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Application of modern separation techniques in the analysis of biological material
Červinková, Barbora ; Solich, Petr (advisor) ; Mikuš, Peter (referee) ; Štěrbová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Department of Analytical Chemistry Candidate Mgr. Barbora Červinková Supervisor prof. RNDr. Petr Solich, CSc. Title of Doctoral Thesis Application of modern separation techniques in the analysis of biological material The presented dissertation is focused on development, optimization and validation of new extraction procedures and chromatographic methods for determination of vitamins A and E, inflammation biomarkers, DNA and RNA oxidative stress products in biological matrices. In the theoretical part of this work selected analytes and possibility of their determination in biological materials are described. Moreover, modern trends in liquid chromatography and bioanalytical validation guidelines are included. In this part, special attention is focused on sample handling, transport and pre- treatment. The second part is focused on published methods and divided into two sections. The first section comments published developed analytical methods. As first method, determination of lipophilic vitamins in human serum by miniaturized liquid-liquid extraction with subsequent separation on fluorinated stationary phase using UHPLC system with fluorescent detection in 4 min is discussed. Thereafter, determination of creatinine, DNA and...
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LC-HRMS analysis of selected antihypertensive drugs in biological material for compliance assessment
Tláskalová, Anna ; Kučera, Radim (advisor) ; Štěrbová, Petra (referee)
5 ABSTRACT Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Anna Tláskalová Supervisor - specialist: Mgr. Martin Mžik Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Title: LC-HRMS analysis of selected antihypertensive drugs in biological material for compliance assessment About 40 % of Czech population between the ages of 25 and 64 suffer from arterial hypertension. Although an array of effective antihypertensives is available nowadays, optimal blood pressure during treatment is reached by mere 30 % of the patients. This is mainly due to the patients' poor adherence to the treatment, who use their medicaments incorrectly or not at all. The adherence of a patient to the treatment, as well as pharmacokinetics, represent the most significant source of variability of the answer to the treatment and notably influences its outcome. The monitoring of plasmatic levels of antihypertensives is one of the methods of modern medicine which enables both an effective supervision of the incorrectly compensated patients, and the adjustment of dosage scheme. This diploma thesis focuses on development and optimisation of extraction procedures for selected antihypertensives (amiloride, amlodipine, betaxolol, bisoprolol, carvedilol, celiprolol,...
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The Internal Standard Method for Determination of Ethanol by Head-space Gas Chromatography
Kratochvílová, Eva ; Kučera, Radim (advisor) ; Štěrbová, Petra (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Candidade: Mgr. Eva Kratochvílová Consultant: PharmDr. Radim Kučera, Ph.D. Title of thesis: The internal standard method for determination of ethanol by head-space gas chromatography The theoretical part of this rigorous thesis deals with a literature review focused on the conditions used to determine ethanol by HS-GC. The following settings of HS-GC system were tested in the experimental part of this thesis: optimal time of heating of the sample, temperature and the influence of the matrix on the assay. The method of internal and external standard and the method of standard addition were tested for quantification of ethanol in pharmaceutical products. The method of internal standard was chosen as the optimal one. The following parametres were tested in the validation of the method: linearity, precision, intermediate precision, accuracy, LOD, LOQ and selectivity. The content of ethanol in pharmaceutical product was determined by the validated method.
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Otologics, ophalmologics and dental care in dog and cat
Štěrbová, Petra ; Lamka, Jiří (advisor) ; Ducháček, Lubomír (referee)
This diploma thesis deals with common diseases and difficulties of dogs and cats concerning ears, eyes and teeth with an emphasis on treatment and preventive measures. There is an etiopathogenesis, a symptomatology, a diagnostics and a treatment worked out for each disease. The thesis also describes dosage forms and their routes of administrations. Each chapter includes a list of recorded veterinary pharmaceuticals as well as a list of authorized veterinary pharmaceuticals.
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Analytical and bioanalytical evaluation of novel potential drugs from the group of iron chelators
Bureš, Jan ; Štěrbová, Petra (advisor) ; Kujovská Krčmová, Lenka (referee) ; Hroch, Miloš (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Mgr. Jan Bureš Supervisor: Doc. PharmDr. Petra Štěrbová, Ph.D. Title of Thesis: Analytical and bioanalytical evaluation of novel potential drugs from the group of iron chelators High performance liquid chromatography (HPLC) gained its unprecedented position among bioanalytical techniques due to its effectivity and versatility. If tuned properly, it can separate complex mixtures and can be used for qualitative and quantitative analysis simultaneously, usually within a short time. Reliability of the HPLC methods is proven by their validation performed according to accepted guidelines. Iron chelation is considered to be an effective concept of treatment of various pathologies - from iron overload disease to cancer or neurodegenerative disorders. To develop new chelators for these indications it is important to investigate their structure- activity relationship, the fate of the drug in organism and its relationship to biological effects and toxicity etc. Modern analytical methods are essential tools for these studies. The first part of this work presents validated LC-UV and LC-MS methods for assay of aroylhydrazone iron chelators and related pro-drugs in biological...
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Non-silica based materials in drug analysis III.
