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Post-column derivatization in HPLC for analysis of amino acids
Vaňkátová, Petra ; Coufal, Pavel (advisor) ; Sobotníková, Jana (referee)
Proteinogenic amino acids are the basic structural building units of proteins. Their analysis is important in many fields, especially in medicine. This thesis deals with one of the methods of derivatization of amino acids in order to increase their absorption in the UV - the post-column derivatization using copper (II) oxide. It is quick, robust and easy-to-use method. Following the nowadays trend in the analysis of free amino acids, this thesis is focused on the HILIC mode separation environment.
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HPLC determination of gallic acid as a possible product of enzymatic reaction of shikimic acid, NADP+ and SDH.
Smolejová, Jana ; Červený, Václav (advisor) ; Nesměrák, Karel (referee)
This diploma thesis deals with the development of an HPLC method for the determination of selected compounds participating in enzymatic reaction leading to the formation of gallic acid. The analysed reaction mixture contains the following reagents: shikimic acid, NADP+ and shikimatedehydrogenase (SDH) extracted from parsley; the presumed product of the reaction is gallic acid. Two chromatographic methods for the determination of the above mentioned compounds were developed using C18 HPLC column and porous graphitic carbon Hypercarb column. Molecular absorption spectrometric detection in the UV range was used in all measurements. Separation on the C18 column was found particularly suitable for analysing the composition of the end products of the reaction. Because of the NADP+ and shikimic acid peak overlap it is necessary to observe absorbance at 212 and 260 nm. Shikimic acid and NADP+ can be quantified due to the fact that shikimic acid does not absorb at 260 nm while NADP+ absorb radiation at both wavelengths. Separation via Hypercarb column was found particularly suitable for analysing the process of the reaction; additional products or intermediates can be seen in chromatograms compared to the C18 method. Determination with Hypercarb column is characterized by higher sensitivity and lower limit...
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Post-column derivatization in HPLC for analysis of amino acids
Vaňkátová, Petra ; Coufal, Pavel (advisor) ; Sobotníková, Jana (referee)
Proteinogenic amino acids are the basic structural building units of proteins. Their analysis is important in many fields, especially in medicine. This thesis deals with one of the methods of derivatization of amino acids in order to increase their absorption in the UV - the post-column derivatization using copper (II) oxide. It is quick, robust and easy-to-use method. Following the nowadays trend in the analysis of free amino acids, this thesis is focused on the HILIC mode separation environment.
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Electrophoresis in short capillary with dual detection.
Kaliba, David ; Opekar, František (advisor) ; Jelínek, Ivan (referee)
Capillary zone electrophoresis is an analytical method frequently used in many laboratories for solving various analytical problems. This diploma thesis describes one of many applications of capillary zone electrophoresis using a unique laboratory apparatus composed of a short capillary and dual conductivity/UV detector placed in one detection point of the separation capillary. In the first part of this thesis, the laboratory apparatus was tested by the separation of small inorganic and organic ions. Sodium, potassium, tyramine and histidine ions were used to test the two parts of the dual detector. Experimentally obtained mobilities of these ions were compared with those calculated from the tabulated values. In the second part, the apparatus was used for determination of analytes in samples with more complex matrixces, pharmaceuticals Acylcoffin and B-komplex produced by Zentiva, a.s. One analyte was chosen from each pharmaceutical preparation for determination of its concentration in the preparation, caffeine from Acylcoffine and thiamine from B-komplex. The concentrations were calulated using three different calibration methods and the experimentally obtained values were compared with those specified by the pharmaceuticals producer. Key words: capillary electrophoresis; short capillary;...
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Labio, a.s. – research study of gas chromatograph enhancement
Divoký, Martin
This project aimed at testing and development of a UV gas chromatograph Labio G11. A spectrometer from the chromatograph was modified in order to increase nitrogen gas flow in the device and speed at which the inner of the spectrometer is filled with nitrogen (that doesn’t absorb in UV). This allowed decreasing startup time of the spectrometer. Additionally, sensitivity of the spectrometer in region below 150 nm was tested and ways to increase this sensitivity were proposed.
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