Constantly increasing consumption of drugs leads to the increase of the concentration of residues in various environmental compartments. For this reason the drug residues are nowadays classified as significant environmental contaminants. The drug residues enter the soil along with the application of sewage sludge either as a fertilizer or during fertilizing by animal excrements excreted after drug administration, eventually by direct contamination from treated grazing animals. The presence of these substances in soil has got negative impact on the sensitive soil ecosystem. The diploma thesis aims to evaluation of possible soil contamination with selected drugs through ecotoxicity tests. For this purpose sulfonamide chemotherapeutic agents, which are often used in veterinary medicine, were chosen, specifically sulfamethoxazole and sulfamethazin. Eartworms (Lumbricus) were selected as suitable test representative soil organism. In the experimantal part of this work the avoidance test according to ISO 17512-1 and modified bioaccumulation test according to OECD 317 methodology were performed. Bioaccumulation substances in oligochaetes, for which the artificial soil was prepared according to OECD 207 methodology, which was subsequently contaminated with above specified drugs. Although according to the results of determination of these substances there is no significant bioaccumulation in eartworms‘ tissue, these organisms are very responsive to the presence of contaminants in the soil environment. This is evident from the results of avoidance tests.

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.

This diploma thesis deals with the determination and ecotoxicological assessment of artificial sweeteners. The diploma was focused on monitoring the most commonly used sweeteners, which includes acesulfam-K, cyclamate, saccharin and sucralose. These sweeteners were analyzed by liquid chromatography with mass spectrometry, using solid phase extraction for their concentration. The samples for their determination were taken from wastewater treatment plant in Modrice. Artificial sweeteners are synthetically produced substances, which is the main reason for their occurrence in waste water. Some of them are not efficiently removed from the wastewater treatment plant and penetrate into the environment. Analysis of the samples was found, that sucralose penetrates into the environment. The ecotoxicological effect of the sweeteners was evaluated via aquatic organisms, namely on the freshwater crustaceans Thamnocephalus platyurus and Daphnia magna, on the water plant Lemna minor, and the battery of tests was supplemented by a representative of the destructors Vibrio fischeri. For all sweeteners, LC20, EC20 and IC20 values were determined. For some sweeteners, LC50, EC50 and IC50 values were also determined. During testing, saccharin represented the highest toxicity.

One theory concerning the invasiveness of exotic plants suggests that they exude allelopathic compounds that are novel in areas being invaded. Here, I tested for allelopathic effects of root exudates of the invasive plant Heracleum mantegazzianum (giant hogweed) in a series of experiments both in vitro and in the garden, and compared them with effects of its native congener H. sphondylium (common hogweed) and two less related native species. Moreover, I addressed intraspecific variability of allelopathy by comparing effects of 41 different maternal lines of H. mantegazzianum sampled from several areas and populations within the Czech Republic. Not only the effects, but also the identity of allelopathic compounds was investigated. In order to test also for the interaction of allelochemicals with soil microorganisms, I analysed the composition of soil microbial communities expressed as phospholipid and neutral lipid fatty acids (PLFA and NLFA, respectively) together with nutrient conditions and light availability at invaded sites in the Czech Republic. The results show that H. mantegazzianum is able to exert phytotoxic effects in vitro as well as in the garden experiment; however, its effects did not differ from the inhibition caused by the native species tested. Variation partitioning among areas,...

Normotensive hydrocephalus (NPH) is a neurodegenerative disease that occurs mainly in patients of high age. The disorder causes accumulation of cerebrospinal fluid (CSF), which leads to enlargement of ventricles and pressure exerted on cerebral structures. Clinical manifestations (gait disorders, development of dementia, incontinence) can be easily confused with symptoms of other neurodegenerative diseases; unlike other such disorders, however, NPH can be treated by surgery, if diagnosed in time. Patients are indicated for the procedure by a lumbar drainage test. There are currently no reliable laboratory biomarkers known that could be the basis of NPH diagnostics. In the past, steroids proved to be linked to neuronal activity in neurodegenerative diseases with the help of specific diagnostic markers. An instrumental method has been developed for the purposes of this thesis and it was used to gauge the level of certain steroids in CSF in a sample group of NPH patients and a control sample group of healthy individuals. A significant difference has been found in levels of aldosterone and cortisone. Aldosterone was higher in NPH sufferers, while cortisone levels were higher in the control group. It is crucial to differentiate patients with NPH from patients with similar clinical manifestations during...

