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Introduction of spectrophotometric determination of urinary citrates and oxalates into routine clinical laboratory testing.
FEJT, Martin
Determination of the level of citrates and oxalates in urine is of great importance in the diagnosis and subsequent monitoring of patients with urolithiasis. Data about urine concentrations of these substances, together with other metabolic tests, help to determine the likelihood of recurrence of urolithiasis. At the same time, they help physicians find appropriate regime measures for patients with the disease. This bachelor thesis deals with the introduction of the determination of citrates and oxalates in the routine operation of a clinical laboratory. The theoretical part first briefly describes the issue of urolithiasis, its distribution, etiopathogenesis, treatment and prophylaxis. Emphasis is placed on the role of citrates and oxalates. Furthermore, individual types of urinary stones and possibilities of their analysis are characterized. The following is an approach to the process of verification of the measuring procedure and associated performance parameters. The related analytical properties of the methods are also listed and described here. Within the description of the implementation process the setting of parameters in the laboratory and hospital information system is briefly explained. In the laboratory of ÚKBH FN Plzeň we chose enzymatic methods and an automatic analyzer Beckman Coulter AU480 to determine the concentration of citrates and oxalates in urine. The theoretical part of the bachelor thesis continues with a description of the spectrophotometry and operation of the instrument used. We tested reagents from foreign manufacturers INstruchemie (Netherlands) and LTA (Italy). The following is an explanation of the principles of the methods from the above manufacturers and briefly lists other possible methods for the determination of these analytes. Reagents from INstruchemie were selected for routine use. As this manufacturer has already validated its methods on Beckman Coulter spectrophotometers, only their verification has taken place at ÚKBH. The practical part of this bachelor thesis deals first with the preparation of reagents for analysis and continues with programming the parameters of individual methods into the analyzer. The main content of this work is a detailed description of the implementation of individual phases of analysis, which are required by the verification process. After successful completion of all measurements and evaluation of their results, appropriate verification protocols were prepared and the methods were put into routine operation of the laboratory.
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Search for an efficient cobalt chelator
Nosková, Karolína ; Mladěnka, Přemysl (advisor) ; Smutný, Tomáš (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Karolína Nosková Supervisor: prof. Přemysl Mladěnka, PharmD., Ph.D. Consultant: Monika Moravcová, MSc. Title of diploma thesis: Search for an efficient cobalt chelator Cobalt, as part of vitamin B12, is an essential trace element for human organism. However, long-term exposure to high doses leads to toxicity. Chronic cobalt intoxication is currently most commonly observed after the implantation of cobalt-containing prostheses, which in some cases do not work properly and cobalt is released from them by mechanical friction or corrosion. That not only leads to a local reaction, but cobalt ions also enter the blood and lymphatic circulation and thus cause a systemic reaction. The intoxication is then manifested by neurological, cardiovascular and endocrine symptoms, which can lead to irreversible changes or even to the death. However, a chelator that would be able to effectively and selectively bind cobalt and thus effectively treat intoxication is not available in clinical practice. The aim of this thesis was a search for an efficient cobalt chelator among known metal chelators. Chelation was measured in vitro by a method based on spectrophotometric detection of unchelated cobalt ions at...
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Search for an efficient cobalt chelator
Nosková, Karolína ; Mladěnka, Přemysl (advisor) ; Smutný, Tomáš (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Karolína Nosková Supervisor: prof. Přemysl Mladěnka, PharmD., Ph.D. Consultant: Monika Moravcová, MSc. Title of diploma thesis: Search for an efficient cobalt chelator Cobalt, as part of vitamin B12, is an essential trace element for human organism. However, long-term exposure to high doses leads to toxicity. Chronic cobalt intoxication is currently most commonly observed after the implantation of cobalt-containing prostheses, which in some cases do not work properly and cobalt is released from them by mechanical friction or corrosion. That not only leads to a local reaction, but cobalt ions also enter the blood and lymphatic circulation and thus cause a systemic reaction. The intoxication is then manifested by neurological, cardiovascular and endocrine symptoms, which can lead to irreversible changes or even to the death. However, a chelator that would be able to effectively and selectively bind cobalt and thus effectively treat intoxication is not available in clinical practice. The aim of this thesis was a search for an efficient cobalt chelator among known metal chelators. Chelation was measured in vitro by a method based on spectrophotometric detection of unchelated cobalt ions at...
