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Voltammetric Determination of Simvastatin at a Carbon Paste Electrode and a Silver Solid Amalgam Electrode
Hrochová, Zuzana ; Fischer, Jan (advisor) ; Dejmková, Hana (referee)
This master thesis is focused on determination simvastain by cyclic voltammetry (CV), DC voltammetry (DCV), and differential pulse voltammetry (DPV) at a carbon paste electrode and a silver solid amalgam electrode. The optimum conditions for determination of simvastatin were found and under these conditions, concentration dependences were measured and the limits of detection (LOD) and limits of quantification (LOQ) were calculated for each method. The optimum conditions for determination simvastatin were BR buffer pH 3,0 for DPV and pH 5,5 for DCV and methanolu (20%). For both DCV and DPV the linear concentration dependences were obtained in contentration ranger from 1 to 100 μmol·l-1 with LOD 0. 36 μmol·l-1 and LOQ 1. 2 μmol·l-1 for DCV and LOD 0. 32 μmol·l-1 and LOQ. 1. 09 μmol·l-1 for DPV. Optimal conditions were used for determination simvstatin in drug Simvax 20, Simvacard 20 and Simgal 10. For the determination of simvastatinu in biological fluids a mercury meniscus modified silver solid amalgam electrode was used. Conditions were the same like for measurement conditions for DPV at CPE, but they were modified for reduction region. For ten times diluted urine LOD was 1.83 μmol·l-1 and LOQ 6. 10 μmol·l-1 . For urine without dilution LOD was 0. 65 μmol·l-1 and LOQ 2. 15 μmol·l-1 . The stability...
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Validation of bioanalytical methods - EU and USA legislations (review)
Hrochová, Zuzana ; Matysová, Ludmila (advisor) ; Kujovská Krčmová, Lenka (referee)
This thesis was aimed of the legal requirements and the comparison of specific requirements for validation of bioanalytical methods resulting from the requirement administration Food and Drug Administration (FDA) in the US and European medicines Agency (EMA) in the European Union. It was found that both agencies based on a different legal basis. FDA is part of the federal system. Publishes its own laws and the regulations which have binding legal force . In contrast, the EMA is a non governmental agency and only serves as an advisory comitte of the European Union. Both agencies have issued their instruction - Guids. The FDA issued a Guidance for Industry - Bioanalytical Method Validation and EMA issued Guidaline on bioanalytical method validation. Both manuals are only as recommendations and They haven't legal lability. Both manuals contain similar informations. Both agencies differ from the arrangement of the chapters and an emphasis on individual issues, but the basic parameters that are discussed and Theky are the same selectivity, carry over, LLOQ and petting of the calibration curve, accuracy and precision of the method, system suitability, and cross validation. There were no fundamental differences between the two instructions, they only differ from the criteria that recognize the individual...
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Voltammetric Determination of Simvastatin at a Carbon Paste Electrode and a Silver Solid Amalgam Electrode
Hrochová, Zuzana ; Fischer, Jan (advisor) ; Dejmková, Hana (referee)
This master thesis is focused on determination simvastain by cyclic voltammetry (CV), DC voltammetry (DCV), and differential pulse voltammetry (DPV) at a carbon paste electrode and a silver solid amalgam electrode. The optimum conditions for determination of simvastatin were found and under these conditions, concentration dependences were measured and the limits of detection (LOD) and limits of quantification (LOQ) were calculated for each method. The optimum conditions for determination simvastatin were BR buffer pH 3,0 for DPV and pH 5,5 for DCV and methanolu (20%). For both DCV and DPV the linear concentration dependences were obtained in contentration ranger from 1 to 100 μmol·l-1 with LOD 0. 36 μmol·l-1 and LOQ 1. 2 μmol·l-1 for DCV and LOD 0. 32 μmol·l-1 and LOQ. 1. 09 μmol·l-1 for DPV. Optimal conditions were used for determination simvstatin in drug Simvax 20, Simvacard 20 and Simgal 10. For the determination of simvastatinu in biological fluids a mercury meniscus modified silver solid amalgam electrode was used. Conditions were the same like for measurement conditions for DPV at CPE, but they were modified for reduction region. For ten times diluted urine LOD was 1.83 μmol·l-1 and LOQ 6. 10 μmol·l-1 . For urine without dilution LOD was 0. 65 μmol·l-1 and LOQ 2. 15 μmol·l-1 . The stability...
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Novel Voltammetric DNA Biosensor for the Detection of the DNA Damage Caused by Oncological Drugs
Hrochová, Zuzana ; Vyskočil, Vlastimil (advisor) ; Zima, Jiří (referee)
This Bachelor Thesis is focused on the optimization of conditions for the use of a simple voltammetric DNA biosensor, based on a large-surface carbon film electrode (ls-CFE), for the detection of the DNA damage caused by oncological drugs. ls-CFE was used for its advantageous properties, such as its fast preparation, a simple mechanical renewal of the electrode surface, a good reproducibility of results, a simple chemical modification, and, last but not least, low preparation costs. A content of ethanol in the solution, in which the biosensor was incubated together with the damaging agent, was the main optimization parameter investigated in this Thesis. The contents of ethanol in the range from 0 to 50 % (v/v) of ethanol in 0.1 mol/l phosphate buffer of pH 7.15 (PBS) were tested. After the incubation of the biosensor, the measurements were performed using cyclic voltammetry (CV) in the presence of the redox indicator [Fe(CN)6]4-/3- in PBS. The optimum content of ethanol was found as 5 %. Afterwards, the DNA biosensor, which preparation was optimized in a previous Bachelor Thesis, was used for the detection of the DNA damage caused by model cytostatic agent - Ellipticine (5,11-dimethyl-6H-pyrido[4,3-b]carbazol). Using the CV technique and the redox indicator [Fe(CN)6]4-/3- , the response of the biosensor to...
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