This thesis presents a thermal gradient gas chromatography (TGGC) system that is implemented on a micro-scale. The GC column is approximately 20 cm long and is fabricated out of silicon with 21 nickel thin-film heaters evenly placed along the length of the column. Computational heat transfer models using ANSYS Mechanical APDL predict heating and cooling rates up to 32,000 deg C/min and 3,600 deg C/min, respectively. These results are verified through testing an experimental silicon channel. A PI controller which uses resistance measurements to calculate thin-film temperature is used for obtaining dynamic thermal gradient control. This controller is shown to possess a characteristic rise time of approximately 0.3 seconds with less than 4% overshoot and precision to within less than a degree. These characteristics present this system as a highly favorable candidate for a micro-GC column with resolution similar to that of conventional GC.

<p>The aim of this study is to identify metabolic transformations in breast cancer through urinary volatile organic compounds in mammary pad or bone tumor mice models. Subsequently, it focuses on investigating the efficacy of therapeutic intervention through identified potential biomarkers. Methods for monitoring tumor development and treatment responses have technologically advanced over the years leading to significant increase in percent survival rates. Although these modalities are reliable, it would be beneficial to observe disease progression from a new perspective to gain greater understanding of cancer pathogenesis. Analysis of cellular energetics affected by cancer using bio-fluids can non-invasively help in prognosis and selection of treatment regimens. The hypothesis is altered profiles of urinary volatile metabolites is directly related to disrupted metabolic pathways. Additionally, effectiveness of treatments can be indicated through changes in concentration of metabolites. In this ancillary experiment, mouse urine specimens were analyzed using gas chromatography-mass spectrometry, an analytical chemistry tool in identifying volatile organic compounds. Female BALB/c mice were injected with 4T1.2 murine breast tumor cells in the mammary fat pad. Consecutively, 4T1.2 cells were injected in the right iliac artery of BALB/c mice and E0771 tumor cells injected in the tibia of C57BL/6 mice to model bone tumor. The effect of two different modes of treatment: chemical drug and mechanical stimulation was investigated through changes in compound profiles. Chemical drug therapy was conducted with dopamine agents, Triuoperazine, Fluphenazine and a statin, Pitavastatin. Mechanical stimulation included tibia and knee loading at the site of tumor cell injection were given to mice. A biological treatment mode included administration of A5 osteocyte cell line. A set of potential volatile organic compounds biomarkers differentiating mammary pad or bone confined tumors from healthy controls was identified using forward feature selection. Effect of treatments was demonstrated through hierarchical heat maps and multivariate data analysis. Compounds identified in series of experiments belonged to the class of terpenoids, precursors of cholesterol molecules. Terpene synthesis is a descending step of mevalonate pathway suggesting its potential role in cancer pathogenesis. This thesis demonstrates the ability of urine volatilomics to indicate signaling pathways inflicted in tumors. It proposes a concept of using urine to detect tumor developments at two distinct locations as well as to monitor treatment efficacy.</p><br>

<p>Solid Phase Microextraction (SPME) is a popular sampling technique that can be paired with Gas Chromatography/Mass Spectrometry (GC-MS). SPME-GC-MS is used in forensic chemistry due to its simplification of the sample preparation process. Headspace-Solid Phase Microextraction (HS-SPME) is a technique where the sample is heated to generate volatiles in the headspace of the vial. A SPME fiber is then inserted into the vial and the compounds in the headspace will bind to the fiber. Total Vaporization- Solid Phase Microextraction (TV-SPME) is a technique that is derived from the HS-SPME technique. </p><p>In Chapter 1, the critical comparison of HS-SPME and TV-SPME is discussed. Samples including marijuana, essential oils, and CBD oil were utilized to compare the two techniques. The compounds of interest in marijuana are the three main cannabinoids: cannabinol (CBN), cannabidiol (CBD), and tetrahydrocannabinol (THC). The sample preparation and GC-MS parameters were kept the same for all samples to determine which SPME technique works best for these sample types and yielded the greatest sensitivity. It was found that HS-SPME shows greater sensitivity with CBN and equivalent sensitivity with essential oils, THC and CBD. </p><p>In Chapter 2, the detection of synthetic cannabinoids utilizing liquid-liquid injection as well as HS-SPME and TV-SPME is discussed. The detection of these compounds is important because this type of drug has become more prevalent in the United States because they can be chemically altered slightly so they still have the effects of a drug but can evade drug legislation. The detection of synthetic cannabinoids using liquid injection was found to be successful but detection using HS-SPME and TV-SPME was found to be unsuccessful. </p>In Chapter 3, the analyses of real and artificial saliva utilizing HS-SPME and TV-SPME is discussed. Determining the compounds present in real saliva and artificial saliva will be of importance for future research into determining if the presence of drugs in saliva can be analyzed with these techniques. The analyses of real and artificial saliva were found to be successful using HS-SPME, without derivatization, and TV-SPME, with and without derivatization. Many of the compounds present in the real saliva were detected and were confirmed to be compounds regularly found in saliva by other scientific literature.

