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Development of UHPLC-MS/MS and UHPSFC-MS/MS methods for determination of selected ochratoxins
Kretek, Kristián ; Nováková, Lucie (advisor) ; Chmelařová, Hana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Kristián Kretek Supervisor: prof. PharmDr. Lucie Nováková, Ph.D. Title of the diploma thesis: Development of UHPLC-MS/MS and UHPSFC-MS/MS methods for determination of selected ochratoxins The purpose of this work was development and comparison of UHPLC and UHPSFC analytical methods coupled to tandem mass spectrometry for four common ochratoxins. The experiments were performed using UHPLC system ACQUITY UPLC and UHPSFC system ACQUITY UPC2, mass spectrometer XEVO TQ-XS was the same in both cases. First, optimization of ion source parameters and SRM transitions was done. For UHPLC along with ACN, 3 additives of the water-based component of mobile phase were tested - formic, acetic acid and ammonia, with flow rate 0,4 ml/min, gradient from 2 % to 95 % ACN in 5th minute, temperature 40řC and column Acquity UPLC BEH C18. For UHPSFC 6 organic modifiers along with CO2 were tested: MeOH and MeOH with addition of water, formic acid, ammonia, and ammonium formate, with flowrate 1,5 ml/min, gradient from 2 % to 45 % MeOH in 5th minute, temperature 40řC and BPR pressure 2000 psi. Screening of 13 columns was also conducted. Optimization of MS detection in SFC was done by testing 8 make-up solvents: MeOH, EtOH,...
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Favorit Brno / cycling stadium - bicross track / architectural study - design / stage 2021/22
Nováková, Lucie ; Žalmanová, Petra (referee) ; Šindlar, Jiljí (advisor)
The assignment of the diploma thesis is an architectural study of a cycling stadium - velodrome in the Pisárky district in Brno for TJ Favorit Brno. The existing velodrome is located southwest on the edge of the Brno Exhibition Center on Křížovského Street. The Velodrome no longer meets the current requirements of track cycling, but it is historically valuable and has its genius loci. The new 250 m long wooden bike track (together with other parameters of the International Cycling Union) and its associated sports grounds are "inserted" into the existing velodrome, which is modified and reconstructed according to requirements. The grandstands have been preserved. The built-in building with a new track is then completely roofed, creating an enclosed space. As part of the stadium reconstruction, a new facility was also created for TJ Favorit Brno from a modular container system; These are changing rooms for athletes, administrative areas, and facilities for visiting clubs and athletes and IP areas. The container construction system for sports facilities is very practical, economical, fast, environmentally friendly, variable, and flexible to the current requirements of a constantly evolving sport. The Anteroom and ground floor in the southern part of the existing velodrome is equipped with services for the public: a cafe, souvenir shop, cash register, information center, service, and a bicycle and sports equipment store. The new track with grandstands is covered with ETFE foil anchored to new lattice trusses, which are connected to the existing roof.
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Determination of neopterin and its derivatives by HPLC with fluorescence detection.
Říhová, Lenka ; Nováková, Lucie (advisor) ; Chocholoušová Havlíková, Lucie (referee)
TITLE: Determination of neopterin and its derivates by HPLC with fluorescence detection SUMMARY: Neopterin, a pteridine derivative, is produced in human organism by monocytes or by B - lymphocytes stimulated with interferon γ or interleukine - 2. These active substances (interferon γ, interleukine - 2) are connected with immune system activation. Neopterin measurement in body fluids can therefore serve for monitoring of activation of various components of immune system induced by cytokines. Immune activation occurs for example during many infectious diseases, autoimmune diseases and malignant diseases. Recently the connection between neopterin and 7,8 - dihydroneopterin and intracellular oxidative stress and cell apoptosis was demonstrated in several scientific studies. Values of pteridines probably correlate with certain neurological disesases. Totally reduced form of biopterin serves as cofactor of some enzymes in human organism. In clinical practice the concentration of neopterin, eventually other related substances is expressed as the ratio neopterin / creatinine. This diploma thesis is dealing with the development of an analytical method for the determination of biologically active substances biopterin, 7,8 - dihydroneopterin, 5,6,7,8 - tetrahydroneopterin, neopterin and creatinine by high performance...
