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HPLC Determination of Neopterin, Tryptophan, Kynurenine and Creatinine in Human Serum
Prokopová, Veronika ; Solich, Petr (advisor) ; Matysová, Ludmila (referee)
In this work the new HPLC method for simultaneous determination of neopterin, tryptophan, kynurenine and creatinine was developed and validated. For separation of analytes two monolithic columns (Chromolith Speed ROD RP-18e, 50 x 4.6 mm and Chromolith Performance RP-18e, 100 x 3.0 mm) were connected together. In front of this connection Chromolith Guard RP-18e, 10 x 4.6 mm disc for better separation and serum proteins elimination was used. Neopterin and tryptophan were monitored using fluorescent detection. Neopterin was monitored at 353 nm excitation and 438 nm emission wavelength, for tryptophan 245 nm excitation and 404 emission wavelength was used. The detection of creatinine and kynurenine was carried out with help of diode array detector at wavelength 235 nm for creatinine and 230 nm for kynurenine. As the mobile phase 15 mmol/l phosphate buffer at pH 4.51 was used with the flow rate 1 ml/min in time 0 - 3 minutes, then the flow rate was changed to 2.3 ml/min. The total time of analysis was 9.5 minutes including the column equilibration. The injection volume of sample was 1 µl. This method was developed and validated using the standards of analytes and also in human serum.
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Application of novel stationary phases for HPLC determination of neopterine and other metabolites in urine.
Prokopová, Veronika ; Solich, Petr (advisor) ; Nováková, Lucie (referee)
The aim of this diploma thesis was to compare the separative options of different types of chromatographic columns for the determination of neopterin, kynurenine, creatinine and uric acid in urine samples with the method that is currently used in the Research laboratory of the Department of Methabolic Care and Gerontology, Teaching Hospital in Hradec Kralove. Basic, already developed and validated method was High Performance Liquid Chromatography (HPLC) on reversed-phase column Gemini 5 μm C18 110Ǻ, 150 x 3 mm, Phenomenex (Torrance, USA). Phosphate buffer with concentration of 15 mmol/l, pH 6,4 and flow rate 0,8 ml/min was used as a mobile phase. Diode array detector was used for detection of creatinine (235 nm ) , kynurenine (230 nm) and uric acid (290 nm) Neopterin was monitored using fluorescent detection at 353 nm excitation and 438 nm emission wavelength. All chromatographic systems were compared on the basis of the SST- System Suitability Test and were evaluated by the separation efficiency, peak resolution, asymmetry and other parameters. Columns with the best parameters were selected for further optimization and validation. The resulting new modern HPLC method should be suitable for clinical use.
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Bydlení v modulovém prostoru
Prokopová, Veronika
This thesis focuses on development of modular architecture and problematics of minimum housing. Findings are exploited in solving interior design of growth dispositional variations of modular housing. Besides design, esthetic or material processing of this modular space, it also deals with the design and the construction of atypical furniture element.
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HPLC Determination of Neopterin, Tryptophan, Kynurenine and Creatinine in Human Serum
Prokopová, Veronika ; Solich, Petr (advisor) ; Matysová, Ludmila (referee)
In this work the new HPLC method for simultaneous determination of neopterin, tryptophan, kynurenine and creatinine was developed and validated. For separation of analytes two monolithic columns (Chromolith Speed ROD RP-18e, 50 x 4.6 mm and Chromolith Performance RP-18e, 100 x 3.0 mm) were connected together. In front of this connection Chromolith Guard RP-18e, 10 x 4.6 mm disc for better separation and serum proteins elimination was used. Neopterin and tryptophan were monitored using fluorescent detection. Neopterin was monitored at 353 nm excitation and 438 nm emission wavelength, for tryptophan 245 nm excitation and 404 emission wavelength was used. The detection of creatinine and kynurenine was carried out with help of diode array detector at wavelength 235 nm for creatinine and 230 nm for kynurenine. As the mobile phase 15 mmol/l phosphate buffer at pH 4.51 was used with the flow rate 1 ml/min in time 0 - 3 minutes, then the flow rate was changed to 2.3 ml/min. The total time of analysis was 9.5 minutes including the column equilibration. The injection volume of sample was 1 µl. This method was developed and validated using the standards of analytes and also in human serum.
