The Diploma Thesis is focused on the effect of the preparation Terra-Sorb which contains free amino acids, on the yield and the quality of choosen field crops. The main attention is pursued especially on the use of nitrogen and aminoacids in the plant nutrition. It's analyzed mainly in the literature review which is also focused on the intake of the nutrient and foliar nutriet of the field crops. The practical part is focused mainly on the verification of the effect of the stimulator Terra-Sorb. It's effect was verified during annual and perennial small plot trials on the location in Velká Bystřice. There were verified the different applications of the stimulator Terra-Sorb in the winter wheat, sugar beet and spring barley. There were evaluated yield and quality of each crops.

This thesis is dealing with effect of supplement limiting aminoacid histidin, that was applied as duodenal infusion, on yield and composition of high production dairy cows milk. The theoretical part describes overview about milk composition and metabolism of dairy cows. There are described nutrition requirements of dairy cows for esential aminoacids. In this part are also presented principles of analytical methods used for analysis of milk. For determination of basic components of milk were used infrared spectroscopy and enzymatic-conductometry. Aminoacid profile of casein was determined by ion exchange liquid chromatography. In the experimental part is described method of experiment. There is described technique of acid and oxidative hydrolysis of lyofilized casein and technique of determination aminoacids in this hydrolyzated casein. Results of analysis were interpreted and discussed with literature. This diploma thesis was made as a cooperation between the Institute of Food Science and Biotechnology, the Faculty of Chemistry, University of Technology, Brno and the Research Institute for Cattle Breeding, Department of Animal Physiology and Nutrition, Pohorelice.

This work is focused on interaction between hyaluronan (Hya) and amphiphilic molecules. Using fluorescent probes method were carried out screening of the interaction of various surfactants with Hya. For further examination was chosen cetyltrimethylammonium bromide (CTAB). Interactions in this system have also been studied using fluorescence correlation spectroscopy (FCS), and additionally were performed initial tests of the cytotoxicity of the system. As an alternative CTAB were also used hydrophobically modified amino acids. Results showed their self-aggregation, but their interactions with hylauronanem was not as strong as with CTAB. The interaction of gel systems Hya-CTAB have also been studied, and was found to contain a hydrophobic domain capable solubilize hydrophobic substance. Thermogravimetric have been established that are able to contain up to 98% wt. water and rheologically were characterized their mechanical properties. These are tunable due to molecular weight (MW) of the hyaluronate. With increasing value of MW increases mechanical strength of the gel and decreases its fluidity.

This dissertation thesis composes of two parts; the first part focus on the characterization of trypsin, enzyme frequently used in proteomic research for the investigation and identification of protein sequences, and its peptide digestion kinetics. The second part is aimed to the enantioseparations of biologically active compounds. First part of this project focus on tryptic digestion of synthetic peptides and the development of HPLC method for the identification of synthetic peptides and their fragments. Using the in-solution digestion and HPLC method, relative kinetic constants were determined for problematic sequences. Amino acids responsible for the decrease in trypsin catalytic activity and their location towards the cleavage site were studied. Certain slight exopeptidase activity of trypsin was noted, especially at the end of peptide chain. Furthermore, three columns with immobilized trypsin used in HPLC were compared concerning their catalytic activity. The immobilization of enzymes on solid support is used to elevate the amount of enzyme present during digestion and to assure better repeatability and reproducibility of obtained results. Activity of a new trypsin column synthesized at the University of North Carolina at Chapel Hill was compared to two commercially available trypsin columns....

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.

In this work, an enantioselective HPLC method with UV and fluorimetric detection was developed and subsequently optimized for chiral separation of four aminoacids (D/L-alanine, D/L-valine, D/L-leucine, D/L-isoleucine) in native and particularly in derivatized form with an emphasis on enantioseparation of D-analogues. Retention and enantioseparation behavior of studied analytes was investigated on three chiral stationary phases based on teicoplanin (Chirobiotic T, Chirobiotic T2) and teicoplanin aglycone (Chirobiotic TAG). At the Chirobiotic T column, enantioseparations of underivatized aminoacids were performed with UV detection at 205 nm in the mobile phases methanol/water with different volume ratios. Baseline separation of L- and D-forms was achieved, however, the sensitivity of detection was very low. In order to increase detection sensitivity, derivatization of aminoacids was performed using 9-fluorenylmethyl chloroformate (FMOC-Cl) and the derivatization procedure was monitored on Chirobiotic T column with fluorimetric detection (λEx = 254 nm, λEm = 314 nm) in a buffered mobile phase methanol/0.5% TEAA buffer, pH 6.0 40/60 (v/v). In terms of derivatization, volume ratio D/L-aminoacid/derivatization agent 1/1 with ten times higher concentration of derivatization agent was found to be the most suitable....