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Stanovení residuí fluorochinolonů v půdách metodou LC-FD
Aufartová, Jana ; Solich, Petr (advisor) ; Nováková, Lucie (referee)
7. Abstract Several pharmaceutics used in human and veterinary medicine could be detected in environment. To these compounds belongs Fluoroqinolones. Our work observed 4 fluoroqinolons (ofloxacin, norfloxacin, ciprofloxacin, enrofloxacin) in soils sample. Fluoroquinolones were determinate by LC-FD. For extraction was used mixture of organic compounds (methanol: acetone; 1:1) and 1.0g of EDTA. Sample was 15 minutes shacked, 15 minutes in ultrasonic bath and 15 minutes of centrifugation. After that organic solution was evaporated by gentle stream of nitrogen in water bath. Rest from evaporation was dissolved in mQ-H2O and clean-up was done. After evaporation after clean-up the rest was dissolved in mobil phase. Mobil phase was H3PO4: MeOH: ACN (920: 70: 10) and flow was 1.4mL/min. Limit of quantification (LOQ) is For OFLO 5mg/L, for NOR is 0.083mg/L, for CIP is 0.116mg/L and for ENRO is 0.125mg/L. Mean recoveries ranged between 75% to 121%, for OFLO, NOR, CIPRO and ENRO. At the beginning we tried also acidic type of extraction. Extraction was made with HCL, H2SO4, H3PO4 and HNO3. In all cases we add EDTA because of interferences with other substances in samples. We find out, that no complex is build between flouroquinolones and EDTA. For clean- up from soil samples is better OASIS column HLB 6cc/200mg....
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Matrix effects in LC-MS analysis: occurrence, evaluation, and their elimination
Svoboda, Pavel ; Nováková, Lucie (advisor) ; Švec, František (referee) ; Havliš, Jan (referee)
Abstract Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Pavel Svoboda Supervisor: Assoc. Prof. PharmDr. Lucie Nováková, Ph.D. Advisor: RNDr. Hana Vlčková, Ph.D. Title of Doctoral Thesis: Matrix effects in LC-MS analysis: occurrence, evaluation, and their elimination The dissertation thesis deals with the issue of matrix effects in analysis using liquid chromatography coupled to mass spectrometry (LC-MS). LC-MS represents a well-established, widely used, and powerful tool in the field of qualitative and quantitative analysis of a wide range of different compounds. Despite all the advantages offered by LC-MS such as sensitivity, selectivity, and robustness, it suffers from a major drawback represented by matrix effects. This undesirable phenomenon affects important parameters of LC-MS method such as accuracy, precision, linearity, limit of detection, and limit of quantification. Therefore, an evaluation of these effects should be involved in validation of every single LC-MS method and their source should be eliminated. The assessment is generally performed by qualitative and quantitative approaches such as post-extraction addition, post-column infusion, and comparison of slopes of calibration curves. The matrix effects can be reduced or...
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Testing of Chromatographic Qualities of Alternative Stationary Phases
Havlátová, Michaela ; Šatínský, Dalibor (advisor) ; Nováková, Lucie (referee)
1 Abstract Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Michaela Havlátová Consultant: Doc. RNDr. Dalibor Šatínský, PhD. Diploma Thesis Title: Testing of Chromatographic Qualities of Alternative Stationary Phases Chromatographic qualities of HS F5 column were tested at selected groups of substances. For testing there were chosen substances from the following groups: beta-sympatholytics (acebutolol, bopindolol), neuroleptics (thioridazine, perphenazine), local anesthetics (lidocaine, prilocaine), tetracyclins (oxytetracyclin, minocyclin), methylxanthine derivates (caffeine, theophylline) and aniline. During the analysis the retention of the tested substances was investigated as influenced by the character of the mobile phase. Some of the substances showed U-shaped retention. Three series of analysis were carried out at each tested group. In analysis number 1 acetonitrile - water was used as the mobile phase, in analysis number 2 the mobile phase was acetonitrile - phosphoric acid 0.085%, and in analysis number 3 it was acetonitrile - acetate buffer solution pH 7. In each analysis the acetonitrile percentage in the mobile phase was changed. In the group of the substances tested the U-shaped retention was shown by beta-sympatholytic...
