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Optimization of preparative LC-MS method for fractionation of oligosaccharides of hyaluronan
Dvořáková, Martina ; Lázníčková, Alice (advisor) ; Čožíková, Dagmar (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biophysic and Physical Chemistry Candidate: Bc. Martina Dvořáková Supervisor: Doc. Ing. Alice Lázníčková, CSc. Consultant: Mgr. Martina Hermannová, Ph.D. Title of diploma thesis: Optimization of preparative LC-MS method for fractionation of oligosaccharides of hyaluronanu This diploma thesis deals with optimization of LC-MS method for analysis of hyaluronan oligosaccharides in preparative mode. The theoretical part summarizes available information about biological and chemical properties of hyaluronic acid. Hyaluronic acid is easily enzymatically degradable by mammalian hyaluronidases that produce hyaluronan oligosaccharides. The biological function of these degradation products depend on their molecular weight. High-performance liquid chromatography is mainly used for separation and purification of hyaluronan oligosaccharides. A new method for the determination of hyaluronan oligosaccharides is based on a combination of separation techniques and mass spectrometry. The experimental part deals with optimization of ionisation conditions for electrospray ionization mass spectrometry in positive and negative ion mode. In the first step, we focused on setting of capillary voltage, cone voltage, desolvation temperature, flow...
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Analysis of Radiolabeled Hyaluronic Acid and its Metabolites in Biological Material
Čožíková, Dagmar ; Lázníčková, Alice (advisor) ; Kubala, Lukáš (referee) ; Hrdina, Radomír (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate Ing. Dagmar Čožíková Supervisor Doc. Ing. Alice Lázníčková, CSc. Title of Doctoral Thesis Analysis of radiolabeled hyaluronic acid and its metabolites in biological material The presented dissertation thesis deals with an optimization process of radiolabeled techniques and the following analysis of radioactive HA in biological material. Theoretical part include basic information about physical, chemical and biological properties of HA, and its function and degradation processes in vitro and in vivo. Furthermore, there are described analytical methods for the determination of HA, and also some of the most employed radiolabeled techniques of HA and its derivatives. Last but not least, there are summarized all the latest knowledge about biodistribution and pharmacokinetics of HA after intravenous, or topical administration, etc. In addition, we can also find there the most often medical and cosmetic application of HA. Radiolabeled techniques using four different radionuclides (111In, 125I, 14C and 99mTc) were developed: chemical synthesis, biosynthesis, and direct labeling method. Consequently, purification process of radiolabeled HA using separation on sephadex G-50 or...
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