Jiřičková, Nina ; Kučera, Radim (advisor) ; Štěrbová, Petra (referee)
Non-silica based materials in the drug analysis III. Diploma thesis Nina Brejchová
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Analysis Sample preparation prior to an essay plays a key role in the drug analysis. Example of a widely used sample preparation technique is a Solid Phase Extraction. This method is characterised by high selectivity, which is given by the type of used sorbent. The aim of this thesis is to study the application of a new sorbent in Solid phase extraction which is zirconium oxide modified by graphene. Graphene is an allotropic modification of carbon possessing a specific adsorptive properties. Therefore it can be used besides other areas in chemical analysis. This thesis focuses on the testing on adsorptive capability of this sorbent using a mixture of model basic drugs. The attention was also paid to pH of solution used for conditioning of the sorbent and type of eluents. After the extraction, the samples were analysed by high- performance liquid chromatography, using a gradient elution. The most appropriate extraction environment was aqueous environment. Extraction recoveries obtained with the prepared sorbent were compared to those obtained with silica gel. The highest recoveries were obtained by extraction of...
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Analytical and bioanalytical evaluation of novel potential drugs from the group of iron chelators
Bureš, Jan ; Štěrbová, Petra (advisor) ; Kujovská Krčmová, Lenka (referee) ; Hroch, Miloš (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Mgr. Jan Bureš Supervisor: Doc. PharmDr. Petra Štěrbová, Ph.D. Title of Thesis: Analytical and bioanalytical evaluation of novel potential drugs from the group of iron chelators High performance liquid chromatography (HPLC) gained its unprecedented position among bioanalytical techniques due to its effectivity and versatility. If tuned properly, it can separate complex mixtures and can be used for qualitative and quantitative analysis simultaneously, usually within a short time. Reliability of the HPLC methods is proven by their validation performed according to accepted guidelines. Iron chelation is considered to be an effective concept of treatment of various pathologies - from iron overload disease to cancer or neurodegenerative disorders. To develop new chelators for these indications it is important to investigate their structure- activity relationship, the fate of the drug in organism and its relationship to biological effects and toxicity etc. Modern analytical methods are essential tools for these studies. The first part of this work presents validated LC-UV and LC-MS methods for assay of aroylhydrazone iron chelators and related pro-drugs in biological...
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Application of modern separation techniques in the analysis of biological material
Červinková, Barbora ; Solich, Petr (advisor) ; Mikuš, Peter (referee) ; Štěrbová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Department of Analytical Chemistry Candidate Mgr. Barbora Červinková Supervisor prof. RNDr. Petr Solich, CSc. Title of Doctoral Thesis Application of modern separation techniques in the analysis of biological material The presented dissertation is focused on development, optimization and validation of new extraction procedures and chromatographic methods for determination of vitamins A and E, inflammation biomarkers, DNA and RNA oxidative stress products in biological matrices. In the theoretical part of this work selected analytes and possibility of their determination in biological materials are described. Moreover, modern trends in liquid chromatography and bioanalytical validation guidelines are included. In this part, special attention is focused on sample handling, transport and pre- treatment. The second part is focused on published methods and divided into two sections. The first section comments published developed analytical methods. As first method, determination of lipophilic vitamins in human serum by miniaturized liquid-liquid extraction with subsequent separation on fluorinated stationary phase using UHPLC system with fluorescent detection in 4 min is discussed. Thereafter, determination of creatinine, DNA and...
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Study of retention properties of polymeric stationary phases
Komůrková, Lenka ; Kučera, Radim (advisor) ; Štěrbová, Petra (referee)
Study of retention properties of polymeric stationary phases Diploma thesis Lenka Komůrková Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Analysis The thesis was focused on the comparison of retention properties of polymer stationary phases based on acrylate and styrene with reversed phases based on zirconium dioxide and commonly used silica gel C18. Cyclosporine A (CsA) and his accompanying impurities B (CsB), C (CsC) and D (CsD) were used model analytes. Measurements were carried out in the analytical scale to obtain basic data for the employment of polymeric phases as the appropriate sorbents in preparative chromatography with regard to their high volume commercial availability. The effect of organic solvent was tested on the separation and separation efficiency. The main attention was focus on the methanolic mobile phases (MF) in the case of polymeric phases due to the economic reasons. All impurities were separated on column silica gel C18 (MF = methanol 80 %) and polymethacrylate C18 (MF = methanol 71 %). Impurity CsB was not separated on column ethylstyrene-divinile copolymers (DVB) (MF = methanol 90 %) and moreover the co-elution of CsD with CsA occured. Peak tailing was observed in a methanolic mobile phase on the...
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Study on possibility of utilization of alternative sorbents for sample preparation
Kresová, Jana ; Štěrbová, Petra (advisor) ; Kučera, Radim (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Candidate Mgr. Jana Kresová Consultant Doc. PharmDr. Petra Kovaříková, Ph.D. Title of Thesis Study on possibility of utilization of alternative sorbents for sample preparation Sample preparation belongs to basic operations during biological sample analysis. Overall success of analytical determination depends on its accomplishment. Nowadays, Solid Phase Extraction (SPE) is positively a dominant technique in the field of biological materials sample preparation. Graphene is one of the new forms of carbonaceous materials. Due to its extraordinary properties (large surface area, structure rich in π-electrons, good thermal and chemical stability) it became a suitable adsorbent in analytic sample preparation. The aim of this work was to evaluate extraction recoveries of several sorbents: ZirChrom® -SHAX (SHAX, polyethyleneimine-coated zirconia), ZirChrom® -SAX (SAX, polyethyleneimine-coated zirconia - less cross-linked than SHAX), ZirChrom® -PEZ (PEZ, ethylendiamine-N,N,N',N'-tetramethylenphosphoric-acid-coated zirconia), aminosilica (AS) both unmodified and graphene-oxide modified followed by thermal reduction. The extraction efficiency was tested using the model mixture containing...
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