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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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Determination of oxidative stress biomarkers by HPLC
Váňová, Nela ; Jun, Daniel (advisor) ; Doležal, Rafael (referee) ; Hroch, Miloš (referee)
and keywords Although reactive oxygen and nitrogen species have a fundamental role in physiological processes occurring in living organisms, their overproduction induced by endogenous and/or exogenous sources may lead to serious imbalance in redox homeostasis, damage to intracellular components and thus, dramatically alter their function or even trigger cell death. Oxidative stress is believed to be very important mechanism of toxicity of xenobiotics, including drugs, and may be responsible for the development of their unwanted side effects. Considering a very low number of studies evaluating oxidative stress after the treatment with oxime reactivators of acetylcholinesterase (AChE) in vivo and in vitro, the relationship between their toxicity and generation of specific biomarkers of oxidative damage is not still fully understood. In order to monitor antioxidant/prooxidant properties of drugs, high performance liquid chromatography method coupled with tandem mass spectrometry (LC-MS/MS) for simultaneous determination of two biomarkers of oxidative stress, malondialdehyde (MDA) and 3-nitrotyrosine (3-NT), in biological matrices was developed. Validation of this LC-MS/MS method demonstrated the acceptable appreciable selectivity, accuracy, intra- and interday precission, and recovery of sample...
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A theological view of healing in Christianity and selected spiritual streams of the 20th century
Doležal, Rafael ; Bouma, David (advisor) ; Ventura, Václav (referee)
DOLEŽAL Rafael: A theological view of healing in Christianity and selected spiritual streams of the 20th century, Praha: Teologická fakulta Univerzity Karlovy, 2012. The subject of the diploma thesis consists in study of healing, which is seen from a position of fundamental theology. Attention is paid especially to those claiming to be classified as miraculous ones. A theologically seized recovery is then compared with the concept of healing in selected spiritual streams of the 20th century, belonging to the group New Age. This diploma thesis analyzes the characteristics of miraculous healing, and examines their significance for religion. Keywords: fundamental theology, miracle, healing, New Age, holistic health
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Quantitative structure-antimycobacterial acitivity relationships in the group of potential antituberculotics
Doležal, Rafael ; Waisser, Karel (advisor) ; Lehotay, Jozef (referee) ; Ponec, Robert (referee)
DOLEŽAL Rafael: Quantitative relationships between the structure and antimycobacterial activity of selected potential antituberculotics. Hradec Králové: Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 2008, 274 pp. Ph.D. Thesis. The Ph.D. thesis deals with analysis of quantitative relationships between the structure and antimycobacterial activity (QSAR) of four structural classes of potential antituberculotics - derivatives of N-phenylcarbamic acid esters, derivatives of 1-phenyl-1H-tetrazole-5-thiol, derivatives of salicylamides and derivatives of salicylthioamides. In total, 600 computer models of different compounds were created. 204 of them, after conformational analysis in HyperChem 7.52 and geometric optimization with the B3LYP/6-31G* DFT method in Gaussian 03, provided data for calculation of several types of descriptors. Attention was turned especially to local, quantum-chemical descriptors of electronic properties, such as superdelocalizability, index of frontier electron density and Mulliken electric charge. The QSAR analyses are based on searching for statistically significant correlations between the antimycobacterial activities in vitro and a matrix of various descriptors by means of multiple linear regression (MLR) with the use of a selection algorithm, by the...
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