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Vliv vybraných látek ovlivňujících centrální nervový systém na kostní metabolismus
Fekete, Soňa ; Živný, Pavel (advisor) ; Horák, Pavel (referee) ; Nobilis, Milan (referee)
The increase in life expectancy of the world population is associated with challenges regarding health issues. For instance, osteoporosis is a medical condition mostly observed in elderly people, in which the quality and quantity of the bone are severely affected. Not only for women but also for men, osteoporosis is recognized as an important public health issue. Osteoporosis is a systemic skeletal disorder and is a result of loss of skeletal mass. Osteoporosis is characterized by low bone mass, microarchitectural deterioration of bone tissue and an increase in bone fragility and susceptibility to fracture. Osteoporotic fractures are a significant cause of morbidity and mortality. The longterm use of drugs such as antiepileptics and antidepressants could affect the onset of osteoporosis. The aim of the study was to evaluate the effect of orchidectomy, the effect of newer antiepileptic (levetiracetam, lacosamide, topiramat, lamotrigine) and antidepressive drugs (mirtazapine, venlafaxine and trazodone) on bone metabolism in healthy male Wistar rats. The first specific aim was to determine the effect of orchidectomy on bone metabolism in rats. We found that after 12 weeks post-orchidectomy there was a negative effect on bone metabolism in rats. These results established these animals as suitable models for...
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Evaluation of biologically active substances using HPLC IV.
Seidlová, Martina ; Pilařová, Pavla (advisor) ; Nobilis, Milan (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department Pharmaceutical Chemistry and Drug Kontrol Name of Degrese Paper: Evaluating of biologically active substances by HPLC IV. Candidate: Martina Seidlová Tutor: PharmDr. Pavla Pilařová, Ph.D. In this diploma work the study of dilation behaviour of medications was provided under extreme conditions for a column being able to measure pH ranged 1-12 on column Kinetex EVO C18 (150 x 3 mm). For higher pH the following medications were studied: quetiapine, and its two metabolites - norquetiapine and 7-hydroxyquetiapine, bromhexine, codeine, zolpidem and its admixture, fluoxetine and aciclovir. The best results were reached with quetiapine with its metabolites, therefore they were chosen for the following tests and possibility of separation from plasma. As a mobile phase an acidic buffer was used at pH10 using ratio 70:30 to organic phase which was made of acetonitrile and methanol (55:5). For lower pH the following medications were studied: salicylic acid, acetylsalicylic acid, aciclovir, bromhexine and fluoxetine. The best results were reached with salicylic acid and acetylsalicylic acid, therefore they were chosen for the following testing and the possibility of separation in the medicament. As a mobile phase a phosphate buffer...
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The Development of SPE Extraction Procedure for Isolation of Pravastatin and its Metabolite from Biological Matrix
Rabatinová, Martina ; Nováková, Lucie (advisor) ; Nobilis, Milan (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Martina Rabatinová Supervisor: Doc. PharmDr. Lucie Nováková, Ph.D. Title of Rigorous Thesis: The development of SPE extraction procedure for isolation of pravastatin and its metabolite from biological matrix Statins are a class of drugs used to lower cholesterol levels. Statins have an inhibitory activity against 3-hydroxy-3-methylglutaryl coenzyme A reductase. The main purpose of this work was to develop and validate a quantitative method for sensitive analysis of pravastatin and pravastatin lactone. The analytes were extracted from rat urine by a solid phase extraction method. In this method urine samples were prepared on C18 Discovery DSC-18. Washing and elution agents consisted of a mixture of acetonitrile and 0.01 M ammonium acetate buffer, pH 4.5. Number of extraction steps were optimized to eliminate most of matrix effects. The method involved use of ultra high performance liquid chromatography tandem mass spectrometry. Chromatography was carried out on a BEH C18 1.7 µm, 2.1×100 mm column. A mobile phase consisted of a mixture of acetonitrile and 0.5 mM ammonium acetate solution, pH 4.0. Linear gradient conditions were used. Total run time of this anal- ysis was 6 minutes...
