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LC-MS monitoring of selected antimicrobial compounds in waters
Krupka, Dominik ; Kučera, Radim (advisor) ; Kastner, Petr (referee)
5 Abstract Charles University Phaculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Dominik Krupka Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Consultant: Ing. Martin Ferenčík Title of diploma thesis: LC-MS monitoring of selected antimicrobial compounds in waters The monitoring of residues of human and veterinary antibiotics in surface water is gaining more and more importance due to the increasing resistance of bacteria. This work deals with the modificate of an HPLC-MS/MS method to obtain pilot data on the occurrence of antibiotics in surfaces water. The method is focused on the analysis of 30 antibiotics from the groups of β-lactams, macrolides, lincosamides, amphenicols, fluoroquinolones, tetracyclines and sulfonamides. The method was performed on a UHPLC instrument Agilent 1200 Infinity series with MS Agilent model 6495 triple Quad. Separation of antibiotics was performed on reverse phase using gradient elution and tandem mass spectrometry. An electrospray in both positive and negative polarity was used to ionize the analytes. The method was used for the pilot analysis of surface water samples in the Czech Republic. Samples from 15 rivers and streams were analysed. The most frequently found antibiotics were sulfomethoxazole and...
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Guest house in Lipnice nad Sázavou
Černý, Martin ; Kučera, Radim (referee) ; Bantová, Sylva (advisor)
The subject of my master´s thesis is project of Guesthouse in Lipnice nad Sázavou. It consists three parts: documentation for building permission, technical equipment and facility management. The building is L-shaped and it is designed as a two-storey building with partial basement. In basement is a technical facilities, engine rooms and storage rooms. On the ground floor there is an entrance to the building with reception. There is also fitness and restaurant for guests. The first floor is part for guests, there are twelve rooms, two playrooms, common room and cleaning room. The individual floors are accessible by stairs or lift. The building is set in a slightly sloping terrain. There are three entrances, for guests, employees and separate entrance to restaurant. Another entrance is to the bike shed. The foundation structures are made of plain concrete foundation strips. The main construction system is brick and the load-bearing structures and some non-bearing structures are from fired clay blocks. Other non-bearing structures are made of plasterboard. The horizontal load-bearing floor structure consists of a reinforced concrete slab. The construction of the gable roof is made of hammered wooden trusses. The land around the building is used for guests' leisure time. It is possible to spend time on the outdoor terrace of the restaurant, playground and outdoor pool. There are also paved sidewalks and parking for guests and employees. It is also planned for the disabled. The main part of project was done using ArchiCAD software.
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The uset of HPLC in the field of chiral separations VII.
Štilcová, Kristýna ; Kučera, Radim (advisor) ; Pilařová, Pavla (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department: Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Student: Kristýna Štilcová Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Consultant: Mgr. Ondřej Horáček Title of Thesis: The employment of HPLC in the field of chiral separations VII. Boron clusters are inorganic, synthetically prepared, three-dimensional, cage-like structures. Boron cluster compounds which are made only of boron and hydrogen atoms are completely symmetrical. Their symmetry can be disrupted by endo- or exoskeletal substitution resulting in chiral compounds. Carboranes, subgroup of the boron cluster compounds, contain at least one carbon atom in their structure and usually include exoskeletal substitution. Therefore, the metallacarboranes containing cobalt bis(dicarbollides) and 7,8-dicarba-nido-undecaborates that have been studied can be found among these chiral structures. Thanks to the specific properties such as high lipophilicity, metabolic stability and delocalized negative charge, the studied compounds can be used as isosteric substitution of phenyl ring in pharmacophores. Due to the growing interest in cobalt bis(dicarbollides) and 7,8-dicarba-nido-undecaborates and the importance of chirality in pharmacy, suitable conditions for chiral...
