Rocha: Department of Biochemistry, Biosciences Centre, Federal University of Rio Grande do Norte, Salgado Filho avenue 3000, Natal, RN 59078-970, Brazil. Marguerite Rinaudo: Biomaterials Applications, 6 rue Lesdiguières, Grenoble 38000, France. Iyad Rashid: Research and Innovation Center (RIC), The Jordanian Pharmaceutical Manufacturing Co., P.O. Box 94, Naor 11710, Jordan. Priyanka Sahariah: Faculty of Pharmaceutical Sciences, School of Health Sciences, University of Iceland, Hofsvallagata 53, IS-107 Reykjavík, Iceland. Olafur E. Sigurjonsson: The REModeL Lab, The Blood Bank, Landspitali University Hospital, Snorrabraut 60, 105 Reykjavik, Iceland;Institute of Biomedical and Neural Engineering, Reykjavik University, Menntavegur 1, 101 Reykjavik, Iceland. Fabien Salaün: University of Lille Nord de France, F-59000 Lille, France; ENSAIT/GEMTEX, F-59100 Roubaix, France. XI Diego Araujo Sabry: Department of Biochemistry, Biological Sciences Centre, Federal University of Parana, Coronel Francisco H. dos Santos avenue S/N, Curitiba, PR CP 19031, Brazil; Department of Biochemistry, Molecular Biology, Federal University of São Paulo, Três de Maio street 100, São Paulo, SP 04044-020, Brazil. Guilherme Lanzi Sassaki: Department of Biochemistry, Biological Sciences Centre, Federal University of Parana, Coronel Francisco H. dos Santos avenue S/N, Curitiba, PR CP 19031, Brazil. Nataša Š kalko-Basnet: Drug Transport and Delivery Research Group, Department of Pharmacy, Faculty of Health Sciences, University of Tromsø The Arctic University of Norway, Tromsø 9037, Norway. Dadong Shao: School of Environment and Chemical Engineering, North China Electric Power University, Beijing 102206, China. Ingunn Tho: PharmaLuxLab Research Group, School of Pharmacy, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway. Xiaoli Tan: School of Environment and Chemical Engineering, North China Electric Power University, Beijing 102206, China. Shao-Jung Wu: Department of Chemical Engineering, Ming Chi University of Technology, New Taipei City 243, Taiwan. Qing-Xi Wu: Integrated Biotechnology Laboratory, School of Life Science, Anhui University, Hefei 230601, China; Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China. San-Lang Wang: Life Science Development Center, Tamkang University, No. 151, Yingchuan Rd., Tamsui, New Taipei City 25137, Taiwan; Department of Chemistry, Tamkang University, New Taipei City 25137, Taiwan. Xiangke Wang: School of Environment and Chemical Engineering, North China Electric Power University, Beijing 102206, China; School for Radiological and Interdisciplinary Sciences, Soochow University, Suzhou 215123, China; Collaborative Innovation Center of Radiation Medicine of Jiangsu Higher Education Institutions, Suzhou 215123, China. Dae Hyeok Yang: Institute of Cell & Tissue Engineering, College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea. Shubin Yang: School of Environment and Chemical Engineering, North China Electric Power University, Beijing 102206, China; Graduate School of Science and Technology, Shizuoka University, 3-5-1, Johoka-ku, Hamamatsu 432-8561, Japan. Shan-Jing Yao: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China. Islem Younes: Laboratory of Enzyme Engineering and Microbiology, University of Sfax, National School of Engineering, PO Box 1173-3038, Sfax, Tunisia. Jitao Yu: Institute of Resources & Environment, Henan Polytechnic University, Jiaozuo 454000, China. XII About the Guest Editors David Harding was born in London, England in 1944. After completing a B.Sc. (Honors) at the University of Canterbury, Christchurch, New Zealand in 1967, he took up a research position with the American drug company, Eli Lilly at their Erl Wood research center in Windlesham, Surrey, England. In 1969, he began a Ph.D. program at the University of Western Ontario, London, Ontario, Canada. On the completion of his PhD study in 1973, he returned to New Zealand to take up a position at Massey University, Palmerston North. His current title is Professor of Separation Science. He is a synthetic organic chemist with over 100 publications and patents. He has associated interests in analytical chemistry. His association with cellulose in the 1990s led to a commercial product (MEP-cellulose), now sold by the Pall Company for the purification of antibodies using a technique he developed called HCIC (hydrophobic