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The level of autoantibodies against procathepsin D in the different sets of patients
Prokopová, Veronika ; Dršata, Jaroslav (advisor) ; Springer, Drahomíra (referee)
VERONIKA PROKOPOVÁ THE LEVEL OF AUTOANTIBODIES AGAINST PROCATHEPSIN D IN THE DIFFERENT SETS OF PATIENTS ABSTRACT Cathepsin D (E.C. 3.4.23.5, CD) is lysosomal aspartic peptidase syntetized in all of the mammal cells and the ability to catalyze the hydrolitic cleavage peptidic bonds in peptides or proteins belongs to its basis characteristic. During the long-standing research it was discovered, that there exist an increased expression of CD in tumorous cells, which is more than 50 times higher than in normal cells and which leads to secretion of CD from these cells. Only enzymatically inactive zymogen procathepsin D (pCD) with intact activation peptide (AP) attached is being released by the tumorous cells. It was shown, that this activation peptide is probably responsible for the mitogenic activity in the tumorous cells. It comes up very often to the creation of antitumor immune response against tumor antigens at patients suffering from tumorous condition. Determination of these autoantibodies and analysing of diagnosed antigens could employed as diagnostic and prognostic markers. The aim of our study was to verify the hypothesis, that the level of autoantibodies against AP pCD could be employed for diagnostic purposes, first of all as a new tumorous marker. It was necessary to develop and optimalize ELISA...
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Application of novel stationary phases for HPLC determination of neopterine and other metabolites in urine.
Prokopová, Veronika ; Nováková, Lucie (referee) ; Solich, Petr (advisor)
The aim of this diploma thesis was to compare the separative options of different types of chromatographic columns for the determination of neopterin, kynurenine, creatinine and uric acid in urine samples with the method that is currently used in the Research laboratory of the Department of Methabolic Care and Gerontology, Teaching Hospital in Hradec Kralove. Basic, already developed and validated method was High Performance Liquid Chromatography (HPLC) on reversed-phase column Gemini 5 μm C18 110Ǻ, 150 x 3 mm, Phenomenex (Torrance, USA). Phosphate buffer with concentration of 15 mmol/l, pH 6,4 and flow rate 0,8 ml/min was used as a mobile phase. Diode array detector was used for detection of creatinine (235 nm ) , kynurenine (230 nm) and uric acid (290 nm) Neopterin was monitored using fluorescent detection at 353 nm excitation and 438 nm emission wavelength. All chromatographic systems were compared on the basis of the SST- System Suitability Test and were evaluated by the separation efficiency, peak resolution, asymmetry and other parameters. Columns with the best parameters were selected for further optimization and validation. The resulting new modern HPLC method should be suitable for clinical use.
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Position of Bavaria in the German Economy and Cooperation Possibilities with the Czech Republic.
Prokopová, Veronika ; Zamykalová, Miroslava (advisor) ; Gullová, Soňa (referee)
Diploma thesis deals with position of Bavaria in German economy and cooperation possibilities with the Czech Republic. The main goal of this thesis is to outline the position of Bavaria in German economy and to analyse business environment of Bavaria. Another goal is to find out, what are main negatives of cooperation between the CR and Bavaria and how to improve them. In order to support these findings, a questionnaire was conducted. Also, this thesis suggests possible areas where cooperation could be broadened. This paper presents information about Germany and Bavaria, business environment of Bavaria and possibilities of cooperation.
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The value addition to fresh oranges as a means of reducing post harvest loses and to improve the incomes of the orange farmers in Ghana.
Prokopová, Veronika
The objective of the Bachelors thesis is to analyse value addition to fresh oranges as a means of reducing post-harvest loses and to improve the incomes of the orange farmers in Ghana by reducing waste of raw product and creating possible jobs for local people. The main work deals with health, agriculture, social and educational development in selected areas that are mainly engaged in farming and harvesting oranges. At the end of my thesis I will propose a simple value addition processing for orange farmers and other small scale processor which they can sell to the government for use in the school feed program.
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