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LC-HRMS analysis of selected drugs in biological material II.
Švarcová, Kristýna ; Kučera, Radim (advisor) ; Bavlovič Piskáčková, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Kristýna Švarcová Consultant: RNDr. Martin Mžik, Ph.D. Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Title: LC-HRMS analysis of selected drugs in biological material II. Anxiety disorders affect up to 25 % of the population in their life, the lifetime prevalence is 13,6 %. Insomnia affect at least one third of the population in their life, the prevalence is 15-40 %. Drugs used to treat anxiety and insomnia easily cross the blood-brain barrier and affect the CNS. It is therefore necessary to determine the correct dosing schedule and monitor treatment. The result is optimization of treatment, reduction of side effects and increased patient adherence to treatment. The rapid qualitative and quantitative analysis for urgent cases of intoxication is also needed. This diploma thesis deals with development and optimization of an extraction method for selected benzodiazepines (alprazolam, bromazepam, diazepam, chlordiazepoxide, clonazepam, midazolam, oxazepam) and zolpidem in human serum. Subsequently, the whole analytical method was validated for use in clinical practice using the UHPLC-HRMS. Protein precipitation with acetonitrile was the most suitable sample preparation...
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The influence of solubility and adsorption on plastic materials on transport experiments
Šilhanová, Marie ; Kučera, Radim (advisor) ; Váňová, Nela (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department: Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Student: Marie Šilhanová Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Title of diploma thesis: The influence of solubility and adsorption on plastic materials on transport experiments From transport experiments on cell culture models we get valuable information about transport mechanism of drugs in organism. In vitro experiments are conducted for example on Transwell type inserts. During the experiment it was discovered that the results are not homogeneous, and the quantity of a substance in the solution decreases apparently, the reason behind this is inadequate solubility of lipophilic substances or their adsorption on the surface of plastic materials used in the experiment. Due to these problems we experience significant bias. This thesis is focused on antivirotics that did not perform well during transport experiments. First, HPLC/MS methods were developed, and they were used for concentration measurement of samples containing individual antivirotics. The drugs were tested under wide range of conditions so possible changes in effects of adsorption on plastic surfaces and solubility of drugs could be observed. The substances were divided into groups based on...
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The electrochemical reduction of the nitro group in a flow reactor
Jankových, Miroslav ; Kučera, Radim (advisor) ; Zimčík, Petr (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical chemistry and Pharmaceutical analysis Student: Miroslav Jankových Supervisor: doc. PharmDr. Radim Kučera, PhD. Consultant: PharmDr. Tomáš Holas, PhD. Title of diploma thesis: The electrochemical reduction of the nitro group in a flow reactor The aim of this diploma thesis is to study the electrochemical reduction within the flow reactor. The combination of these two methods provides enviromentaly-friendly and interesting method for the manufacturing of active pharmaceutical ingredients. Our object was to find out if the reaction really goes on. After that our goals were to optimize the reaction parameters. We executed some of the experiments to understand the parameters such as: flow rate, voltage, electric current, electrolyte. These parameters we combined to see their effect on the reaction results. The outcome of the thesis is mainly basic information about impact of parameters mentioned above. From the series of experiments the best parameters are flow rate 0,1 ml.min- 1 , voltage 0,8 - 2,0 V, electric current 100 - 150 mA, electrolyte NH4Cl, reaching conversion up to 96%.
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The employment of HPLC for separation of bioconjugates of azaphthalocyanines II.
Sčensná, Anna ; Kučera, Radim (advisor) ; Lochman, Lukáš (referee)
7 ABSTRACT Charles University, Faculty of Pharmacy in Hradec Králové Department Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate Anna Sčensná Supervisor doc. PharmDr. Radim Kučera, Ph.D. Title of Thesis The employment of HPLC for separation of bioconjugates of azaphthalocyanines II. Bioconjugates of azaphthalocyanines are studied as dark quenchers in molecular probes to monitor real-time PCR. The detection is based on energy transfer between a fluorophore and a quencher, both labelled to the oligonucleotide chain. The fluorophore emits a radiation and the dark quencher switches it off. The efficiency of fluorescence quenching depends on the distance between fluorophore and quencher. Probes can be mono-labelled, in which two oligonucleotides are used, one attached to a fluorophore and the other one to a quencher. The second type is called double-labelled probes, which contain both (fluorophore and quencher) on the same oligonucleotide chain. An unbound probe is randomly coiled, thus fluorophore and quencher approach each other. The fluorescence is not emitted. After hybridisation with a complementary chain, the quencher and fluorophore move away from each other. The terminal nukleosid at 5'end of Taq-man probe is cleaved after hybridisation, where the fluorophore is bonded. After...
