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Studying the distribution of substituents in derivatives based on hyaluronane
Sedláček, Jakub ; Šatínský, Dalibor (advisor) ; Zimčík, Petr (referee) ; Chmelař, Josef (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate Ing. Jakub Sedláček Supervisor doc. RNDr. Dalibor Šatínský, Ph.D. Title of Doctoral Thesis Studying the distribution of substituents in derivatives based on hyaluronan Derivatives based on hyaluronan have found applications in various areas of medicine. Prepared derivatives are commonly characterized by their molar mass and their degree of substitution, however, such parameters are unable to distinguish among derivatives with different distributions of substituents along the chain, which may exhibit different macroscopic properties. For this reason, methods to determine the distribution of substituents along the chain are needed. The presented thesis deals with the determination of distribution of amino groups in deacetylated hyaluronan. The theoretical part starts with a description of hyaluronan structure and of its derivatives. Next, the application, the preparation, and the characterization of deacetylated hyaluronan are discussed, focusing particularly on its depolymerisation into oligosaccharides using deaminative cleavage or enzymes that degrade hyaluronan. Finally, examples of liquid chromatographic methods used to analyse hyaluronan oligosaccharide are presented. The experimental part...
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Development and optimization of chromatographic methods suitable for clinical application
Jenčo, Jaroslav ; Solich, Petr (advisor) ; Hroboňová, Katarína (referee) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of Analytical Chemistry Candidate: Mgr. Jaroslav Jenčo Supervisor: prof. RNDr. Petr Solich, CSc. Consultant: doc. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of dissertation thesis: Development and optimization of chromatographic methods suitable for clinical application The presented dissertation thesis is divided into several parts, copying the workplaces, which I gradually worked on. The unifying element is the development and optimization of methods for the preparation of samples of biologically active substances and their subsequent analysis using separation methods. The theoretical part characterizes the target substances, from a biochemical, physicochemical, and analytical point of view. It describes the basic characteristics of extraction procedures and presents an overview of the methods used to determine the substances described in the thesis. In the first, main experimental part, of the dissertation thesis the development, optimization, and validation of an analytical method for chromatographic analysis of vitamin B1, its biologically active forms and the active form of vitamin B6 in blood is described. It illustrates the selection of sample preparation techniques, the optimization of the target methodology...
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Development of on-line SPE HPLC method for determination of ochratoxin A in Tokaj wines
Uhrová, Adéla ; Šatínský, Dalibor (advisor) ; Chocholouš, Petr (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Adéla Uhrová Supervisor, consultant: Doc. RNDr. Dalibor Šatinský, Ph.D. Title of Diploma Thesis: Development of on-line SPE HPLC method for determination of ochratoxin A in Tokaj wines This diploma thesis deals with the optimization and validation of chromatographic method for determination of ochratoxin A in Tokaj wine with subsequent application on real samples. A high- performance liquid chromatography method with fluorescence detection using on-line extraction by column switching has been developed. 50µl of sample was injected directly in the extraction column, where at 1st minute the analyte was extracted on the Ascentis® Express RP-Amide precolumn in the presence of a mobile phase of 40% MeOH in 0.5% aqueous acetic acid. After the extraction, the valve was switched and the extracted substances were separated on a YMC Triart C18 ExRS analytical column. The separation was carried out under gradient elution with an initial acetonitrile ratio of 45% in the mobile phase, which reached a concentration of 80% in the 5th minute. Fluorimeter detection was optimized for excitation and emission wavelengths ʎEx 335 nm and ʎEm 463 nm. In the last part of the work, ochratoxin A was analyzed in real...
