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Development and Validation of New HPLC Methods for Determination of Biologically Active Compounds in Clinical Practice
Krčmová, Lenka ; Solich, Petr (advisor) ; Sochor, Jaroslav (referee) ; Malíř, František (referee)
Abstroct Charles University inPrague, Foculty of Phormocy in Hrodec Králové Deportment of Ánolytícol Chamistry Condídote: M9r. Lenko Krčmovó Supervisor: prof. RNDr. Petr Solich, CSc. Title oÍDoctorol Thesis: Developmant ond volidotion of new HPLC methods for determinotion of bíolo9icolly octive compounds in clinicol proctísa The presented dissertotion thesis is daolin9 with developmenŤ ond volidotion of HPLC methods for clinical research ond clinicol proctice in Reseorch loborotory of Daportment of Metobolic Care and Gerontology (GMC) in University Hospitol in Hrodec Krolove (UHHK). For this purpose modern technologies deoling with lorge sequences of biologicol moteriol, speed seporotion, smoll volum es of organic solvalt ond somple were used. Naw HPLC method for deŤerminotion of 13-cis ond oll-trons retinoic acid, retinol, alpho- ond gommo-tocopherol using intarnol colibrotion for potients suffers from ocute promyelocytic leukemio treated with retinoic ocid wos daveloped ond volidoted. Solid phose extroction (SPE) ond monoliths were selected for somple praporotion ond seporotion of the target compounds. New method for determinotion of neopterin, creatinine, kynuranine ond tryptophon in humon serum using two monolithic column os stotionory phose wos developed ond volidoted. Thís method wos developed...
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Potenciometrické stanovení gentamycinu ve farmaceutických přípravcích
Kočí, Martina ; Pospíšilová, Marie (advisor) ; Polášek, Miroslav (referee)
Gentamicin is an aminoglycoside antibiotic, and can treat many types of bacterial infections, particularly Gram-negative infection. It is one of the heat-stable antibiotics that remain active after autoclaving, which make it particularly useful in the preparation in the certain microbiological growth media. Traditionally, pharmaceutical analysis of these compounds relies heavily on chromatography, yet also frequently requiring the use of reagent-based detection principles like spectrophotometric, chemiluminescent detection in flow-injection analysis , capillary electrophoresis, a novel LC/ELSD, thin-layer chromatography, HPLC.A simple and sensitive potentiometric procedure for the determination of gentamicin is presented, after development of selective electrodes based on PVC membranes using different type of cyclodextrins and its derivatives, or Evans Blue complex with gentamicin as ionophers. Several mediator solvents and lipophilic ionic species were also used in order to optimize the characteristics of the membranes. The evaluation of the general characteristics for all the units prepared revealed an analytical linear range of about 2x10-4; 5x10-3 mol L-1, slopes of about 50 mV/dec, good reproducibility and low response times. The evaluation of Potentiometric selectivity coefficients will permit to...
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Application of chromatographic methods for the separation of metabolites of cholesterol
Tetřevová, Lucie ; Nováková, Lucie (advisor) ; Šatínský, Dalibor (referee)
Author: Bc. Lucie Tetrevová Title: The application of chromatographic methods for separtion metabolites of cholesterol Language: czech The proposed diploma thesis deals with the development and validation of analytical method for free and esterified cholesterol separation from blood serum and adrenal tissue samples. This method is neccessary due to the different metabolism of free and esterified cholesterol in the body and it enables the research of these particular pathways using stable isotope tracers. The solid phase extraction method was chosen, optimised and validated with following results: precision values 2.14% for serum cholesterol ester, 6.98% for tissue cholesterol esters, 2.91% for serum free cholesterol, 7.48% for tissue free cholesterol, limit of detection 1 mol/l and linear range 1-25 mmol/l were found. The stability of the derivative was tested for 30 days at temperatures 4 C and -25 C. The derivative was found to be stable for the whole time period in both temperatures. Laboratory temperature was not tested due to a high volatility of the solvent used. The developed method was used in experimental studies evaluating the influence of dietary cholesterol and septic shock to cholesterol synthesis rate.
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Možnosti stanovení těkavých látek při výrobě piva
Rozsypalová, Markéta
The thesis discusses the possibilities of determination of chlorophenols in beer. A review of literature describes the complex properties of chlorinated phenols. Summarizes their physical, chemical and toxicological properties and characterizes their possible inputs into the environment. In the experimental part is the main theme of the determination of chlorophenols in samples of small breweries and beers called as home brew. These are various types of beer, in which the amount of chlorophenols determined using solid phase extraction (SPE) using gas chromatography with flame ionisation detector (FID). Another part of the experiment is aimed at demonstrating the formation of chlorophenols in beer by simulating conditions in the production of beer in the pilot plant. Chlorphenols amount of boiled beer is again determined by gas chromatography. Samples of beers are compared with each other in the content of chlorophenols. It is evaluated sectional representation monitored chlorophenols and their quantity in the analyzed samples of beer.