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The Flow Injection Analysis of Procaine and Sulfamethoxazole with Spectrophotometric Detection
Baptistová, Adéla ; Nesměrák, Karel (advisor) ; Červený, Václav (referee)
A derivatization reaction, based on a copulation reaction of diazonium salt of an analyte with a reagent (1-aminonaphthalene or NEDA) producing colour, spectrophotometrically detectable product, has been used for determination of two analytes (procain hydrochloride, sulfamethoxazole) containing amino group in the molecule. The conditions of batch spectrophotometric determination were studied. It was found, that the batch determination is applicable only for procain hydrochloride analysis, because sulfamethoxazole gives time-unstable colour product. FIA arrangement was proposed and effects of selected parameters were optimized using two levels factorial design. The calibration dependences were measured for both analysis with both reagents, and subsequently were applied on determination of analytes in medical forms. Key words: diazotization, flow injection analysis, procaine, spectrophotometry, sulfamethoxazole
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Determination of Procaine by Flow Injection and Sequential Injection Analysis with Spectrophotometric Determination
Tomanová, Marie ; Hraníček, Jakub (advisor) ; Rychlovský, Petr (referee)
This diploma thesis is focused to the determination of procaine using flow injection and sequential injection analysis coupled with spectrophotometric detection. This determination is based on the reaction of procaine with a colouring agent, 1,2-naphthoquinone-4-sulfonic acid. An orange coloured product is formed, which is determined spectrophotometrically at the wavelength 484 nm. The high of the absorption signal of the product is directly proportional to the concentration of procaine. The aim of this work was to optimize the parameters of both methods of flow analysis so that the limit concentration of procaine can be as low as possible and at the same time, high sensibility is achieved. The next step was to apply these methods on the determination of procaine in real samples. It was found that in flow injection analysis, the absorbance of procaine hydrochloride obeys Beer's law for concentrations from 2.5 to 120 µg/ml. The linear regression equation of calibration graph was y = 0.0059x - 0.0051, with a linear regression correlation coefficient 0.9993. Limit of detection was 0.72 µg/ml. Effects of standing time (stopped-flow), flow rate, concentration of colouring agent, pH and the volume of the sample loop have been examined and optimized. It was also found that in sequential injection...
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The comparison of methods for determination of ascorbic acid in medical formulations
Kohoutová, Petra ; Nesměrák, Karel (advisor) ; Červený, Václav (referee)
The aim of this thesis is to compare and assess chosen analytical methods for ascorbic acid determination in pharmaceuticals with respect to their correctness, accuracy, time and finance exigencies. Four methods were chosen alkalimetric titration with visual and instrumental indication, iodometry and spectrophotometry. Pills, capsules and injection solution were chosen as samples of pharmaceuticals. The obtained results as to accuracy of the declared value (or range permitted under Pharmacopoeia) agreed by all titration methods, spectrophotometric determination showed overestimated results. All used methods were approximately equally accurate. As time seems to be most optimal acidobasic titration with visual indication, while demanding in terms of time was spectrophotometry. If we consider methods on financial intensity all titration methods were quite equal.
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Physico-Chemical Properties of Drugs
Tomanová, Pavla ; Kubíček, Vladimír (advisor) ; Nováková, Veronika (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of biophysics and physical chemistry Author: Pavla Tomanová Supervisor: Ing. Vladimír Kubíček, CSc. Thesis title: Physico-Chemical properties of drugs This thesis deals with measuring dissociation constants (the pKa value) and the lipophilicity of nitrogenous heterocycle derivatives produced at the Faculty of Pharmacy in Hradec Králové. Two methods were used to determine the pK value: potentiometric titration and spectrophotometry. Potentiometric titration almost invariably produced relatively exact results, however the results of pK value obtained by spectrophotometry were sometimes difficult to replicate. Dissociation constants were compared on the basis of the method applied, and also with the theoretically determined dissociation constant calculated by means of the ChemDraw Ultra 12.0 software. The results obtained by the two experimental methods turned out to be very similar, corresponding to the theoretical pK value. Lipophilicity is another essential property of drugs tested in the preclinical stage. It specifies the aqueous or lipophilic phase affinity of a drug, substantially affecting the drug's ADMET in organism. Lipophilicity was determined on the basis of capacity factor value identified by RP-HPLC measurement using...
