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Study on possible utilization of metal oxide based stationary phases for the analysis of polar compounds
Slaninová, Jitka ; Kovaříková, Petra (advisor) ; Mokrý, Milan (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Student: Jitka Slaninová Supervisor: Doc. PharmDr. Petra Kovaříková, Ph.D. Title of diploma thesis: Study on possible utilization of metal oxide based stationary phases for the analysis of polar compounds This diploma thesis is aimed at investigation of the ability of zirconia (column ZirChrom®-PHASE, 150 x 4,6 mm; 5 µm) and titania (column Sachtopore®-NP, 150 x 4,6 mm; 5 µm) based stationary phases for analyses under the conditions of hydrophilic interaction chromatography (HILIC). Influence of mobile phase strength, mobile phase pH and buffer strength on the separation of biologically important polar compounds: adenosine monophosphate (AMP), adenosine diphosphate (ADP), adenosine triphosphate (ATP), cyclic adenosine monophosphate (cAMP), creatine (C) and phosphocreatine (CP) was tested. All analyses were performed using isocratic elution. The mobile phases consisted of an organic part - acetonitrile (ACN) and an aqueous part - buffer (sodium phosphate dibasic - Na2HPO4 with sodium fluoride - NaF) in different ratios. The mobile phase flow rate was at 1 mL/min. The detection of analytes was performed by using a photodiode array detector at wavelengths of 215 nm for C and CP...
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Analytical evaluation of selected drugs using HPLC
Mingas, Panagiotis - Dimitrios ; Kovaříková, Petra (advisor) ; Kučera, Radim (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Kralove Department of Pharmaceutical Chemistry and Drug Control Candidate: Panagiotis Mingas Supervisor: Assoc. Prof., PharmDr. Petra Kovaříková, Ph.D. Title of diploma thesis: Analytical evaluation of selected drugs using HPLC In the developing pharmaceutical industry, HPLC is one of the most frequently used separation methods. Beyond drug discovery and development, HPLC has many other applications in various fields of pharmaceutical analysis. Aroylhydrazone iron chelators are potential drug candidates which both in vitro and in vivo pharmacodynamics studies demonstrated promising antioxidant and cardioprotective properties. Salicylaldehyde isonicotinoyl hydrazone (SIH) is the compound of the aroylhydrazone class that was the most thoroughly studied. The results that were obtained showed that its hydrazone bond makes SIH susceptible to hydrolysis in biological materials, decreasing its biological half- life. Various novel derivatives were developed in the direction of improving the stability of SIH in biological materials. The main aim of the research was firstly, to develop suitable chromatographic methods for the analysis of 8 novel iron chelators of the aroylhydrazone class (H21, H22, H23, H24, H25, H26, H32, H54) and secondly, utilize...
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Analysis of oxidative and free radical induced DNA damage
Váňová, Nela ; Kučera, Radim (advisor) ; Kovaříková, Petra (referee)
Analysis of oxidative and free radical induced DNA damage Diploma thesis Nela Váňová Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Free radicals and reactive oxygen species (ROS) are highly reactive molecules capable of modifications of biomolecules, including DNA. 5',8-cyclopurine-2'deoxynucleosides represent a group of DNA lesions characterized by concomitant damage to both sugar and base moieties of the same purine nucleoside that are together with 8-oxo-2'-deoxypurines among the major lesions formed by attack of free radicals (e.g. hydroxyl radical). Quantification of oxidative and free radical induced DNA lesions as biomarkers of oxidative stress has a high importance in study of their role in human health and disease. For quantification of these DNA lesions in gamma irradiated samples, high performance liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) will be utilized. Before injection into the LC/MS/MS, irradiated samples, treated by enzymatic digestion in order to gain free nucleosides, have to be desalted and DNA lesions have to be separated from undamaged nucleosides. A new HPLC/UV method was developed for separation of (5'R)-5',8-cyclo-2'- deoxyadenosine;...