The aim of this work was development of high-performance liquid chromatography method with DAD detection for determination of nicotine in tobacco. Standard operating procedure used by World Health Organization was chosen as comparison of the developed method. Optimized high performance chromatography method is suitable for determining nicotine in tobacco. Limit of detection for this method was 0,0003 mg/ml and limit of quantification was 0,0010 mg/ml. Optimization of preparation of samples was significant part of this thesis. Sample preparation procedure was made substantially easier in comparison to other commonly used methods. Nicotine content was determined from real tobacco leaves samples, cigarette tobacco filler, nicotine cartridge for electronic cigarettes and pipe tobacco. Satisfactory relative standard deviation was achieved for all types of samples. Next part of this thesis focused on study of determining polyphenols using high-performance liquid chromatography with diode array detector. Chosen analytes were chlorogenic acid, caffeic acid, rutine, scopoletine and quercitrine. Among the five tested analytes, the highest sensitivity was achieved for chlorogenic acid and caffeic acid. All of the analytes achieved low limits of detection and quantification. Key words Liquid chromatography,...

A simple method has been developed to determine amino acid loading in solid phase peptide synthesis. The method is applicable for the most common type of synthesis, which use FMOC as protective group and piperidine as a deprotecting agent. Both products of deprotection reaction are separated by HPLC and determined using an UV detector; an internal standard is added. The method gives true values that have been verified by an independent method. The RSD is 1.52%. The method is more accurate than the published methods and allows the determination from the waste of synthesis. The employing of the internal standard eliminated the necessity of dilution accuracy or known volumes.

This diploma thesis deals with the determination of glucosinolates in plant material by capillary electrophoresis and high performance liquid chromatography. The chemical structure, biosynthesis, degradation, and also biological effects of glucosinolates are described. One part of this work also deals with the methods, which glucosinolates in plant materials were determined by. The experimental part describes the separation of intact glucosinolates by capillary electrophoresis and high performance liquid chromatography. Two plant materials were available for the determination of glucosinolates, namely lyophilized rapeseed leaves and broccoli juice. Micellar electrokinetic chromatography using a cationic surfactant was used to determine intact glucosinolates by capillary electrophoresis. After finding the optimal conditions for the separation of intact glucosinolates, it was found that it is impossible to determine these substances in plant samples. The reason was interference from the matrix, which interfered with this determination. While using high performance liquid chromatography under optimal conditions, some of the intact glucosinolates were identified in a rapeseed plant sample. Furthermore, the calibration dependencies of individual glucosinolates were obtained and the recovery and...

The thesis on the topic Soil pollution by phthalic acid esters in Czech Republic solves the problem of soil pollution by dibutyl phthalate (DBP) and di (2-ethylhexyl) phthalate (DEHP). This thesis joined the annual monitoring conducted by Central Control and Testing Institute of Agriculture. The monitoring of contaminants was carried in twelve regions of the Czech Republic. The samples do represent different types of culture farmland. They were formed by arable land, permanent grassland and hop-garden. Before the analysis, the samples were extracted with acetone-hexane mixture with 1:1 ratio, and ultrasound. Samples were analyzed by liquid chromatography with UV detection. The measured concentrations for the year 2016 ranged in DBP from 0.07 to 0.21 mg.kg-1 dry weight, at DEHP from 0.19 to 1.08 mg.kg-1 dry weight, esters together from 0.13 to 0.65 mg.kg-1.

Submitted master thesis summarizes current knowledge about biologically active sub-stances contained in soybeans (Glycine max L. Merrill). Mainly deals with problematics of isoflavones, mechanism of origin and effects on human healht. Then the theoretical part summarizes mainly modern instrumental technics using for determination of isofla-vones. The experimantal part is concentrated on identification of soy isoflavones gathered from eight varieties (Brunensis, Bohemians, Naya, Moravians, Silesia, Toutatis, Korus, Kofu) of soybeans cultivated in Czech Republic. For detection of free, conjugated and total isoflavones was used high performance liquid chromatography with mass spectro-meter (LC-MS/MS)