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Development and Validation of HPLC Methods for Transport and Metabolic Studies
Hroch, Miloš ; Nobilis, Milan (advisor) ; Šatínský, Dalibor (referee) ; Šišpera, Luděk (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Candidate Ing. Miloš Hroch Consultant Doc. PharmDr. Milan Nobilis, CSc. Title of Thesis Development and validation of HPLC methods for transport and metabolic studies The presented PhD thesis deals with the development and validation of chromatographic methods in bioanalysis of drugs and their metabolites. The work provides a theoretical introduction to the field of bioanalysis, using methods of liquid chromatography. Attention was paid to recent trends in the area, particularly with respect to the new types of stationary phases. In the experimental part of the thesis, reader is introduced to a development and validation of several bio-analytical methods using high performance liquid chromatography. The first method concerns with a determination of 5-fluorouracil and its metabolites by HPLC with gradient elution and UV/VIS detection on a column Luna PFP (2). Discussed are mainly specifics for the assay of this group of polar and poorly detectable compounds. The developed and validated method is being used for TDM of patients with rectal cancer. Another discussed method deals with a determination of methotrexate polyglutamates in erythrocytes using isocratic elution on a hybrid...
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Analyses of Statins in Biological Samples
Vlčková, Hana ; Solich, Petr (advisor) ; Nobilis, Milan (referee) ; Šimek, Petr (referee)
5 ABSTRACT Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Mgr. Hana Vlčková Supervisor: Prof. RNDr. Petr Solich, CSc. Supervisor specialist: Assoc. Prof. PharmDr. Lucie Nováková, Ph.D. Title of Doctoral Thesis: Analysis of statins in biological samples The doctoral thesis deals with analysis of statins in biological samples. Statins are the most widely used drugs for the treatment of Familiar hypercholesterolemia. Due to very low concentration of statins (ng/ml) in human plasma a choice of suitable analytical methods is limited. Liquid chromatography with mass spectrometry detection is the most suitable technique due to high selectivity and sensitivity. Sample preparation step has to be used before chromatographic determination of analytes in complex matrices, such as biological samples. Primarily theoretical part of doctoral thesis summarizes individual pharmacological properties and effects of statins. Another part is devoted to theoretical knowledge of UHPLC-MS/MS. The widest section deals with description of the individual sample preparation techniques. The practical part contends with development of UHPLC-MS/MS methods for the determination of statins in biological samples and selection and development of suitable sample preparation...
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Quantitative Measurement of Drugs in Biological Samples using Chromatography Methods.
Maláková, Jana ; Nobilis, Milan (advisor) ; Polášek, Miroslav (referee) ; Chládek, Jaroslav (referee)
Charles University in Prague, Faculty of Pharmacy in Hradci Králové Department of Pharmaceutical Chemistry and Drug Control Candidate: PharmDr. Jana Maláková Supervisor: Doc. PharmDr. Milan Nobilis, PhD., Associate Professor of Bioorganic Chemistry. Title of Doctoral Thesis: Quantitative measurement of drugs in biological samples using chromatography methods. The thesis deals with the application of instrumental analytical methods to the quantitative measurement of drugs and their metabolites in biological samples using liquid and gas chromatography. Various procedures of sample preparation were evaluated during optimization of analytical assays. Precipitation of proteins with acetonitrile was employed (analysis of cefuroxime), liquid-liquid extraction (analysis of topiramate, 8-methoxypsoralen) and samples were also purified using a mixed-mode solid-phase extraction (reversed-phase and ion-exchange principle) during the analysis of galantamine and its metabolites. The unbound cephuroxime fraction in the plasma and protein binding were determined using an ultrafiltration method. Chromatography conditions - gas or liquid chromatography- depended on physico-chemical characteristics of a drug. Due to the thermal decomposition of topiramate, derivatization was used to form a more stable methyl...
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Analysis of Biologically Active Compounds by Capillary Electrophoresis
Petrů, Klára ; Polášek, Miroslav (advisor) ; Nobilis, Milan (referee) ; Urbánek, Marek (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate Mgr. Klára Petrů Supervisor Doc. RNDr. Miroslav Polášek, CSc. Title of Doctoral Thesis Analysis of biologically active compounds by capillary electrophoresis The thesis presented is dealing with the analysis of quaternary ammonium compounds of pharmaceutical importance by capillary electrophoresis (CE). Basic principles of CE and respective means of detection are discussed in the theoretical part. Special attention is paid to capacitively coupled contactless conductivity detection C4D and to the CE coupling with mass spectrometry (CE-MS). The chemical and pharmacological properties of the compounds of interest: carbethopendecinium bromide, pancuronium bromide (PM), vecuronium bromide (VM) and rocuronium bromide (RM) are characterized. The experimental work is focused on the development and validation of new analytical CE methods for the assay of such compounds. in pharmaceuticals. The C4D detection was applied because of poor UV adsorption capability of all compounds studied. I. The first part deals with the CE analysis of septonex, an antiseptic drug with the character of surfactant. The background electrolyte was 30 mM MES of pH 7 containing 12.5 mg/ml of 2-hydroxypropyl-β-...
