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Acetic acid as a side product of metabolism of PHA producing bacteria
Ryšavá, Lenka ; Obručová,, Hana (referee) ; Obruča, Stanislav (advisor)
The Bachelor´s thesis deals with experimental study production of acetic acid as a side product of metabolism of PHA producing bacteria. Experimental part of work deals with the impact of used substrate on the biomass growth, PHA production and the formation of organic acids in strains that can potentially find their application in industrial production of PHA – Cupriavidus necator H16, Burkholderia sacchari, Burkholderia cepacia. The highest yields of organic acid have been reached on Burkholderia sacchari when cultivated on mineral medium with glucose. The last part of the thesis is focused on determining the viability of micro organisms using flow cytometry where bacteria Cupriavidus necator H16, Burkholderia sacchari, Burkholderia cepacia were exposed to selected stress factors.
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Application of polymer membranes in microextraction techniques for analysis of biological samples
Ryšavá, Lenka ; Diviš, Pavel (referee) ; Dvořák, Miloš (advisor)
The thesis presents experimental study on application of various polypropylene membranes (with different thicknesses, porosity and pore size) for direct coupling of membrane microextractions with capillary electrophoresis. No comprehensive study, which describes effect of these membrane parameters on extraction recoveries, was published in the past. Previous scientific works prefer application of 100 µm polypropylene membranes for their easy handling and satisfactory extraction efficiencies. Experimental part includes examination of selected polypropylene membranes as supported liquid membranes in-line coupled to capillary electrophoresis for analysis of basic drugs from complex samples. Membranes with three thicknesses (25, 100 and 170 µm) were tested. The highest extraction recoveries were achieved for the 25µm thick polypropylene membrane. Various pH conditions of donor and acceptor operational solutions were examined for extractions from real complex matrices (urine, plasma). The optimal extraction conditions were 10 mM NaOH as donor phase and 10 mM HCl as acceptor phase. 25µm membranes offer higher extraction recoveries, reduced consumption of organic solvents for membrane impregnation, similar mechanical stability and similar clean-up ability compared to thicker polypropylene membranes.
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Application of capillary electrophoresis for dried blood spots analysis
Ryšavá, Lenka ; Tůma,, Petr (referee) ; Petr,, Jan (referee) ; Kubáň, Pavel (advisor)
Blood samples are normally analyzed in the form of plasma or serum. As an alternative for blood sampling and storage, dry blood spots (DBS) have been increasingly used in recent years. In the actual dissertation thesis, DBS are processed and analyzed by capillary electrophoresis (CE), which has been very rarely used for DBS analysis in the past. However, CE offers several features, given by the recent development of this technique, not available for standard analytical methods. Direct coupling of microextraction techniques to CE and simultaneous determination of analytes present in complex matrices leads to minimization or elimination of deficiencies in DBS analysis, for example the effect of sample matrix, hematocrit effect, and inhomogeneous distribution of analytes in DBS. In the simplest possible arrangement, sample treatment, injection, and analysis of DBS samples are achieved exclusively by a single commercial CE instrument. Application of newly developed fully soluble materials for DBS sampling offers better extraction efficiency and more accurate quantitation. The developed concepts include new methods for efficient treatment of DBS samples and their direct analysis without the need for operator intervention and they provide sufficient selectivity and sensitivity for the determination of important analytes not only in DBSs but also in other complex samples.
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Rapid fabrication of microextraction devices for direct determination of basic drugs in dried blood spots
Ryšavá, Lenka ; Přikryl, Jan ; Dvořák, Miloš ; Malá, Zdeňka ; Kubáň, Pavel
A 3D-printed cutter was designed for rapid fabrication of supported liquid membrane (SLM) extraction devices compatible with injection system of commercial capillary electrophoresis (CE) instrument. Low cost and single use microextraction devices were fabricated from polypropylene micropipette tips with high precision (dimension variances ≤ 3% RSD). Practical applicability of the microextraction devices was examined by in-line coupling of SLM extractions to CE for direct analysis of four model basic drugs in dried blood spot extracts. Excellent repeatability of the hyphenated technique was achieved (RSD ≤ 7.6%), transfer of analytes across the SLM ranged from 23 to 73% and limits of detection were as low as 0.07 μg/mL.
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Application of polymer membranes in microextraction techniques for analysis of biological samples
Ryšavá, Lenka ; Diviš, Pavel (referee) ; Dvořák, Miloš (advisor)
The thesis presents experimental study on application of various polypropylene membranes (with different thicknesses, porosity and pore size) for direct coupling of membrane microextractions with capillary electrophoresis. No comprehensive study, which describes effect of these membrane parameters on extraction recoveries, was published in the past. Previous scientific works prefer application of 100 µm polypropylene membranes for their easy handling and satisfactory extraction efficiencies. Experimental part includes examination of selected polypropylene membranes as supported liquid membranes in-line coupled to capillary electrophoresis for analysis of basic drugs from complex samples. Membranes with three thicknesses (25, 100 and 170 µm) were tested. The highest extraction recoveries were achieved for the 25µm thick polypropylene membrane. Various pH conditions of donor and acceptor operational solutions were examined for extractions from real complex matrices (urine, plasma). The optimal extraction conditions were 10 mM NaOH as donor phase and 10 mM HCl as acceptor phase. 25µm membranes offer higher extraction recoveries, reduced consumption of organic solvents for membrane impregnation, similar mechanical stability and similar clean-up ability compared to thicker polypropylene membranes.
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Strategický plán mikroregionu
Ryšavá, Lenka
The diploma thesis aimed at "Strategic plan of the microregion Moravskokrumlovsko" focuses on forming a strategic development plan for the microregion Moravskokrumlovsko. The thesis is divided into several parts. The first part, a literature search, includes a determination of basic terms and a method for forming the strategic plan. The second part of the thesis, the work itself, contains the socioeconomic analysis of the microregion Moravskokrumlovsko which is evaluated through the SWOT analysis. The thesis also includes an interrogatory survey. The strategic plan and the key areas that should be developed in the microregion in the future are proposed. Finally, there are evaluated total results of the analysis and possible recommendations are presented.
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Acetic acid as a side product of metabolism of PHA producing bacteria
Ryšavá, Lenka ; Obručová,, Hana (referee) ; Obruča, Stanislav (advisor)
The Bachelor´s thesis deals with experimental study production of acetic acid as a side product of metabolism of PHA producing bacteria. Experimental part of work deals with the impact of used substrate on the biomass growth, PHA production and the formation of organic acids in strains that can potentially find their application in industrial production of PHA – Cupriavidus necator H16, Burkholderia sacchari, Burkholderia cepacia. The highest yields of organic acid have been reached on Burkholderia sacchari when cultivated on mineral medium with glucose. The last part of the thesis is focused on determining the viability of micro organisms using flow cytometry where bacteria Cupriavidus necator H16, Burkholderia sacchari, Burkholderia cepacia were exposed to selected stress factors.
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