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HPLC analysis of daunorubicin and its metabolite
Adamíková, Klára ; Kovaříková, Petra (advisor) ; Kučera, Radim (referee)
At present, the High Performance Liquid Chromatography (HPLC) is still the dominant separation method, it is widely used in all areas of drug analysis. HPLC provides qualitative and quantitative evaluation of the separated components. This thesis describes the development of the method for HPLC analysis of daunorubicin (DAU) and its metabolite daunorubicinol (DAU-OL) in a rabbit plasma. The analyzed compounds were first separated from interfering substances in plasma due the deproteinization. For a precipitation was chosen methanol (600 μl) with the highest extraction efficiency. The separation of DAU and its metabolite was reached by Zorbax SB-Aq column (3,5 µm, 150 x 4,6 mm; Agilent Technologies, USA), the mobile phase consisted of a mixture of water with phosphate butter and phosphoric acid (pH 2,4) and acetonitril (74:26, v/v). The analysis ran at isocratic mode during 15 minutes. The mobile phase was delivered at a rate of 1,1 ml/min, an aliquot of 40 μl was injected into the chromatographic system. The fluorescence detector was operated at an excitation wavelength of 480 nm and an emission wavelength of 560 nm. Doxorubicin was used as an internal standard. The methodology was validated with respect to linearity (4 - 83 ng/ml for DAU, 25 - 500 ng/ml for DAU-OL), precision and accuracy. The...
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Y Soft Campus - the semi-abandoned shooting range area in Brno-Pisarky
Kovaříková, Petra ; Hýl, Petr (referee) ; Kratochvíl, Jan (advisor)
I have designed the campus and the headquarters of the company Y-Soft. It is situated in area Brno-Pisárky. The whole site is 104 900 m large. We have done a research of the needs of employees and management. I have integrated he results into my project. The designed campus includes Y-Soft’s office buildings, a sport gymnasium, a fitness center, a park, residential buildings and kindergarten. The headquarters of Y-Soft is designed as a compact building for easier communication between departments. I tried to create large building with the human scale and nice spaces to work and relax. I have chosen a skeleton construction made from monolithic concrete. The reinforced concrete frame will be insulated by mineral wool. The facade will be created from the part of windows with colored frame and part covered by Thermowood.
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Bioanalytical evaluation of novel drug candidates from the group of thiosemicarbazones II.
Kollárová, Zuzana ; Kovaříková, Petra (advisor) ; Sochor, Jaroslav (referee)
9 ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Candidate: Zuzana Kollárová Supervisor: PharmDr. Petra Kovaříková Title of diploma thesis: Bioanalytical evaluation of new potential agents derived from thiosemicarbazone II. Nowadays, malignant neoplasms (cancer) are listed as the 4th most frequent cause of death. Therefore, its successful therapy and proper drug treatment attract scientific attention all over the world. New potent anticancer agent, 2-benzoylpiridine-4-ethyl-3- thiosemicarbazone (Bp4eT), developed by Prof. D. Richardson (University of Sydney, Australia) was identified as a leading compound of the 2-benzoylpiridine thiosemicarbazone series. These compounds, known as iron chelators, were originally developed to treat metabolic iron overload but recent investigations revealed their antiproliferative properties and highly selective mechanism of action in the therapy of malignant neoplasms. In this study optimal conditions for HPLC-MS analysis of Bp4eT and its degradation products were developed. Also, di(2-pyridilketone)-4-phenyl-3-thiosemikarbazone (Dp4pT) was chosen as an internal standard. Previous studies, during which Bp4eT was incubated with rat and human liver microsomal fractions, confirmed oxidative way...
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Bioanalytical evaluation of novel drug candidates from the group of thiosemicarbazones I.
Šesták, Vít ; Kovaříková, Petra (advisor) ; Kučera, Radim (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of pharmaceutical chemistry and drug analysis Candidate: Vít Šesták Supervisor: PharmDr. Petra Kovaříková, PhD. Title of the Master's Thesis: Bioanalytical assessment of new prospective drugs derived from thiosemicarbazone I. Abstract Development of novel anti-cancer drugs which would boost current therapeutic regimes and/or overcome the resistance against common chemotherapy ranks among the most progressive fields of modern drug discovery. Selective targeting of intracellular iron (Fe) has been recognized as a new strategy for anti-cancer drug development. Biocompatible Fe chelators structurally derived from thiosemicarbazone belong to the most intensively studied anti-cancer agents. Apart from Triapine, currently in phase II of a clinical trial, the systematic investigation led to development of a new generation of iron chelators - di-2-pyridylketone thiosemicarbazones (DpTs). Based on the outcomes of numerous in vitro and in vivo experiments and di-2-pyridylketone-4,4-dimethyl-3-thiosemicarbazone (Dp44mT) was selected as the lead compound for further development. The aim of this study was to develop an LC-MS method for the analysis of Dp44mT and its main phase I metabolites in biological materials (plasma, urine and faeces)....