This diploma thesis deals with the monitoring of chemical changes in ground coffee stored in various containers and then it deals with the determination of water, moisture and aromatic profile of coffee in individual containers, namely a can, a paper bag, a bag with a polyethylene liner, a bag from low density polyethylene with aluminium liner (LDPE + Al) and a glass container. At the beginning of the experiment it was found that the water content in the coffee was 4.72 ± 0.10 % and the moisture content was 9.47 ± 0.01 %. After the original sample was measured, the coffee was poured into 5 packages. At the end of the experiment, a can was chosen as the most suitable package where the lowest water content was measured. The water content was determined to be 6.51 ± 0.11 % and the moisture content was 9.93 ± 0.01 %. However, the differences between the other packages, apart from the paper bag, were very small, and thus in general it can be said that the can, the LDPE + Al package, the bag with the PE liner and the glass container are suitable for storing ground coffee. A total of 44 aromatics were identified when determining the aromatic profile in coffee by HS-SPME-GC-MS. The most represented groups of substances were furans and pyrazines. During storage, there was some loss of substances due to 2 mechanisms. The first of them is evaporation and the second one is the oxidation reactions themselves. Most aromatic substances have evaporated / degraded in paper packaging, where this large decrease is related to its barrier properties. A glass container was chosen as the most suitable packaging with the least loss of aromatic substances.

The diploma thesis is focused on the issue of the presence of POPs in the surface water and sediments. Monitored compounds were chosen from the group of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDE). Samples were collected in selected localities in the Slovak and the Czech Republic. Selected analytes were extracted with hexane from the aqueous matrix. Analytes were isolated by the ultrasonic extraction using a solvent mixture of hexane-dichlormethane from the solid matrix and the resulting extract was cleaned up by the column chromatography filled with the florisil and the silica gel. Target compounds were analysed by GC/ECD finally. Obtained results show the level of contamination in selected localities in the Slovak and the Czech Republic.

This thesis is focused on determination of fatty acids in processed and/or processed cheese analogues using gas chromatography with FID detection. Characterization of fatty acids, various methods for lipid extraction and the possibilities of determination of fatty acids are described in the theoretical part. Simple and rapid method for the determination of fatty acids in natural and/or processed cheese was optimized and validated in the experimental part. The modified Folch method (by mixture of chloroform - methanol) was selected for lipid extraction from the samples. The standard method (ISO 5509), using a methanol solution of potassium hydroxide, was selected for lipid esterification. Type of the stationary phase and parameters of the column, flow of carrier gas, temperature of the injector, injection technique, temperature of the detector and temperature program were optimized. Calibration curves were constructed from five calibration points (in the concentration range of 0.01 - 60 g.ml-1), all correlation coefficients R2 were higher than 0,99. The limits of detection and quantification of individual fatty acids ranged from 0.002 - 6 g.ml-1. The repeatability of the method was good, all the relative standard deviations of the peaks were below 9 %. The optimized and validated method was applied to selected samples of processed cheese analogues. In total five types of analogues containing different kinds of fat (butter, apricot, linseed, black currant and grape seed oils) were analyzed. In samples 34 fatty acids were identified based on a comparison of retention times of standards.

- 4 - Abstrakt The aim of this diploma thesis was to determine the possibility of identifying the protein and lipid additives, such as blood, gelatine, curd, lard and eggs, in the model samples of lime based mortars using Raman spectroscopy and gas chromatography- mass spectrometry. It was proved that the method of Raman spectroscopy can determine the presence of organic additives in the order of one weight percent. The gas chromatography enables to detect lipid additives from the order of hundredths weight percent. Unambiguous resolution of protein additives can be done in the samples with the concentrations up to the order of one weight percent of the additives.

Chemical substances are transferred between bumblebees of the species Bombus terrestris during their mating behavior. These substances are present mainly on the cuticle. The aim of this thesis was to identify and quantitatify differences in the amount of the compounds among defined groups of bumblebees - mated, non-mated and attempting to mate. Cuticular extracts were obtained by washing of the whole bumblebees bodies in hexane. These extracts were analyzed by using a two-dimensional gas chromatography with the time of fly mass spectrometr (GCxGC-TOF/MS). Cuticular hydrocarbons were identified and semi-quantified. Double bounds locations were determined in heptacosene and nonacosene. Five compounds were differed in the examined groups - β-springene, α-springene, dihydrofarnesyl-oleate, palmityl-palmitate and palmityl- palmitoleate. By comparing of the amount of these compounds differences between defined groups of bumblebees were determined. Analysis of the cutilar extracts showed that mated and non-mated bumblebees can be distinguished. Furthermore, there were statistically significant differences in the amounts of identified compounds in all defined groups of bumblebees. Key words: two-dimensional gas chromatography, Bombus terrestris, cutilar extract

A set of 15 carboxylic acids of various structural types was used to test a new procedure of esterification of these substances in the urine without previous extraction. The acids were first measured by a reference method without the matrix, and then by a procedure, where lyophilized urine was treated with derivatizing reagent alcohol·HCl, which was then evaporated, the residue dissolved in water, esters extracted into an organic solvent and subsequently determined by GC/FID and GC/MS. The work is mainly focused on 2-hydroxyethyl- and carboxymethyl-mercapturic acids, but the procedure was also tested on di-, tri- and hydroxy-carboxylic acids, known as physiological urinary components or markers of metabolic disorders. Key words: carboxylic acids mecapturic acids freeze-dried urine esterification gas chromatography

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,...