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Development of HPLC method for determination of selected phenolic acids and flavonoids in Tokaj wines
Moravcová, Pavlína ; Šatínský, Dalibor (advisor) ; Lhotská, Ivona (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical chemistry Candidate: Bc. Pavlína Moravcová Supervisor: Doc. RNDr. Dalibor Šatínský, Ph.D. Title of the diploma thesis: Development of HPLC method for determination of selected phenolic acids and flavonoids in Tokaj wines The aim of this diploma thesis was to develop and optimize HPLC method for simultaneous determination of 15 phenolic compounds, protocatechic acid, fertaric, coffee, p-hydroxybenzoic, syringic, gallic, p-coumaric, ferulic, chlorogenic, caftaric, 2-hydroxy-4-methoxybenzoic , polydatin, resveratrol, catechin hydrate and epicatechin in Tokaj wines. The quantity and content of selected phenolic compounds was analyzed in 25 samples of Tokaj wines, which come from vineyards from the Slovak part of Tokaj wine region. The work was also focused on the description of the specific characteristics of this unique area, the unique method of Tokaj wine production, its chemical composition, and namely the spectrum of phenolic substances. Analysis of selected phenolic compounds was performed using an Ascentis® Express F5 column (150 x 4.6 mm; 5 µm) using a mobile phase consisting of 0.1% phosphoric acid and acetonitrile in gradient elution. The substances were analyzed at 30 řC and flow rate of 1 ml/min and detected...
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Optimization of neonicotinoids extraction using nanofibers in sequential injection analysis systém
Zouharová, Monika ; Horstkotte Šrámková, Ivana (advisor) ; Šatínský, Dalibor (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Monika Zouharová Supervisor: PharmDr. Ivana Horstkotte Šrámková, Ph.D. Title: Optimization of neonicotinoids extraction using nanofibers in sequential injection analysis system In this diploma thesis, we tested the use of various nanofibers as sorbents for solid phase extraction of neonicotinoids connected with sequential injection analysis system. Seven types of nanofibers were tested, specifically polyamide (PA6), polycaprolactone (PCL), polyimide (PID) and polydopamine coated polyamide (PA6)coated, polycaprolactone (PCL)coated, polyimide (PID)coated and a combination of polyimide and polycaprolactone (PID/PCL)coated. These nanofibers were produced by electrospinning. The studied nanofibers were placed in the 3D printed holder, which was connected into sequential injection analysis system. UV spectrophotometry was used as a detection method. The substances to analyse were neonicotinoid insectisides. The retention of these analytes was examinated with all seven types of nanofibers. Retention of the analytes on the tested nanofibers depending on their chemical structures and physical-chemical properties was studied in these measurements. The most experiments were performed with polydopamine coated...
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Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques
Javorská, Lenka ; Solich, Petr (advisor) ; Pohanka, Miroslav (referee) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of Analytical Chemistry Candidate: Mgr. Lenka Javorská Supervisor: prof. RNDr. Petr Solich, CSc. Consultant: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of dissertation thesis: Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques The presented dissertation is focused on the development and validation of chromatographic method for the determination of glycopeptide antibiotic vancomycin, and L-arginine and its pathway related substances in biofluids. Furthermore, the potential of neopterin, kynurenine and tryptophan as biomarkers of immune response to surgical procedure and prediction of postoperative complications is investigated in connection with oncological diseases. In the theoretical part trends in instrumentation, stationary phases and extraction techniques are described. The issues of separation and retention of polar compounds and biological matrix pretreatment before analysis are also discussed there. Next part deals with selected analytes and their application to the clinical purpose for which the chromatographic methods are developed. It also provides an overview of published methods for their determination using liquid...
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Application of nanofibrous sorbents for on-line extraction in liquid chromatography
Háková, Martina ; Šatínský, Dalibor (advisor) ; Česla, Petr (referee) ; Urban, Jiří (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Martina Háková Supervisor: doc. RNDr. Dalibor Šatínský, Ph.D. Consultant: doc. PharmDr. Lucie Chocholoušová Havlíková, Ph.D. Title of Doctoral Thesis: Application of nanofibrous sorbents for on-line extraction in liquid chromatography A compilation of ten publications is presented in this dissertation thesis. It is focused on the application of nano and microfibrous polymers as a new extracting material in analytical chemistry. This work was based on close cooperation with the Institute for Nanomaterials, Advanced Technologies and Innovation and the Department of Nonwovens and Nanofibrous Materials at the Technical University of Liberec, where the author took part in designing and preparation of these materials. The first part deals with model studies, based on using polyamide, polystyrene and polycaprolactone nanofibers as sorbents for solid phase extraction in classical off-line mode or in on-line column switching system. It solves practical aspects such as orientation and the formation of the fibers in the extraction system, optimization of fabrication factors affecting applicability of the nanomaterials, suitable solvent systems, stability testing and reusability of the microcolumns. The...