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Study of retention behavior of polar analytes on novel types of the stationary phases
Buková, Jana ; Kovaříková, Petra (advisor) ; Pilařová, Pavla (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Candidate: Jana Buková Supervisor: PharmDr. Petra Kovaříková, Ph.D. Title of diploma thesis: Study of retention behavior of polar analytes on novel types of the stationary phases. The retention behavior of selected polar analytes (sulphanilic acid, p-hydroxybenzoic acid, p-aminobenzoic acid, 3,4-diaminobenzoic acid, 3-nitrophenol, 3-aminophenol) on two different types of stationary phases in HILIC mode was studied in this diploma thesis. The following stationary phases were used in these experiments: column packed with pure silica Atlantis HILIC Silica, which is widely used for separation in HILIC mode and zirconia column with filling ZirChrom ® - PHASE packing, which is a possible alternative to silica based columns for analysis in HILIC mode, especially due to their high thermal and chemical stability in the whole pH range. The influence of the mobile phase strength on the retention of the selected analytes was tested on both columns. The influence of buffer strength, mobile phase pH and temperature was tested only on zirconia column (due to its higher stability). The chromatographic conditions used in this work were selected according to results of initial experiments.
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Determination of Selected Active Substance in the Preparation VIII
Shkodra, Blerina ; Kastner, Petr (advisor) ; Kovaříková, Petra (referee)
Objective: The purpose of this work was to convert conventional HPLC method to the conditions of UFLC for determination of related substances of Quetiapine Fumarate tablet; new method should need less organic solvents, shorten the analysis time and be validated to guarantee its suitability. Methods: The new chromatographic conditions were assigned using an online method transfer calculator to convert HPLC parameters to UFLC parameters. The suitability of the new method was validated by system suitability test, linearity, quantification limit, detection limit, selectivity, precision and accuracy. Results: The method was shown to be suitable as the tailing factor of the peaks was kept within the limit range of 0.8 - 1.5 and the resolution of each of the peak areas was in all cases greater than 2. In linearity testing, the correlation coefficient obtained from the procedure was 0.999, which showed that the two variables, the peak areas and the range of five different concentrations of the sample highly correlate with each other. The quantification limit obtained from the method was 0.03%, while the detection limit was 0.01%. The selectivity of the method was confirmed as the peak areas of two placebo solutions were shown not to interfere with the peak areas of quetiapine and the impurities dibenzo and...
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Development of an analytical method for evaluation of prodrug- boronyl salicylaldehyde isonicotinoyl hydrazone in a biological material
Bureš, Jan ; Kovaříková, Petra (advisor) ; Sochor, Jaroslav (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Candidate: Jan Bureš Supervisor: PharmDr. Petra Kovaříková, Ph.D. Title of diploma thesis: Development of an analytical method for evaluation of pro-drug - boronyl salicylaldehyde isonicotinoyl hydrazone in biological material High performance liquid chromatography is nowadays one of the most frequently used and progressive analytical method in pharmaceutical analysis. It can be used for both qualitative and quantitative analysis of complicated samples also in a biological matrix. Iron is a vital element for all organisms. It's essential for a proper function of variety of crucial enzymes. On the other hand it's excess in the body is harmful and it is connected with progress of various pathological conditions. Hence iron chelation therapy is an effective tool to treat and protect living cells against injurious effect of iron. Iron chelators were originally used to treat iron overload disease. However, thanks to their interesting antioxidative and antiproliferative properties, their therapeutic potential has been significantly widened. These compounds are currently investigated as promising drugs for the treatment of Parkinson's and Alzheimer's diseases, cancer, malaria,...
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