Anorexia nervosa (MA) is serious psychiatric disease associated with significant mortality. One of the main causes of mortality in these patients is cardiac complications. In 2003 - 2010 we examined 30 patients with established diagnosis of MA. Routinely used nutritional parameters are normal even in severe malnutrition in MA patients and we can ́t use them to diagnose or monitor nutritional status. Therefore we evaluated plasma aminoacids levels to detect minor changes of protein metabolism. According to our results we found mild hyperaminoacidemia and significant elevation of Gly/Val and Phe/Tyr ratio. In the second part of our research we performed non invasive examination s in risky patients with AN to consider possible cardiac complications. Results of ECG and echocardiography confirm results of the previous published studies. Original finding is significant decrease in flow mediated dilatation of brachial artery and its normalization after realimentation. We found also in ambulatory blood pressure monitoring significant decrease in blood pressure during the active period which was the consequence of changes in the heart rate variability. Decrease of the blood pressure in patients with AN in standing position was associated with increase of HF power in supine position and particularly with the lack of...

The first aim of this work was to develop copper modified carbon felt electrode for detection of amino acids, which are not electrochemically active on ordinary carbon electrodes. Phenylalanine solution at a concentration 1.0 mmol dm-3 was chosen as the testing solution. Electrode modified with electrochemical deposition of copper from mixture of copper(II) sulphate and sodium sulphate provided very low responses which were decreasing during first measurements, apparently because of insufficient amount of copper. Therefore, further experiments were performed using copper microparticles as a modifier. Copper microparticles activated in nitric acid at a concentration 80 mmol dm-3 were applied at carbon felt by several techniques and in different parts of the felt, i.e. by stirring the felt in microparticles suspension, by dripping of the suspension to the part of the felt in contact with capillary (proximal), between two parts of the carbon felt (sandwich) and at a part of the carbon felt in contact with electrolyte in a measuring cell (distal). Electrodes modified in the distal and in the sandwich arrangement were chosen as the best ones. In the last part, calibration dependences for phenylalanine in concentration range from 0.025 mmol dm-3 to 1.0 mmol dm-3 were measured on the last two electrodes....

This diploma thesis deals with the study of adsorption of low-molecular components of organic substances produced by phytoplankton - AOM (Algal Organic Matter) on AC (Activated Carbon) during drinking water treatment. For the experimental purpose in this thesis, there were used selected amino acids as low molecular substances of AOM that are difficult to remove by conventional water treatment process by coagulation. As adsorbent, there was used a detailed characterized of granulated activated carbon (GAC) - Filtrasorb TL 830 (FTL830) which is intended directly for the purpose of water treatment. There were realizing the equilibrium batch adsorption experiments with three different model amino acids - arginine (Arg), phenylalanine (Phe) and aspartic acid (Asp). There was investigated the efficiency of removing amino acid depending on the solution temperature and pH. Results of the adsorption experiments have shown that the temperature affects the adsorption efficiency. Adsorption is essentially described as an exothermic process but the adsorption of Arg and Phe from an aqueous solution to GAC occurs more efficiently at higher temperatures. It means that the adsorption is the endothermic process. In the case of Arg adsorption, the temperature was found to influence adsorption efficiency less than...

(EN) Proteinogenic amino acids are key components of living organisms. Thus, the latest metabolomics research has focused on developing fast and sensitive methods for the determination of amino acids. In this context, this thesis contains two studies describing development of high-performance separation techniques for the quantification of amino acids. In the first study, a capillary electrophoresis method was developed for the determination of free amino acids in tobacco plants, particularly focusing on optimizing the extraction of amino acids from solid plant materials. The extraction procedure was optimized using design of experiments (DoE) to obtain the highest possible extraction yield of amino acids. Factors such as volume and concentration of the extraction solvent (hydrochloric acid) were assessed as the most significant. Subsequently, the optimal values of these factors were determined using response surface methodology (RSM). Lastly, proteinogenic amino acids were quantified using capillary electrophoresis with contactless conductivity detection and calibration with internal standard, which improved the precision of the method. The second study aimed at developing a supercritical fluid chromatography method for the determination of free proteinogenic amino acids in human plasma. The most...