|
|
The use of supercritical fluid chromatography in pharmaceutical analysis
Babičová, Barbora ; Khalikova, Maria (advisor) ; Matysová, Ludmila (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Barbora Babičová Supervisor: Mgr. Maria Khalikova, CSc. Consultant: doc. PharmDr. Lucie Nováková, Ph.D. Title of the Diploma Thesis: The use of supercritical fluid chromatography in pharmaceutical analysis Keywords: enzalutamide, ticagrelor, supercritical fluid chromatography, method validation Enzalutamide is a new drug for the treatment of metastatic form of prostate cancer. Ticagrelor is a drug with antiplatelet activity. The optimization of UHPSFC/UV methods for analysis of both active substances enzalutamide, and ticagrelor, and their respective impurities is described in diploma thesis. In the case of enzalutamide, there were 4 impurities analysed using following conditions: Viridis BEH 2-EP column (3.0 x 100 mm, 1.7 µm) with a column temperature of 40 řC. The mobile phase used was methanol with 10 mM ammonium formate and gradient elution using 0-40 % of organic mobile phase modifier over 3 minutes with a flow rate of 1.5 ml/min. The UV detection was proceed at 270 nm. The total analysis time was 4.5 minutes. Separation of ticagrelor and its 2 impurities was made using Acquity UPLC HSS C18 SB column (3.0 x 100 mm, 1.8 µm). The mobile phase was composed of methanol and acetonitrile (1:1,...
|
|
|
Implementation of new analytical system into a routine biochemical laboratory
Berdych, Martin ; Khalikova, Maria (advisor) ; Hyšpler, Radomír (referee)
IN ENGLISH Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of analytical chemistry Candidate: Bc. Martin Berdych Supervisor: Mgr. Maria Khalikova, Ph.D. Consultant: RNDr. Jiří Plíšek, Ph.D. Title of the diploma thesis: Implementation of new analytical system into a routine biochemical laboratory The diploma thesis deals with the procedures in the case of the exchange of a biochemical analyzer under conditions of common laboratory clinical practice in a biochemical laboratory. The experimental part is focused on the comparison of the COBAS Integra 800 and COBAS 6000 analytical systems and the assessment of the degree of conformity in the measurement of biochemical methods using statistical tools (differential analysis according to Bland-Altman and Passing-Bablok regression analysis). With the help of determining the partial uncertainties of the measurement results, it evaluates the verification of individual methods by measuring control and reference materials on the COBAS 6000 analyzer. Comparison of eight methods - alanine aminotransferase, antistreptolysin-O, aspartate aminotransferase, C-reactive protein, HDL cholesterol, magnesium, creatine kinase and LDL cholesterol - a certain degree of difference appeared to have been assessed. Consequently some of the...
|
|
|
Implementation of new analytical system into a routine biochemical laboratory
Berdych, Martin ; Khalikova, Maria (advisor) ; Hyšpler, Radomír (referee)
IN ENGLISH Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of analytical chemistry Candidate: Bc. Martin Berdych Supervisor: Mgr. Maria Khalikova, Ph.D. Consultant: RNDr. Jiří Plíšek, Ph.D. Title of the diploma thesis: Implementation of new analytical system into a routine biochemical laboratory The diploma thesis deals with the procedures in the case of the exchange of a biochemical analyzer under conditions of common laboratory clinical practice in a biochemical laboratory. The experimental part is focused on the comparison of the COBAS Integra 800 and COBAS 6000 analytical systems and the assessment of the degree of conformity in the measurement of biochemical methods using statistical tools (differential analysis according to Bland-Altman and Passing-Bablok regression analysis). With the help of determining the partial uncertainties of the measurement results, it evaluates the verification of individual methods by measuring control and reference materials on the COBAS 6000 analyzer. Comparison of eight methods - alanine aminotransferase, antistreptolysin-O, aspartate aminotransferase, C-reactive protein, HDL cholesterol, magnesium, creatine kinase and LDL cholesterol - a certain degree of difference appeared to have been assessed. Consequently some of the...
|
|
|
Determination of illegal compounds in cosmetics by modern chromatographic methods
Novotná, Aneta ; Khalikova, Maria (advisor) ; Matysová, Ludmila (referee)
Charles University, Faculty of Pharmacy Department of Analytical Chemistry Candidate: Aneta Novotná Supervisor: Dr. Maria Albertovna Khalikova, Ph.D. Consultant: doc. RNDr. Dalibor Šatinský, Ph.D. Title of Graduation Thesis: Determination of illegal substances in cosmetics using modern chromatographic methods This diploma thesis deals with the development of ultra - fast liquid chromatographic method, which will best detect the preservatives used in cosmetics (parabens). The development of the method involves the selection of the best stationary phase and the mobile phase. In the next phase, selecting the appropriate temperature, gradient, flow rate, and before applying real cosmetic samples, validation was required to show that the method is sufficiently accurate and repeatable for this research. This development of the method was performed on the UltiMate ™ 3000 HPLC and UHPLC System, 9 stationary phases from 4 manufacturers (Fortis, Ascentis, Kinetex, Onyx), properties and contents were evaluated. The most suitable stationary phase that separated all 9 substances was Kinetex Biphenyl 100A, 2.6 μm 150 x 4.6 mm. Due to the solubility and properties of the parabens, two mobile phases and one combination thereof (ACN, MEOH) were tested. The acetonitrile-containing mobile phase better separated the...