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Optimization of derivatization reaction conditions for GC-MS analysis of selected compounds
Jan, Tomáš ; Kučera, Radim (advisor) ; Váňová, Nela (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Candidate: Tomáš Jan Title of Thesis: Optimization of derivatization reaction conditions for GC-MS analysis of selected compounds Quinolinic acid is an endogenous molecule, downstream product of the kynurenine pathway, through which the amino acid L-tryptophane is metabolised. Quinolinic acid acts as an agonist at the NMDA receptor. Overactivity of this receptor leads to the cell death of neurons. Current research shows the connection between a high level of quinolinic acid and a higher risk of neurodegenerative diseases development and behavioural disorders. The level can be increased by the excessive intake of tryptophane (e.g. food supplements declaring calming effect and mental health support) or ongoing inflammation in the organism. The current work deals with the development of an analytical method for the determination of quinolinic acid in various biological matrices using GC/MS and complements methods analysing other metabolites of tryptophan. Derivatization by alkyl chloroformate was met with success. The selectivity was compared on column RTX-5MS containing non-polar stationary phase and polar column SLB-IL59 using ion...
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Development of LC-MS method for determination of selected endocrine disruptors
Chvalkovská, Eliška ; Kučera, Radim (advisor) ; Štěrbová, Petra (referee)
Charles University Phaculty od Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Eliška Chvalkovská Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Consultant: Mgr. Martin Novák Title of thesis: Development of LC-MS method for determination of selected endocrine disruptors The thesis is focused on the development and optimization of LC-MS method for the determination of twelve endocrine disruptors in amniotic fluid of pregnant women in the third trimestr of pregnancy. There is a suspision that premature births are connected with an increased incidence of endocrine disruptors in amniotic fluid. The analytes were extracted by liquid-liqud extraction with 4-methylpentan-2- on. The analytical method is based on using reverse phase high-performance liquid chromatography with Hypersil C4 GOLD column (2.1 × 100 mm, particles 1.9 µm). For analysis the gradient elution with the mobile phase A (0.5% (v/v) formic acid aqueous solution) and B (0.5% (v/v) formic acid solution in acetonitrile) at a flow rate of 0.5 ml/min, injection 5 µl and column oven temperature 35 řC was used. The high- resolution mass spectrometer Orbitrap Q Exative Plus with atmospheric pressure chemical ionization in positive mode was used to identify and determine the response of...
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LC-MS/MS study of phase one in vitro biotransformation of potential drugs against Alzheimer's disease
Kuřátková, Aneta ; Kučera, Radim (advisor) ; Váňová, Nela (referee)
No treatment that would completely stop the progression of Alzheimer's disease has not been found yet. Recently used tacrine showed good results in the treatment of Alzheimer's disease, however long-term use led to chronic hepatotoxicity due to its metabolites. This master thesis deals with the compound 7-phenoxytacrine, one of the promising tacrine derivatives, which is one of the candidates for potential use in the therapy of Alzheimer's disease. Due to the formation of hepatotoxic metabolites of tacrine after the biotransformation in human liver, it appears necessary to identify the emerging metabolites of 7-phenoxytacrine molecule. Within this master's thesis in vitro biotransformation study of 7-phenoxytacrine using human liver microsomes was performed. High performance liquid chromatography with tandem mass spectrometry was used to determine the parent substance and the seventeen 7-phenoxytacrine metabolites. The analytical method showed the formation of six monohydroxylated and eleven dehydroxylated metabolites of 7-phenoxytacrine. Thus, we concluded that hydroxylation is the major metabolic reaction after in vitro microsomal biotransformation. In addition to the identification of metabolites, a quantification and microsomal stability study, including the determination of the amount of...
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Validation of chromatographic methods in pharmaceutical analysis
Kegkeroglou, Iokasti ; Kučera, Radim (advisor) ; Kastner, Petr (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Student: Iokasti Kegkeroglou Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Title of Thesis: Validation of chromatographic methods in pharmaceutical analysis Validation is an integral part of every analytical method. Its aim is to demonstrate that the method is suitable for the intended use. This work provides an overview and comparison of documents related to the validation of bioanalytical methods. The work includes guidelines that are currently valid in Europe, the USA or have general validity. At first, attention is paid to the history and developments in this area. Subsequently, the parameters that need to be tested in the validation study are described and are divided according to the regulatory authorities. The following chapter compares the latest versions of the guidelines regarding the validation of separation bioanalytical methods issued by the EMA, FDA, and ICH. Although the individual methodologies are similar in many aspects, there are still differences among them. Hopefully, the differences will be eliminated in the framework of harmonization and only one methodological guideline could be used worldwide. Finally, it follows a detailed statistical evaluation of...