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Evaluation of selected substances in fruit by HPLC-DAD-CAD method
Košková, Stanislava ; Sklenářová, Hana (advisor) ; Šatínský, Dalibor (referee)
Charles University Faculty of Pharmacy in Hradci Králové Deparment of Analytical Chemistry Candidate: Bc. Stanislava Košková Supervisor: Doc. PharmDr. Hana Sklenářová, Ph.D. Consultant: Ing. Radek Vávra, Ph.D. (VŠÚO Holovousy) Title of diploma thesis: Evaluation of selected substances in fruit by HPLC-DAD-CAD method Thisdiploma thesis was focused on determination of seven phenolic compounds, namely gallic acid, chlorogenic acid, epicatechin, rutin, phloridzin, quercetin, phloretin in twenty apple cultivars, that have been delivered by the Research and Breeding Institute of Pomology Holovousy s.r.o. For this purpose HPLC separation in reverse mode was used, using the column Omega Polar C 18 (150 mm x 4.6 mm; 5 µm). As the mobile phase the mixture of an ultrapure water acidified with the acetic acid to pH 2.8 and acetonitrile was used. A linear gradient was chosen with a gradual increase of the organic component from the initial 10 % to 50 % during 10 min of separation.This was followed with column equilibration - up to 12.50 min to 10 % organic component. The overall separation time was 12.50 min at a flow rate of 1 ml/min. Column temperature was set to 30 řC. Detection was performed using DAD (254, 280, 320 nm) and CAD detector (nebulizer temperature 25 řC). Part of this thesis is system...
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Comparison of methods for determination of amphetamine derivates in meconium
Vrbová, Romana ; Matysová, Ludmila (advisor) ; Šatínský, Dalibor (referee)
Department: Department of Analytical Chemistry Candidate: Romana Vrbová Supervisor: doc. PharmDr. Ludmila Matysová, Ph.D. Title of the diploma thesis: Comparison of methods for determination of amphetamine derivates in meconium The aim of the bachelor thesis was to optimize a new method for the determination of amphetamine derivatives in meconium, which will be faster and simpler than the existing routine method in the toxicological laboratory of Masaryk's Hospital in Ústí nad Labem. The theoretical part of the bachelor thesis describes the main representatives of amphetamine derivatives, their effects and metabolism. Their determination in meconium is important for the diagnosis of Neonatal abstinence syndrome, which occurs in newborns of mothers using addictive substances during pregnancy. Meconium, the first stool of a newborn, is a suitable material demonstrating exposure to drugs and medication during the second and third trimesters of pregnancy. Amphetamine derivatives were determined in meconium by gas chromatography with mass detection, the principles and instrumentalization of which are another part of the bachelor's thesis. The experimental part describes the optimization of a new, more advantageous method using pentafluorobenzoyl chloride as a derivatizing agent and its comparison with...
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Development of CE-C4D method for the analysis of saccharides in honey
Šebestová, Gabriela ; Jáč, Pavel (advisor) ; Šatínský, Dalibor (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Gabriela Šebestová Supervisor: PharmDr. Pavel Jáč, Ph.D. Title of the diploma thesis: Development of CE-C4D method for the analysis of saccharides in honey Honey is a commonly used natural mixture which quality must be monitored due to possible mishandling or adulteration. This diploma thesis is focused on the development of a capillary electrophoresis method with capacitively coupled contactless conductivity detection (CE-C4D) for the determination of carbohydrates in honey samples. Design of Experiments (DoE) was used to optimize the separation conditions. This approach allows to find suitable separation conditions by means of multivariate optimization. During the optimization, the impact of the following factors was investigated: the composition of the background electrolyte, the separation voltage and temperature during separation and the inner diameter of the separation capillary. Two methods were optimized: one with normal electroosmotic flow (EOF) and the other using reversed EOF after CTAB addition. The method in the normal EOF mode was used for validation and analysis of honey samples. A fused silica capillary with an inner diameter of 20 m, a total length of 48.7 cm and an effective...
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