|
|
|
An influence of individual modes of TOF analyzer on linearity and sensitivity of UHPLC-HRMS methods
Halad, Jakub ; Vlčková, Hana (advisor) ; Khalikova, Maria (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Jakub Halad Supervisor: RNDr. Hana Vlčková, Ph.D. Title of thesis: An influence of individual modes of TOF analyzer on linearity and sensitivity of UHPLC-HRMS methods This diploma thesis deals with the comparison of four modes (sensitive mode, resolution mode, high resolution mode, enhanced resolution mode) of the hybrid analyser Q- TOF connected with UHPLC system. Totally, ten analytes were selected for this study. It was a group of 5 statins and their interconversion products. UHPLC method was borrowed from previous developed multistatin method however, ESI conditions were optimized. Developed method was used to measure standard calibration curves and matrix calibration curves in four analyser modes (sensitive, resolution, high resolution and enhanced mode). Spiked lyophilized serum treated by the protein precipitation was used for matrix calibration curve. Finally, the individual analyser modes were compared due to the linearity and sensitivity The effect of individual analyser modes on the correlation coefficient was not observed. On the other hand, analyser modes influenced the method sensitivity. In positive mode, lower LOQ values were obtained for the standard calibration curves...
|
|
|
Development and validation of method for the determination of agomelatin in tablets using UHPSFC and UHPLC
Plachká, Kateřina ; Nováková, Lucie (advisor) ; Khalikova, Maria (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Kateřina Plachká Supervisor: doc. PharmDr. Lucie Nováková, Ph.D. Title of Diploma Thesis: Development and validation of methods for the determination of agomelatine using UHPSFC and UHPLC Agomelatine is one of the newest antidepressants. Due to a different mechanism of action it offers a completely new approach in the treatment of depressive disorders. Two chromatographic methods for determination of agomelatine and its impurities were developed. The separations on UHPSFC system were accomplished using stationary phase based on BEH 2-EP and gradient elution using CO2 and MeOH containing 20mM ammonium formate and 5 % of water. The gradient was run from 5 - 30 % of organic modifier in 3 min. The UHPLC separations were performed on stationary phase BEH Shield RP18. The mixture of acetonitrile and methanol in ratio 1:1 and 10mM ammonium acetate buffer pH 9.5 were used as mobile phase. Both developed methods were properly validated in terms of linearity, sensitivity (LOD, LOQ), accuracy and precision according to ICH guidelines. The UHPSFC method was linear in the range 0.25-70 μg/ml for all analytes with method accuracy ≥ 97.4 % and ≥ 100.2 % and method precision RSD ≤ 2.4 and ≤ 0.8 for...
|
|
|
Determination of illegal compounds in sport supplements by modern chromatographic methods
Žaludová, Michaela ; Khalikova, Maria (advisor) ; Nováková, Lucie (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Michaela Žaludová Supervisor: Dr. Maria Albertovna Khalikova, Ph.D. Consultant: doc. RNDr. Dalibor Šatinský, Ph.D. Title of Diploma Thesis: Determination of illegal compounds in sport supplements by modern chromatographic methods This diploma thesis concerns on the development of method for the determination of steroid hormones in sport supplements using ultra-high performance supercritical fluid chromatography. The method development and optimization of chromatographic conditions include selection of a suitable stationary phase and a modifier together with a mobile phase additive. Furthermore, the influence of temperature, pressure, flow rates and mobile phase gradient on separation efficiency was examined. In conclusion, this method has been validated and applied on real samples. Experiments were carried out on the device ACQUITY UPC2 with PDA detector. Detection was at wavelength 232 nm. 11 compounds were analysed. The stationary phases screening included 5 different analytical columns ACQUITY UPC2 BEH 1,7 µm, BEH 2-EP 1,7 µm, HSS C18 1,7 µm, CSH Fluoro-Phenyl 1,7 µm and 2-PIC 1,7 µm dimensions 3,0 x 100 mm. As the most suitable was selected column HSS C18 1,7 µm (3,0 x 100 mm)....
|
|
|
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...
|
|
|
Evaluation of the influence of various solvents for column storage in supercritical fluid chromatography
Lidvinová, Kateřina ; Nováková, Lucie (advisor) ; Khalikova, Maria (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Kateřina Lidvinová Supervisor: Assoc. Prof. PharmDr. Lucie Nováková, Ph.D. Title of Diploma Thesis: Evaluation of the influence of various solvents on column storage for supercritical fluid chromatography The diploma thesis deals with the evaluation of the influence of various solvents on column storage for supercritical fluid chromatography. It is focused especially on column aging which is manifested as reduced retention time and the change in selectivity of a stationary phase when stored for a certain period of time. The experiments were performed using the ACQUITY UPC2 SFC system with the PDA detector. The detection took place at wavelength of 225 nm for the mixture of neutral compounds and at wavelength of 230 nm for the mixture of ionisable compounds. The ACQUITY UPC2 BEH 1.7 µm and the ACQUITY UPC2 BEH 2-EP 1.7 µm columns with dimensions of 3.0 x 100 mm (Waters, USA) were used as stationary phases. After the separation, these columns were always flushed with different solvents and stored, namely in CO2, isopropanol, acetonitrile and hexane in which they were stored until the next use. CO2 with methanol as an organic modifier was used as a mobile phase for the mixture of...
|
guest ::
