|
|
Application of liquid chromatography in the analysis of biological material for clinical research
Turoňová, Dorota ; Kujovská Krčmová, Lenka (advisor) ; Pohanka, Miroslav (referee) ; Solichová, Dagmar (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Dorota Turoňová Supervisor: Assoc.prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of doctoral thesis: Application of liquid chromatography in the analysis of biological material for clinical research The dissertation focuses on the use of liquid chromatography for clinical research. It is divided into two main sections, Theoretical part and Commentary on selected published articles. The theoretical part describes the application of high-performance liquid chromatography in practice. Key parameters for the development of LC methods and trends in current bioanalytical practice are discussed. Furthermore, the sample collection and preparation of different biological matrices is explained, including all the obstacles and pitfalls that need to be taken into account. Modern trends are also briefly mentioned. The chapter on sample preparation also includes a section on derivatization and its use in the analysis of platinum derivatives, which is related to the commentary on the published work. The main part of the thesis deals with commentary on four published articles. The first presents a review article dealing with the use of microextraction in pipette tips in clinical and forensic toxicology....
|
|
|
Application of liquid chromatography in the analysis of biological material for clinical research
Turoňová, Dorota ; Kujovská Krčmová, Lenka (advisor) ; Pohanka, Miroslav (referee) ; Solichová, Dagmar (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Dorota Turoňová Supervisor: Assoc.prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of doctoral thesis: Application of liquid chromatography in the analysis of biological material for clinical research The dissertation focuses on the use of liquid chromatography for clinical research. It is divided into two main sections, Theoretical part and Commentary on selected published articles. The theoretical part describes the application of high-performance liquid chromatography in practice. Key parameters for the development of LC methods and trends in current bioanalytical practice are discussed. Furthermore, the sample collection and preparation of different biological matrices is explained, including all the obstacles and pitfalls that need to be taken into account. Modern trends are also briefly mentioned. The chapter on sample preparation also includes a section on derivatization and its use in the analysis of platinum derivatives, which is related to the commentary on the published work. The main part of the thesis deals with commentary on four published articles. The first presents a review article dealing with the use of microextraction in pipette tips in clinical and forensic toxicology....
|
|
|
The Impact of Metabolic Syndrome on the Expression of Inflammation and Cellular Damage Parameters in Relation to the Presence of Psoriasis
Holmannová, Drahomíra ; Fiala, Zdeněk (advisor) ; Dlouhý, Pavel (referee) ; Kujovská Krčmová, Lenka (referee)
Metabolic syndrome is a chronic metabolic and systemic inflammatory disease that is associated with high morbidity and mortality. The increasing prevalence of metabolic syndrome highlights the need for a deeper understanding of the pathophysiological mechanisms and their interrelationship with common comorbidities, including psoriasis. By analysing changes in the levels of selected parameters of inflammation and cellular damage, the knowledge of the pathogenesis of the inflammatory process of both diseases and their combination can be expanded. The results may contribute to effective secondary prevention and primary care interventions and facilitate modification of therapies. Data for the doctoral thesis were obtained from three separate studies, for which a set of parameters reflecting metabolic status, inflammatory processes, and cell damage (oxidative DNA/RNA damage, calprotectin, angiopoietin-like protein 8 /ANGPTL8/, clusterin, elafin, chromosomal aberrations, CRP, and vitamin D) were selected. The studies included groups of people with metabolic syndrome, persons with psoriasis, persons with psoriasis and metabolic syndrome, and healthy subjects. A total of 248 subjects participated in the study in three studies (122, 85 and 41 subjects). Laboratory determination of parameters was performed by...