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Modern Trends in Sample Preparation of Biological Material for Analysis of Selected Biologically Active Compounds
Sadílek, Petr ; Solich, Petr (advisor) ; Nobilis, Milan (referee) ; Barták, Petr (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Petr Sadílek, M.Sc. Supervisor: Prof. RNDr. Petr Solich, CSc. Co-supervisor: Doc. RNDr. Dalibor Šatínský, Ph.D. Title of Doctoral Thesis: Modern Trends in Sample Preparation of Biological Material for Analysis of Selected Biologically Active Compounds This doctoral thesis deals with the determination of selected drugs, their metabolites and other biologically active substances in biological materials. This area of instrumental analysis turns out to be very attractive recently. The reason for this is the need to monitor and optimise treatment and the goal is to achieve the most effective therapy with minimum side effects of drugs for every individual; there are also toxicological reasons. The thesis concentrates on the entire analytical phase of laboratory screening, i.e. not just on development and validation of chromatographical methods of analysis, but also (and especially) on preparation of biological samples prior to the actual analysis. This phase is crucial for analytes assessment, since it determines not only correct results of the laboratory examination, but also influences the lifespan of laboratory equipment, especially chromatographic columns. These days mostly traditional...
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Transdermal Permeation Enhancers: Study of Structure-activity Relationships.
Novotný, Michal ; Vávrová, Kateřina (advisor) ; Nobilis, Milan (referee) ; Rabišková, Miloslava (referee)
Permeation enhancers are compounds that promote drug flux into systemic circulation or adjacent tissues. This doctoral thesis follows our previous results and aims at studying two different groups of potential permeation enhancers: dicarboxylic acid derivatives and analogues of Transkarbam 12. The main purpose of my work was to study the structure- activity relationships in these permeation enhancers and to contribute to the elucidatation of their mechanism of action. The first part of my work was focused on derivatives of dicarboxylic acids. Series of enhancers based on maleic, fumaric, succinic, tartaric and meso-tartaric acids were prepared. The enhancing activity of these compounds was investigated in vitro in Franz diffusion cells using theophylline as a model drug and porcine skin. The single-chain amphiphiles were markedly more effective than the double-chain ones. The most active enhancer, monododecyl maleate, that is a cis- derivative, was a more potent enhancer than its respective trans- isomer. Furthermore, the activity of similar succinates, i.e. compounds having a single bond, strongly depended on the donor vehicle. No difference between diastereoisomeric tartaric and meso-tartaric acid derivatives was found. The second part of my work dealt with Transkarbam 12, an ammonium carbamate...
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Study of chiral aspects of drug metabolism using instrumental analytic methods
Holmanová, Veronika ; Nobilis, Milan (advisor) ; Kovaříková, Petra (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Candidate: Bc. Veronika Holmanová Supervisor: Doc. PharmDr. Milan Nobilis, CSc. Title of diploma thesis: Study of chiral aspects of drug metabolism using instrumental analytic methods For the evaluation of stereospecificity of rat and human cytosolic carbonyl reductases and activity of microsomal oxidases involved in nabumetone biotransformation, a new chiral high-performance liquid chromatographic method was developed. The prepared LLE-HPLC-PDA method enabled extraction, separation and ultraviolet detection of prochiral nabumetone and its five phase I metabolites including enantiomers of two chiral biotransformation products of carbonyl reduction. Methyl ester of naproxen served as an internal standard. Diethyl ether was used for liquid-liquid extraction (LLE) of biomatrices. Chiralcel OD-R 250 mm×4.6 mm column with a mobile phase methanol-1M NaClO4/HClO4 aqueous solution pH=3 (75:25, v/v) were employed in isocratic sufficient separation of nine analytes. The whole analysis lasted 60 minutes at the flow rate of 0.5 ml/min. The column effluent was monitored using a photodiode-array detector (scan or single wavelength at λ=265 nm). The results of in vitro nabumetone...
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