charge induction chromatography). The HCIC program led a special Massey University research award and still returns royalties to the University. His polysaccharide interests in natural polysaccharides have focused on chitosan from the late 1990s. Currently sodium alginate and hyaluronic acid also feature in his research programs. He is a member of the New Zealand Association of Scientists and a Fellow of the New Zealand Institute of Chemistry. Hitoshi Sashiwa was born in Osaka, Japan, in 1963. He received his Ph.D. degree from Hokkaido University (Japan) under the supervision of Professor S. Tokura in 1991. He worked at Tottori University (Japan) as Assistant Associated Professor from 1988 to 2000. He worked with Professor R. Roy at the University of Ottawa (Canada) for two years (1998 í 2000). He worked at AIST Kansai (Japan) as a postdoctoral scholar during 2000 í 2004. He has been affiliated with Kaneka Co. Ltd. (Japan) since April 2004. His research interests include chemical modification of chitin and chitosan and their biomedical applications. He is a member of The Society of Polymer Science, Japan, and the Japanese Society for Chitin and Chitosan. He is the sole author of 70 publications and co-author of 30 publications. XIII Preface Recently, biomass-based polymers from renewable resources have received increasing focus owing to the depletion of petroleum resources. Natural polysaccharides such as cellulose, hemicellulose, and starch are among the candidates from natural resources for biomass polysaccharide products including bioplastics. Although several kinds of neutral or anionic polysaccharides such as chitin, alginic acid, hyaluronic acid, heparin, and chondroitin sulfate exist in nature, natural cationic polysaccharides are quite limited. Chitin is second only to cellulose as the most natural abundant polysaccharide in the world. Chitosan, the product from the N -deacetylatation of chitin, appears to be the only natural cationic polysaccharide. Therefore, chitin and chitosan due to their unique properties are expected to continue to offer a vast number of possible applications for not only chemical or industrial use, but also biomedicine. The research history on chitins, one of the most major and abundant natural polysaccharides on earth, started around 1970. Since the 1980s, chitin and chitosan research (including D-glucosamine, N -acetyl-D-glucosamine, and their oligomers) has progressed significantly over several stages in both fundamental research and industrial fields. With the opening of this book, we planned to produce a strong, very exciting issue that will encompass breakthroughs in highly valuable, scientific, and industrial research in this field. This book covers recent trends in all aspects of basic and applied scientific research on chitin, chitosan and their derivatives. Hitoshi Sashiwa and David Harding Guest Editors Chapter I: Chemistry 3 The Effect of Substituent, Degree of Acetylation and Positioning of the Cationic Charge on the Antibacterial Activity of Quaternary Chitosan Derivatives Priyanka Sahariah, Vivek S. Gaware, Ramona Lieder, Sigríður Jónsdóttir, Martha Á. Hjálmarsdóttir, Olafur E. Sigurjonsson and Már Másson Abstract: A series of water-soluble cationic chitosan derivatives were prepared by chemoselective functionalization at the amino group of five different parent chitosans having varying degrees of acetylation and molecular weight. The quaternary moieties were introduced at different alkyl spacer lengths from the polymer backbone (C-0, C-2 and C-6) with the aid of 3,6-di- O - tert -butyldimethylsilyl protection of the chitosan backbone, thus allowing full (100%) substitution of the free amino groups. All of the derivatives were characterized using 1 H-NMR, 1 H- 1 H COSY and FT-IR spectroscopy, while molecular weight was determined by GPC. Antibacterial activity was investigated against Gram positive S. aureus and Gram negative E. coli . The relationship between structure and activity/toxicity was defined, considering the effect of the cationic group’s structure and its distance from the polymer backbone, as well as the degree of acetylation within a molecular weight range of 7–23 kDa for the final compounds. The N , N , N -trimethyl chitosan with 100% quaternization showed the highest antibacterial activity with moderate cytotoxicity, while increasing the spacer length reduced the activity. Trimethylammoniumyl quaternary ammonium moieties contributed more to activity than 1-pyridiniumyl moieties. In general, no trend in the antibacterial activity of the compounds with increasing molecular weight or degree of acetylation up to 34% was observed. Reprinted from Mar. Drugs . Cite as: Sahariah, P.; Gaware, V.S.; Lieder, R.; Jónsdóttir, S.; Hjálmarsdóttir, M.