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Ability of copper chelators to interact with iron and zinc
Hanuščinová, Lucia ; Mladěnka, Přemysl (advisor) ; Čonka, Patrik (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Lucia Hanuščinová Supervisor: Assoc. Prof. Přemysl Mladěnka, PharmDr., PhD. Title of diploma thesis: Ability of copper chelators to interact with iron and zinc Copper plays in the human organism a role of an element with indispensable significance, whose biological influence and effects depend on its quantity. With elevated concentrations in the human body, copper becomes toxic, resulting in pathological conditions. The most well-known diseases is the Wilson's disease, whose treatment consists of oral administration of chelators, i.e. chemical compounds, which are capable of binding copper ions in various proportions and eliminating them from the organism. Chelation therapy is currently the first choice after confirmation of the diagnosis. Chelation toxicity results from several factors, e.g. inhibition of copper dependent enzymes or low selectivity to metals. And precisely the selectivity of chelators is being discussed in this diploma thesis. An ideal chelator should not interact with any of the other physiological ions, that are necessary for the proper functioning of the organism. Five of the most frequently therapeutically or experimentally used substances /trientine, D-penicillamine,...
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Multicomponent Analysis
Kovářová, Lucie ; Kubíček, Vladimír (advisor) ; Drastík, Martin (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Lucie Kovářová Supervisor: Ing. Vladimír Kubíček, CSc. Title of Diploma Thesis: Multicomponent analysis This diploma thesis deals with the determination of concentrations of individual dyes in mixtures by multicomponent spectrophotometric analysis. The method was tested on selected dyes which were divided into two groups. The first group contains methyl orange, methyl red, orange II and phenol red. The second group consists of methylene blue, bromocresol green and thymol blue. Within each group mixtures were prepared containing combinations of two, three and four dyes in various ratio. Spectra of prepared mixtures were measured by diode array UV/VIS spectrophotometer. Absorbance values were read at wavelengths of absorption maxima of the individual dyes. Using the pseudoinverse matrix, concentration ratios of components were calculated from the absorbance values. The determination in determined and over determined systems was performed. On the basis of the results obtained by the method of analysis it is possible to determine the individual concentration of the multi-component dyes mixtures without prior separation. Keywords: spectrophotometry, multicomponent analysis,...
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Search of effective cobalt chelators - 8-hydroxyquinolines
Pelnářová, Karolína ; Mladěnka, Přemysl (advisor) ; Carazo Fernández, Alejandro (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Karolína Pelnářová Supervisor: Prof. Přemysl Mladěnka, PharmD., Ph.D. Consultant: Václav Tvrdý, MSc.; Marcel Hrubša, MSc. Title of diploma thesis: Searching of effective cobalt chelators - 8-hydroxyquinolines Cobalt is one of the essential trace elements present in the human body. It forms a part of the organometallic complex of vitamin B12, which is essential for many physiological functions. Both overload and lack of cobalt in the body is associated with pathological conditions. Manifestations of deficiency can lead up to pernicious anemia or hypofunction of the thyroid gland. Cobalt intoxication can occur in an industrial environment, such as by cobalt metal dust during heavy metal processing, or by its release due to corrosion from orthopedic prostheses. Systemic toxicity is manifested by a number of endocrine, cardiovascular and neurological symptoms. The main aim of this diploma thesis was to find effective chelators of cobalt from the group of substances derived from 8-hydroxyquinoline. In vitro spectrophotometric measurement was used to determine the degree of chelation. The effect of chelation was also monitored ex vivo in rat erythrocytes. All tested chelators - 8-hydroxyquinoline,...
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