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Characterization of separation systems for determination of enantiomers
Geryk, Radim ; Tesařová, Eva (advisor) ; Ševčík, Juraj (referee) ; Nováková, Lucie (referee)
(EN) The dissertation thesis is focused on the research and characterization of retention and enantiorecognition mechanisms of chiral stationary phases based on derivatized polysaccharides. The separation systems with a variety of modern stationary phases (both achiral and chiral) were characterized in detail to provide a comprehensive view of the interactions participating in the separation process. The study of the retention/separation behavior significantly facilitates the development and the optimization of new enantioselective methods for a wide variety of compounds. The work deals with the comparison of enantioselective performance of polysaccharide-based chiral stationary phases. The objectives are to show the differences of separation behavior among these chiral stationary phases, as they differ by the nature of the polysaccharide backbone (amylose versus cellulose), by binding of chiral polymer to silica support (coated versus immobilized stationary phase) and by the phenyl moiety in the reversed and normal phase HPLC. In both separation modes amylose-based chiral stationary phases exhibited higher enantioselectivity, especially for acidic and bifunctional analytes. Chiral stationary phases based on derivatized cellulose showed higher enantiodiscrimination potential for basic analytes....
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The development of method for the determination of atorvastatin and its impurities in tablets using supercritical fluid chromatography
Juchelka, Jan ; Nováková, Lucie (advisor) ; Khalikova, Maria (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Jan Juchelka Supervisor: doc. PharmDr. Lucie Nováková, Ph.D. Title of Diploma Thesis: The development of method for the determination of atorvastatin and its impurities in tablets using supercritical fluid chromatography This diploma thesis was focused on developement and optimization of a method for qualitative and quantitative evaluation of atorvastatin and its impurities using supercritical fluid chromatography. One of the main goals was to optimize chromatographic conditions, which included the selection of a sufficiently selective stationary phase, appropriate modifier and mobile phase additives. Effect of pressure was also explored. Finally this method was validated. A total of seven analytes were evaluated, lately only six of them. One impurity was impossible to separate without using a chiral stacionary phase since it was an enantiomer of atorvastatin. Measurements were performed on UPC2 Acquity system with PDA detector by Waters. Selected wavelength for detection was 245 nm. Tested stationary phases included columns dedicated for SFC, namely Acquity UPC2 BEH 1,7 µm, BEH 2-EP 1,7 µm, HSS C18 SB 1,8 µm a CSH Fluoro-Phenyl 1,7 µm (all of them 3,0 x 100 mm). Insufficient separation...
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Development of UHPLC-MS/MS method for the determination of statins and their metabolites
Svoboda, Pavel ; Nováková, Lucie (advisor) ; Novák, Ondřej (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Pavel Svoboda Supervisor: Doc. PharmDr. Lucie Nováková, Ph.D. Title of Graduation Thesis: Development of UHPLC-MS/MS method for the determination of statins and their metabolites This graduation thesis deals with development of the method for the determination of statins and their metabolites by ultra-high performance liquid chromatography coupled to tandem mass spectrometry. First, the chromatographic conditions were optimized. Next, the parameters of mass spectrometer were optimized. And finally, repeatability, linearity and sensitivity of the method were assessed. Chromatographic column BEH C18 (50 x 2.1 mm, 1.7 µm) was used for the analysis. The choice of column was performed using UV spectrophotometric detector. The mobile phase of the gradient elution consisted of 0.1 mM ammonium acetate (A) and acetonitrile (B). The initial ratio of mobile phase A:B was set at 70:30. The optimization of the mass spectrometry parameters started with the selection of precursor ions using direct infusion. Thereafter, the ion source parameters of the mass spectrometer were optimized. The ion transitions were optimized after the selection of product ions. The optimization of ion transitions was...
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Optimalization of sample preparation step for UHPLC-MS/MS analysis of atorvastatin, rosuvastatin and their metabolites
Pilařová, Veronika ; Nováková, Lucie (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Veronika Pilařová Supervisor: Doc. PharmDr. Lucie Nováková Ph.D. Title of GraduationThesis: Optimalization of sample preparation step for UHPLC-MS/MS analysis of atorvastatin, rosuvastatin and their metabolites The purpose of this graduation thesis was the optimalization of UHPLC-MS/MS method for the determination of concentrations of atorvastatin, rosuvastatin and their metabolites in biological material, and then development and optimalization of a MEPS (microextraction by packed sorbent) method for sample preparation of biological material and validation of this method. Acquity BEH C18 column (50 x 2.1 mm, 1.7 µm, Waters) was used for the separation of the analytes. Electrospray ionization was performed in both negative and positive ion mode. Triple quadrupole mass analyser was used for detection. Precursor ions and fragment ions were chosen for each statin. Collision energy and cone voltage were optimized for all analytes individually. Quantification of analytes was performed using the SRM (selected reaction monitoring). Protonated molecule [M+H]+ , which was measured in the positive ion mode, was chosen as a precursor ion for rosuvastatin lactone. Ion [M-H]- , which was measured in...
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