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HPLC evaluation of tyrosine and its metabolites
Michailellis, Panagiotis ; Kastner, Petr (advisor) ; Kučera, Radim (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Author: Panagiotis Michailellis Supervisor: PharmDr. Petr Kastner, PhD. Title of Diploma Thesis: HPLC evaluation of tyrosine and its metabolites Tyrosine is an important precursor of catecholamines, which are dopamine, norepinephrine (noradrenaline), and epinephrine (adrenaline). These are neurotransmitters and hormones, crucial for every living organism. Therefore, their identification and evaluation in biological material would aim to understand their function and behavior more accurately. This Diploma Thesis is a review on how to evaluate catecholamines and their metabolites in biological matter with analytical methods used in Pharmaceutical Analysis, especially with HPLC. First chapters present theoretical knowledge about metabolism of tyrosine and its metabolites, how a sample from an organism should be treated in order to be examined, information about HPLC, CE, GC apparatuses and the main detectors used in analysis of these compounds. In literature review, tables are presented, concerning different sample preparation methods, HPLC details and characteristics, and finally a sum of all the analytical methods studied for this Diploma Thesis. All the articles and...
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Development of LC-MS method for determination of selected endocrine disruptors
Chvalkovská, Eliška ; Kučera, Radim (advisor) ; Štěrbová, Petra (referee)
Charles University Phaculty od Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Eliška Chvalkovská Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Consultant: Mgr. Martin Novák Title of thesis: Development of LC-MS method for determination of selected endocrine disruptors The thesis is focused on the development and optimization of LC-MS method for the determination of twelve endocrine disruptors in amniotic fluid of pregnant women in the third trimestr of pregnancy. There is a suspision that premature births are connected with an increased incidence of endocrine disruptors in amniotic fluid. The analytes were extracted by liquid-liqud extraction with 4-methylpentan-2- on. The analytical method is based on using reverse phase high-performance liquid chromatography with Hypersil C4 GOLD column (2.1 × 100 mm, particles 1.9 µm). For analysis the gradient elution with the mobile phase A (0.5% (v/v) formic acid aqueous solution) and B (0.5% (v/v) formic acid solution in acetonitrile) at a flow rate of 0.5 ml/min, injection 5 µl and column oven temperature 35 řC was used. The high- resolution mass spectrometer Orbitrap Q Exative Plus with atmospheric pressure chemical ionization in positive mode was used to identify and determine the response of...
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Optimization of derivatization reaction conditions for GC-MS analysis of selected compounds
Jan, Tomáš ; Kučera, Radim (advisor) ; Váňová, Nela (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Candidate: Tomáš Jan Title of Thesis: Optimization of derivatization reaction conditions for GC-MS analysis of selected compounds Quinolinic acid is an endogenous molecule, downstream product of the kynurenine pathway, through which the amino acid L-tryptophane is metabolised. Quinolinic acid acts as an agonist at the NMDA receptor. Overactivity of this receptor leads to the cell death of neurons. Current research shows the connection between a high level of quinolinic acid and a higher risk of neurodegenerative diseases development and behavioural disorders. The level can be increased by the excessive intake of tryptophane (e.g. food supplements declaring calming effect and mental health support) or ongoing inflammation in the organism. The current work deals with the development of an analytical method for the determination of quinolinic acid in various biological matrices using GC/MS and complements methods analysing other metabolites of tryptophan. Derivatization by alkyl chloroformate was met with success. The selectivity was compared on column RTX-5MS containing non-polar stationary phase and polar column SLB-IL59 using ion...
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