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The contents of selected persistent organic pollutants in soils at selected locations within Hradec Králové city
Bartošová, Barbora ; Száková, Jiřina (advisor) ; Radim, Radim (referee)
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants entering the environment from both natural and anthropogenic sources. The main anthropogenic sources are processes for the production of electricity and thermal energy, waste incineration, road traffic and motor vehicles, aircraft, boats, steam and diesel-electric locomotives and some industrial technologies. Besides these main sources there are still a number of others, less important, e.g. working with hot tar, asphalt pavement wear and tire during vehicle operation, coking plants, plants for processing of oil and some metals plants. An indispensable source in the cities as domestic solid fuel burning furnaces.
Although Hradec Králové do not belong to the cities with extreme levels of contaminants in the air, according to the Czech Hydrometeorological Institute, the levels of benzo(a)pyrene in the case of annual average concentrations exceeding the maximum allowable concentrations occurred here in 2013. While air quality is monitored regularly, the PAHs contents in soils and vegetation are not so. In our simple single experiment, we decided to assess the level of contamination of soil PAHs in the land of Hradec Kralove. It was collected 86 soil samples, in which were determined contents of 16 PAHs. Further, mushroom fruit samples that were found in the individual sampling points were analyzed.
The results showed large variability of PAHs content in the city, but did not indicate significant levels of contamination. The sum of all the determined PAU does not exceed the permissible value of the contents of these elements in soils. However, in several cases exceeding the maximum allowable content in soils with different PAHs, especially fluoranthene, anthracene and chrysene. Linear correlation analysis showed that the levels of all monitored PAHs are close correlations, which indicates that the entire spectre of PAHs in this case always originates from the same source. The PAH contents in mushrooms were in most cases below the detection limit of the determination, only rarely the detectable contents of individual PAHs were found (Cylocybe gambosa, Agaricus arvensis). We can speculate about the limited ability of fungi to accumulate PAHs or their ability to degrade these substances.
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Problems of Drug Residues Determination in Waste Waters
Lisá, Hana ; Márová, Ivana (referee) ; Hajšlová, Jana (referee) ; Kráčmar, Stanislav (referee) ; Vávrová, Milada (advisor)
The PhD thesis deals with determination of sulfonamides and tetracyclines in waste waters. Sulphonamides and tetracyclines are widely used in human and veterinary medicine. They are biologically active compounds and block biological processes in waste water treatment plants. These substances can influence microorganisms in the environment. Sulphonamides and tetracyclines induce resistance in low concentrations. These substances penetrate into the environment from industrial production, use in households, hospitals, veterinary clinics and fish farms. Tetracyclines and sulphonamides were determined in waste water from waste water treatment plants in Brno. Optimalization of extraction of these compounds was performed by solid phase extraction. Sulphonamides and tetracyclines were determined by liquid chromatography coupled with diode array detector or coupled with mass spectrometry. Optimalized method was used for determination tetracyclines and sulphonamides in waste water from waste water treatment plants in Brno in years 2007 – 2010.
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Residual analysis of botanical pesticides using separation methods
Medová, Tereza ; Nový, Pavel (advisor) ; Táborský, Jan (referee)
Botanical pesticides that include essential oils are considered to be one of the possible alternatives to synthetic chemical pesticides in plant protection, therefore it is important to develop our knowledge about these natural substances. This Master thesis comprises the residual analysis of selected compounds contained in Cymbopogon citratus Stapf, Pelargonium graveolens L'Hér and Thymus vulgaris L. essential oils after their application on plant material. The chemical composition of each essential oil was initially analyzed by gas chromatography - mass spectrometry (GC - MS). This analysis helped to select three representative compounds for each essential oil and the residues were determined for each of them. Citral, Geranyl-acetate and Limonene were selected for C. citratus, b-Citronellol, Geraniol and Linalool for P. graveolens and Thymol, p-Cymene and g-Terpinene for T. vulgaris. Two new methods to determine botanical pesticide residues were developed. The new methods should provide faster and easier analysis. One of them uses Solid Phase Microextraction method (SPME). The other one, that was applied in the residual analysis, uses hexane for extraction. The results show that essential oils' persistence is very low on the surface of plant material, which would imply they are environmentally friendly and safe for human health assuming correct manipulation with these substances.
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