Á.; Sigurjonsson, O.E.; Másson, M. The Effect of Substituent, Degree of Acetylation and Positioning of the Cationic Charge on the Antibacterial Activity of Quaternary Chitosan Derivatives. Mar. Drugs 2014 , 12 , 4635-4658. 1. Introduction Chitin is a structural polysaccharide that forms the basic constituent of the outer skeleton of insects and crustaceans, including shrimps and crabs. Chitin can be partially or fully deacetylated using strong alkali to give chitosan. Chitosan is therefore a heteropolysaccharide comprised of 2-amino-2-deoxy- D -glucopyranose (glucosamine) and N -acetyl glucosamine units linked through (1 ĺ 4)- ȕ -glycosidic bonds. A number of applications have been found for chitosan in the fields of pharmaceutics [1], biomedicine [2], cosmetics [3] and the food industry [4], due to its unique combination of various properties, like bioactivity, biocompatibility, biodegradability and lack of toxicity [5,6]. Amongst its various properties, the antimicrobial efficacy and applications of chitosan against bacteria have been the focus of many investigations. Chitosan has limited solubility in aqueous media above pH 6. It shows antibacterial properties only in acidic media. This activity is not observed at high pH, due to the absence of the positively charged amino groups and also due to low 4 solubility in aqueous media [7–9]. Chitosan derivatives, in which permanent positive charges were introduced onto the polymer backbone, have been synthesized, which led, in general, to good aqueous solubility and also contributed to significant antibacterial activity at neutral pH [10]. Previously, such derivatives have been prepared by quaternizing the amino group of native chitosan [11,12] or by introducing the quaternized group in one step through an acylation or alkylation reaction [13,14]. This leads to products that are heterogeneous with respect to the degree of substitution (DS) on the amino group and often partially O -modified [15]. Regioselective triphenylmethyl (trityl) protection of the primary (C-6) hydroxyl group of chitosan to give 6- O -trityl chitosan has also been utilized to facilitate the synthesis of N -chloroacyl [16,17], N -betaine [18] and quaternary piperazine derivatives of chitosan [19,20]. Although the use of such selective protection resulted in higher DS, this led to an increase in the number of synthetic steps, and some modification at unprotected hydroxyl groups can also be observed [21]. Recently, we reported on silyl protected 3,6-di- O - tert -butyldimethylsilylchitosan (diTBDMS-CS) [22,23], which has been utilized in various chemoselective modifications to give products like N -(bromoacetyl)-diTBDMS-chitosan, N -(2-( N , N , N -trialkylammoniumyl)-chitosan, N , N , N -trimethyl chitosan and chitosan derivatives modified by covalent linking of the highly lipophilic photosensitizer, meso -tetraphenylporphyrin [24,25]. The TBDMS-protected precursor enabled the synthesis to be carried out in an organic medium, thereby allowing well controlled and regioselective modification, leading to homogenous products that can be fully characterized by spectroscopy with techniques, such as 1 H-NMR, FT-IR, COSY and HSQC. The role of the cationic charge in the antimicrobial effect is believed to be associated with the binding of the polymer to the bacterial cell wall. Several models have been proposed to explain the antimicrobial activity of chitosan, but the most accepted is electrostatic interaction between the positive charges on the polymer and the negatively charged anionic components of the bacterial surface, which weakens the cell wall and leads to cell lysis [26]. The polycationic structure of chitosan is a pre-requisite for antibacterial activity in spite of the structural differences in Gram positive and Gram negative bacteria [27]. Removal of the cell wall brings the polymer in contact with the cell membrane, thereby affecting membrane permeability and even reversing the surface