|
|
|
Control of mycotoxin content in food supplements
Přívratská, Nikola ; Šatínský, Dalibor (advisor) ; Kujovská Krčmová, Lenka (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Nikola Přívratská Supervisor: prof. RNDr. Dalibor Šatínský, Ph.D. Title of the diploma thesis: Control of mycotoxin content in food supplements Due to the growing popularity of nutraceuticals, some concerns about their quality and safety have been expressed. The presented work describes the development of a fast method for mycotoxin ochratoxin A (OTA) determination using high performance liquid chromatography (HPLC) and column switching technique in combination with on-line solid phase extraction (SPE). The method was used for direct determination of OTA in milk thistle-based nutraceuticals. Nutraceuticals were extracted by methanol - water solution in a ratio of 1:9. A volume of 50 μl of the extracted sample was injected on the extraction column Ascentis Express RP-Amide (5 x 2.1 mm, 5 µm particle size). The sample was cleaned from interfering compounds with 20% methanol at a flow rate of 1 ml/min. After a period of 1 minute the valve was switched and mobile phase eluted OTA trapped on the extraction column to the analytical column. Separation was performed by gradient elution on the Ascentis Express Phenyl-Hexyl (100 x 4.6 mm, 5 µm particle size) column using a mobile phase consisting of...
|
|
|
Application of modern analytical methods in clinical research
Vernerová, Andrea ; Kujovská Krčmová, Lenka (advisor) ; Doležal, Rafael (referee) ; Malíř, František (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Training Workplace Department of Analytical Chemistry Doctoral Degree Program Bioanalytical Methods Candidate Mgr. Andrea Vernerová Supervisor Assoc. Prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Advisor MUDr. Ondřej Sobotka, Ph.D. Title of Doctoral Thesis Application of modern analytical methods in clinical research The dissertation thesis, composed as a compilation of five publications, is dealing with the development, optimization and validation of chromatographic methods and extraction procedures for the determination of early activation of the immune system biomarkers (neopterin, kynurenine, and tryptophan) in saliva and gingival crevicular fluid in oncological patients and patients with periodontal diseases. Furthermore, the potential of the use of Clark's sensor to determine the correct function of platelets by measuring mitochondrial respiration and its application in clinical research is under investigation. This work was based on cooperation with several clinical departments of the University Hospital Hradec Králové and Olomouc, and Oroboros Instruments (Innsbruck, Austria). The theoretical part of the thesis is focused on detail description of the individual analytes, their clinical significance, used biological matrices and their...
|
|
|
Application of modern analytical methods in clinical research
Vernerová, Andrea ; Kujovská Krčmová, Lenka (advisor) ; Doležal, Rafael (referee) ; Malíř, František (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Training Workplace Department of Analytical Chemistry Doctoral Degree Program Bioanalytical Methods Candidate Mgr. Andrea Vernerová Supervisor Assoc. Prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Advisor MUDr. Ondřej Sobotka, Ph.D. Title of Doctoral Thesis Application of modern analytical methods in clinical research The dissertation thesis, composed as a compilation of five publications, is dealing with the development, optimization and validation of chromatographic methods and extraction procedures for the determination of early activation of the immune system biomarkers (neopterin, kynurenine, and tryptophan) in saliva and gingival crevicular fluid in oncological patients and patients with periodontal diseases. Furthermore, the potential of the use of Clark's sensor to determine the correct function of platelets by measuring mitochondrial respiration and its application in clinical research is under investigation. This work was based on cooperation with several clinical departments of the University Hospital Hradec Králové and Olomouc, and Oroboros Instruments (Innsbruck, Austria). The theoretical part of the thesis is focused on detail description of the individual analytes, their clinical significance, used biological matrices and their...
|
|
|
Development and validation of UHPLC-MS/MS method for determination of urea in exhalted breath condensate and its application in clinical research
Vrbová, Lenka ; Kujovská Krčmová, Lenka (advisor) ; Matysová, Ludmila (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Lenka Vrbová Supervisor: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Consultant: PharmDr. Kateřina Matoušová, Ph.D. Title of the diploma thesis: Development and validation of UHPLC-MS/MS method for determination of urea in exhaled breath condensate and its application in clinical research Key words: urea, exhaled breath condensate, UHPLC-MS/MS Exhaled breath condensate analysis is a simple non-invasive method of measuring biomarkers not only from the respiratory tract. For the proper clinical interpretation of the measured data it is necessary to find the dilution biomarker due to dilution of target analytes with condensed water vapour. Urea seems to be the molecule of choice. The aim of this diploma thesis was to develop a fast, simple and sensitive analytical method for determining urea concentration levels in exhaled breath condensate for clinical research. Ultra-high performance liquid chromatography with mass spectrometry detection was chosen for urea analysis. Kinetex HILIC 100 A column, 1.7 µm particle size, 100 × 2.10 mm (Phenomenex, USA) was used for the separation. The mobile phase was composed of acetonitrile, methanol and water in a ratio of 81 : 9 : 10, the organic part was...