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Usage of HPLC and UHPLC-MS/MS techniques for clinical research
Kučerová, Kateřina ; Matysová, Ludmila (advisor) ; Doležal, Rafael (referee) ; Kučera, Radim (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Kateřina Kučerová Supervisor: assoc. prof. PharmDr. Ludmila Matysová, Ph.D. Consultant: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of Dissertation Thesis: Usage of HPLC and UHPLC - MS/MS techniques for clinical research The dissertation thesis is dealing with the extraction techniques and with development, optimization and validation of chromatographic methods for the determination of omeprazol which is drug from the group of proton pump inhibitors, and retinol as a new potential biomarker of kidney damage. The theoretical part of the thesis is focused on detailed description of the individual analytes, their clinical significance, options of determination and the matrix in which the target analytes were determined. Furthermore, the preparation of the sample before the analysis is discussed and validation recommendations related to chromatographic methods used in both pharmaceutical analysis and bioanalysis are also mentioned. In the experimental part the development, optimization and validation of UHPLC methods and their subsequent use are described in detail. The pharmaceutical-analytical part of the work describes the determination of omeprazole in new suspension preparations,...
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The development of new methods of ultra-high performance supercritical fluid chromatography for pharmaceutical applications
Plachká, Kateřina ; Nováková, Lucie (advisor) ; Kučera, Radim (referee) ; Novák, Ondřej (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Kateřina Plachká Supervisor: prof. Pharm.Dr. Lucie Nováková, Ph.D. Title of Doctoral Thesis: The development of new methods of ultra-high performance supercritical fluid chromatography for pharmaceutical applications A compilation of seven articles and three book chapters dealing with the applicability of supercritical fluid chromatography (SFC) methods in the field of pharmaceutical analysis is presented in this dissertation thesis. The first part focuses on pharmaceutical quality control and especially impurity control. The presented articles are dealing with the searching for generic screening approach for the development of SFC methods and with the subsequent optimization and validation of these methods. The screening conditions were compared based on developed mathematical model. The generic combination of diol stationary phase and 0.1 % ammonium hydroxide in methanol as modifier of CO2-based mobile phase was proposed as a starting point for SFC method development based on the obtained results. Several challenges were described during optimization of UHPSFC methods and their solution was proposed. The benefits of acetonitrile added to the modifier, advantages of column coupling and the...
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The use of HPLC in the field of chiral separations IV.
Daněk, Alois ; Kučera, Radim (advisor) ; Štěrbová, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Alois Daněk Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Title of thesis: The employment of HPLC in the field od chiral separations IV. The requirements for the quality and safety of medicines are constantly growing at present. Drugs are often chiral substances. Therefore, the importance of chiral separations is increasing. Direct separations using chiral stationary phases are the most used in this field. Silica gel, which is most often used as a carrier for chiral selectors, may contain metal impurities on its surface, which may negatively affect the separation process. This thesis focuses on testing the influence of chromatographic conditions on chiral and achiral interactions of nine selected analytes with the stationary phase. A column with β-cyclodextrin as chiral selector was used for the testing.
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Strategie rozvoje společnosti VINOK s.r.o.
Kučera, Radim
The bachelor thesis describes the strategic analysis of VINOK s.r.o. company using analysis methods of internal and external factors. Selected key factors are used for creating strategic proposals, one of the proposals is described in detail.
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Development of HPLC method for evaluation of tryptophan metabolites
Pšenčíková, Barbora ; Kastner, Petr (advisor) ; Kučera, Radim (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Barbora Pšenčíková Supervisor: PharmDr. Petr Kastner, Ph.D. Title of thesis: Development of HPLC method for evaluation of tryptophan metabolites The essential amino acid tryptophan is a precursor of several important bioactive compounds. In recent years it was found out that tryptophan metabolites are involved in the pathogenesis of many diseases, including neurodegenerative and psychiatric disorders, autoimmune diseases, or cancer. Therefore, the analytical methods for simultaneous determination of tryptophan and its metabolites are highly required. The purpose of this diploma thesis was to develop an optimal HPLC method for determination of tryptophan and its metabolites (kynurenine, kynurenic acid, serotonin and 5-hydroxyindole-3-acetic acid) and to consider all the possible options of determination of quinolinic acid and melatonin. The chromatographic separation was carried out by Supelco Ascentis Express F5 column, 2.7 μm particle size, 15 cm × 3 mm, using spectrophotometric and fluorescence detection. Parameters of detection were for kynurenine UV detection 369 nm, 254 nm, fluorescence detection Ex/Em: 369/475; for kynurenic acid UV detection 244 nm; for...
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LC-HRMS analysis of selected drugs in biological material I.
Tomanová, Jana ; Kučera, Radim (advisor) ; Váňová, Nela (referee)
Charles Univesity, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Jana Tomanová Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Consultant: PharmDr. Vít Šesták, Ph.D. Title of the Diploma Thesis: LC-HRMS analysis of selected drugs in biological material I. High performance liquid chromatography is one of the most widespread separation analytical techniques. It is used in various medical, pharmaceutical and industrial laboratories. It uses different affinities of the substances in the analysed mixture to the mobile and stationary phases. Mass spectrometry is one of the instrumental techniques by which charged particles are separated in the gas phase based on the mass- to-charge ratio. Nowadays, great emphasis is placed on the fact that the individual analytical methods which are being developed are subsequently validated according to a guideline issued by national or international authorities. Validation is a process that verifies that a method is suitable for its intended use. In this thesis, the conditions for the determination of warfarin in patient's serum by means of mass spectrometer with LTQ XL linear ion trap were optimized and validated. The optimization of the method was based on the already developed and validated...
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