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Determination of adrenaline and noradrenaline in plasma by HPLC
Kučerová, Tereza ; Klapková, Eva (advisor) ; Kotaška, Karel (referee)
The target of this bacalary work was to validate the metod for determination of adrenaline and noradrenaline in plasma by high performance liquid chromatography with electrochemical (coulometric) detection. The metod was first necessary to optimize for the coulometric detection because it was used the Bio-Rad kit, which was designed to determine with amperometric detection. The principle of this metod is the chromatographic separation of substances with subsequent oxidation of the determined analyte passes through the analytic cell with coulometric detection. The part of the analytical procedure is the solid phase extraction. The part of this work was the optimization of the metod for the coulometric detector. For the analysis were selected following potentials: channel 1 - 350 mV with range 100 nA, channel 2 - 0 mV with range 50 nA, guard channel 0 mV with range 50 nA. Furthermore, the chromatographic metod was fully validated (accuracy, precision, limit of detection, robustness). In the conclusion was made interlaboratory comparison of results of 42 patients samples with those obtained in the hospital Šternberk.
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Comparison of gel and capillary electrophoresis of proteins
Krbec, Michal ; Franěk, Tomáš (advisor) ; Kotaška, Karel (referee)
Electrophoresis of serum proteins is one of the methods used in laboratory diagnosis of pathological conditions, but also for research purposes. This is a one-step method, which involves isolation of individual proteins from a mixture (mostly blood proteins) in the electric field in the presence of a suitable buffer and a suitable carrier based on their different mobility, dependent on their surface charge and molecular weight. Originally the carrieer used to be a paper, later cellulose acetate membrane and now it is almost exclusively agarose (purified agar with low endoosmosis) or polyacrylamide (PAA). My task in this work was to analyze a serie of samples by gel electrophoresis and subsequently analyze the same serie by capillary electrophoresis. Results obtained by both methods I worked numerically, compared with each other and statistically evaluated. The output of my work is a comparison of gel and capillary electrophoresis, evaluation of the advantages, disadvantages and especially the differences between these two methods.
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The posibility of inorganic sulphate determination in patients with severe metabolic disease
Plasová, Martina ; Kotaška, Karel (advisor) ; Zadina, Jiří (referee)
Inorganic Sulphate play important role in various metabolic pathways. The aim of the work was to assess the posibility of inorganic sulphate determination in patients with severe kidney diseses undergoing hemodialysis. Serum sulphate levels were determined in 68 patients (31 men, mean age 67, range 31 - 86 and 37 women mean age 71, range 44 - 86) before and after hemodialysis. Two turbidimetric methods Quantichrom TM Sulfate Assay Kit and BGR gelatine method were used for determination. Both methods were performed separately in group of 34 patients. Results were as follows: Mean serum sulphate levels in patients before hemodialysis are significantly elevated than the levels after hemodialysis (p<0.0001, non parametric t-test, alpha = 0.05). BGR gelatine method show less significant differences in sulphate levels before and after hemodialysis than the Quantichrome Assay method. (p = 0.037 vs. p < 0.001, non parametric t-test, alpha = 0.05 ). This fact could be explained by the significant role of interferences in BGR gelatine method. We confirm the relevance of both metod in inorganic sulphate determination. BGR gelatine method is more affected by interference, than the QuantiChrome Assay method. Powered by TCPDF (www.tcpdf.org)
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Investigations of uratic concrements in urolithiasis
Kultová, Lenka ; Zadina, Jiří (advisor) ; Kotaška, Karel (referee)
Urolithiasis is wide - spread disease, which can lead to serious complications as far as acute renal failure. It is important to analyze the composition of calculus for accurate diagnosis and effective prevention. I deal with analysis of urinary stones by infrared spectroscopy and polarization microscopy in this work. In the first part of the work I intent on the methodical aspects of assesment. The set of ten samples, which included rife components of calculi, was evaluated by two software programs - Kidney Stone Analysis and Omnic. Programme Kidney Stone Analysis gives just one end - result. Programme Omnic indicates ten possible results of the constitution of the sample, which best matches with spectra from libraries. The evaluative person has a possibility to compare these results and decide for the end - result, according to how quantitative data in the evaluation repeat. The result of component's content around 10 % can not be regard as totally reliable. The polarization microscope enables to check and specify the results of the analysis. To use a combination of both methods claimed to be very profitable. In the second part of the work, patients' urinary stones were analyzed, in the way how samples are evaluated in laboratory running. Patient group was formed by 20 persons (17 men and 3 women) in the...
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Laboratory determination of bone markers in the course of osteoporosis
Moučková, Štěpánka ; Kotaška, Karel (referee) ; Dršata, Jaroslav (advisor)
1 Abstrakt Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Štěpánka Moučková Supervisor: ing. Marta Pechová Title of bachelor thesis: Laboratory determination of bone markers in osteoporosis The aim of this study is to compare preliminary values of bone formation and resorption markers in newly diagnosed osteoporotic patients with values after their treatment. We compared the serum levels of N-MID osteocalcin and aminoterminal propeptide of procollagen type I (PINP) as markers for bone formation, and serum carboxyterminal telopeptide (β-CTx) as a marker of bone resorption. Data were collected from 66 patients - 60 women (47 postmenopausal and 13 premenopausal) and 6 men. Patients were divided into 4 groups according to age and applied treatment. Analysis of individual markers was performed using Roche Cobas e411 immunochemical analyzer. Simultaneously were measured serum levels of 25-OH vitamin D3, kalcium (TCa) and parathyrin. Results show that patients treated with bisphosphonates have a significant change in markers of bone turnover, while in patients without proven osteoporosis the levels of bone markers remain unchanged after the supplementation of vitamin D and calcium. Values of vitamin D without supplementation are often below the...
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