|
|
|
Development of HPLC method for determination of amiloride hydrochloride in pharmaceutical preparation
Sohrová, Barbora ; Matysová, Ludmila (advisor) ; Kujovská Krčmová, Lenka (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Barbora Sohrová Consultant: PharmDr. Ludmila Matysová, Ph.D. Diploma Thesis Title: Development of HPLC method for the determination of amiloride hydrochloride in medicinal preparation The purpose of thesis is the development of HPLC methods for determination of amiloride hydrochloride in pharmaceutical preparation. A medicinal product containing amiloride hydrochloride is used in the treatment of cystic fibrosis as a solution for inhalation. During the development, various stationary and mobile phases were tested. For the measurement was selected HS F5 column (150x 2,1 mm, 3 µm) and as the mobile phase was mixture of acetonitrile and 0,085% phosphoric acid to pH adjusted to 3,0 with triethylamine, in the ratio 75:25. Standard solution contained amiloride hydrochloride, Impurity A as a degradation product and butylparaben as an internal standard. The developed method was completely validated. According to results of the validation this method, was found that the developed method for the analysis of the product is suitable, and provides accurate and precise results. One of the findings of validation was the fact that the medicinal product containing amiloride hydrochloride is not stable...
|
|
|
Development of HPLC method for the determination of homatropine and scopolamine in eye drops
Sohrová, Barbora ; Matysová, Ludmila (advisor) ; Kujovská Krčmová, Lenka (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Barbora Sohrová Consultant: PharmDr. Ludmila Matysová, Ph.D. Thesis Title: Development of HPLC method for the determination of homatropine and scopolamine in eye drops High performance liquid chromatography (HPLC) method for the determination of homatropine hydrobromide and its degradation product- scopolamine hydrobromide in eye drops at a concentration of 1% (with sodium chloride), 2% (with sodium chloride) and 2% (with buffer F 6.45) active substance was developed. During the development of the method, various mobile and stationary phases were tested. For the validation was chosen column Ascentis Express F5 (100 x 4.6 mm, 2.7 µm). The mobile phase was composed of acetonitrile and phosphate buffer in a ratio of 9:91 (V/V). Phosphate buffer contained 3.12% aqueous solution of sodium dihydrogen phosphate dihydrate and 200 µl of triethylamine. Buffer pH was adjusted with phosphoric acid 85% to the value 2.5. Injection volume was 5 µl, flow rate was 1.1 ml/min and UV detection was performed at a wavelength of 210 nm. Analysis time was less than 6.5 minutes. Analyzed solution contained standard solution homatropine hydrobromide at a concentration of 25 mg/100 ml, and its degradation...
|
|
|
Development of new types of biocompatible hemodialysis membranes for separation of biomolecules
Kohlová, Michaela ; Solich, Petr (advisor) ; Kujovská Krčmová, Lenka (referee) ; Pena, Angelina (referee)
Charles University, Faculty of Pharmacy in Hradec Králové / University of Porto, Faculty of Pharmacy Department of Analytical Chemistry / Department of Chemical Sciences Candidate: Mgr. Michaela Kohlová Supervisor: prof. RNDr. Petr Solich, CSc. Consultants: prof. Maria de Conceição Branco Montenegro assoc. prof. Alice Santos-Silva Title of dissertation thesis: Development of new types of biocompatible haemodialysis membranes for separation of biomolecules This dissertation thesis was focused on the development, preparation, and optimization of flat sheet polysulfone membranes, which would meet the characteristics required for membranes used for haemodialysis treatment. The thin porous membranes were prepared by spin coating method, followed by phase inversion via immersion precipitation. The composition, as well as the preparation process of the membranes, were optimized, to obtain a membrane with good mechanical performance in flow conditions, adequate selective separation ability, and biocompatibility. The prepared membranes with the appropriate characteristics were modified with bioactive compounds to minimize oxidative stress and/or inflammation, which are common complications in haemodialysis treated patients. Three different modification approaches were used to prepare the bioactive...
|
guest ::