charge of the bacteria [28]. These reactions finally lead to the leakage of the intracellular components, as evidenced by increased absorption at 260 nm [28], the increased electrical conductivity of the cell suspension [29] and cytoplasmic ȕ -galactosidase release [30–33]. The structure-activity relationship (SAR) for chitosan and chitosan derivatives is not well understood. The relation between molecular weight (Mw) and degree of acetylation (DA) of chitosan to its antibacterial properties has also been explored. While high Mw and degree of quaternization (DQ) of N , N , N -trimethyl chitosan chloride (TMC) derivatives showed high bactericidal activity against both S. aureus and E. coli [34], in another study, it was reported that low Mw chitosan and its derivatives showed better activity [35,36]. A lower DA of acid-soluble chitosan was shown to lead to a greater inhibitory effect against S. aureus and E. coli [37–39], while some other studies have not shown a clear relationship between DA and the antimicrobial effect of unmodified chitosan [40,41]. 5 In the current study, we used five different parent chitosan materials with variations in DA and Mw. These materials were used to synthesize different N -modified alkyl quaternary ammoniumyl and pyridiniumyl chitosan derivatives, such as (trimethylammoniumyl)acetyl, (trimethylammoniumyl)hexanoyl, (1-pyridiniumyl)acetyl, (1-pyridiniumyl)hexanoyl and N , N , N -trimethyl chitosan. These quaternary chitosan derivatives were then investigated for their antibacterial effects to allow systematic investigation of SAR under conditions where the effect of the functional group and the spacer length, as well as variations in the activity with the Mw and DA of the chitosan could be observed. 2. Results and Discussion The quaternary ammoniumyl and 1-pyridiniumyl derivatives were synthesized from five different chitosan parent materials (denoted in superscript, e.g., i – v ) (CS i – v ) varying in their DA from 6% to 34% and from 180 to 308 kDa in their Mw. The quaternary groups were distanced from the polymer backbone with alkyl chain spacers. Each spacer length required a different approach to the synthesis; the discussion on the synthesized derivatives is therefore divided into four sections in accordance with the length of the alkyl chain (C-spacer) or its absence. 2.1. Synthesis of N-(2-(N,N,N-Trimethylammoniumyl)acetyl)-chitosan Chloride (TMA-CS) and N-(2-(1-Pyridiniumyl)acetyl)-chitosan Chloride (PyA-CS), the C-2 Spacer Chitosan Derivatives The synthetic route to prepare the final TMA-CS ( 6 i – v ) and PyA-CS ( 8 i – v ) is shown in Scheme 1. Initially, all five different chitosan materials ( 1 i – v ) were converted to their corresponding chitosan mesylate salts (Mes-CS) ( 2 i – v ) by careful dropwise addition of methanesulfonic acid to the chitosan suspended in water at 10 °C. The finely powdered materials ( 2 i – v ) were obtained by following our earlier reported protocol. Unlike chitosan starting materials ( 1 i – v ), these mesylates, 2 i – v , were completely soluble in H 2 O, as well as in organic solvents, such as DMSO. The solubility of Mes-CS in DMSO was important, as it facilitated quantitative silyl protection of both hydroxyl groups on the CS under homogeneous conditions. Fully silyl-protected diTBDMS-CS ( 3 i – v ) materials were then obtained by using tert-butyl-dimethylsilyl chloride (TBDMSCl) and imidazole in DMSO at 25 °C. The intermediate N -(bromoacetyl)-3,6-di- O -TBDMS-chitosan (BrA-diTBDMS-CS) ( 4 i – v ) was prepared by reacting silyl chitosan 3 i – v with four equivalents of bromoacetyl bromide in the presence of five equivalents of triethylamine (Et 3 N). The reaction temperature was carefully maintained at í 20 °C throughout the reaction, and the reaction was quenched after 1 h to avoid any side reactions. The crude material was washed with acetonitrile (CH 3 CN) to afford the fine powdered material, which was completely soluble in dichloromethane (CH 2 Cl 2 ). Freshly prepared reactive intermediate 4 i – v was then reacted at 25 °C in CH 2 Cl 2 , with an excess of NMe 3 or pyridine to afford compounds 5 i – v and 7 i – v , respectively. Compounds 5 i – v and 7 i – v were finally deprotected using concentrated (conc) HCl/MeOH to afford the corresponding final quaternized chitosan derivatives, 6 i – v and 8 i – v , respectively (Scheme 1).