Ziel dieser Arbeit war es, neue Ansätze für die Analytik kleiner und hochpolarer Pflanzenschutzmittelrückstände in unterschiedlichsten pflanzlichen Materialien zu entwickeln. Dabei standen die Präzision der Analysenergebnisse, der Arbeitsaufwand, sowie die Robustheit der Methoden und die daraus resultierende Übertragbarkeit für die Routineanalytik im Vordergrund. Hierzu wurden verschiedene Arbeitsschritte von Analysenmethoden an ausgewählten Beispielen von hochpolaren Pflanzenschutzmittelrückständen mit niedrigem Molekulargewicht untersucht.:1 Einleitung 2 Zielstellung 3 Theoretische Grundlagen von Pflanzenschutzmitteln 3.1 Begriffsdefinition 3.2 Forschung und Entwicklung 3.3 Zulassung und rechtliche Grundlagen für Pflanzenschutzmittel 4 Matrixeffekte in der LCMS-Analytik 5 Vorstellung der Zielanalyten und bisherige Analytik 5.1 Triazol-Metaboliten 5.2 Difluoressigsäure 5.3 Ethephon und HEPA 5.4 Phosphonsäure 5.5 Glufosinat-Ammonium und dessen Metaboliten 6 Experimenteller Teil 6.1 Extraktion 6.1.1 Test verschiedener Extraktionsmittel 6.1.2 Test verschiedener Behandlungen während der Extraktion 6.2 Extraktaufreinigung 6.2.1 Extraktverdünnung 6.2.2 Festphasenextraktion 6.2.2.1 Triazol-Metaboliten 6.2.2.1.1 Unpolare SPE 6.2.2.1.2 Kationenaustausch-SPE 6.2.2.1.3 Anionenaustausch-SPE 6.2.2.2 DFA 6.2.2.2.1 Unpolare SPE 6.2.2.2.2 Kationenaustausch-SPE 6.2.2.3 Ethephon und HEPA 6.2.2.3.1 Unpolare SPE 6.2.2.3.2 Kationenaustausch-SPE 6.3 Trennverfahren 6.3.1 Flüssigkeitschromatographie 6.3.2 Kapillarelektrophorese 6.4 Detektion 6.4.1 Tandem-Massenspektrometrie 6.4.2 Differential-Mobilitäts-Spektrometrie 6.4.3 Time-of-Flight Detektion 7 Ergebnisse und Diskussion 7.1 Extraktion 7.1.1 Test verschiedener Extraktionsmittel 7.1.2 Test verschiedener Behandlungen während der Extraktion 7.2 Extraktaufreinigung 7.2.1 Extraktverdünnung 7.2.2 Festphasenextraktion 7.2.2.1 Triazol-Metaboliten 7.2.2.1.1 Unpolare SPE 7.2.2.1.1.1 Vortests 7.2.2.1.1.2 Ergebnisse und Diskussion der finalen Durchführung 7.2.2.1.2 Kationenaustausch-SPE 7.2.2.1.2.1 Wahl des pH-Wertes für die Probenaufgabe auf die SPE 7.2.2.1.2.2 Wahl des pH-Wertes für die Elution vom SPE-Material 7.2.2.1.2.3 Ergebnisse und Diskussion der finalen Durchführung 7.2.2.1.3 Anionenaustausch-SPE 7.2.2.1.3.1 Wahl des pH-Wertes für die Probenaufgabe auf die SPE 7.2.2.1.3.2 Wahl des pH-Wertes für die Elution vom SPE-Material 7.2.2.1.3.3 Ergebnisse und Diskussion der finalen Durchführung 7.2.2.2 DFA 7.2.2.2.1 Unpolare SPE 7.2.2.2.1.1 Vortests 7.2.2.2.1.2 Ergebnis und Diskussion der finalen Durchführung 7.2.2.2.2 Kationenaustausch-SPE 7.2.2.2.3 Anionenaustausch-SPE 7.2.2.2.3.1 Wahl des pH-Wertes für die Probenaufgabe auf die SPE 7.2.2.2.3.2 Wahl des pH-Wertes für die Elution vom SPE-Material 7.2.2.3 Ethephon und HEPA 7.2.2.3.1 Unpolare SPE 7.2.2.3.1.1 Vortests 7.2.2.3.1.2 Ergebnisse und Diskussion der finalen Durchführung 7.2.2.3.2 Kationenaustausch-SPE 7.2.2.3.3 Anionenaustausch-SPE 7.2.2.3.3.1 Wahl des pH-Wertes für die Probenaufgabe auf die SPE 7.2.2.3.3.2 Wahl des pH-Wertes für die Elution vom SPE-Material 7.3 Trennverfahren 7.3.1 Flüssigkeitschromatographie 7.3.1.1 Triazol-Metaboliten 7.3.1.2 DFA 7.3.1.3 Ethephon und HEPA 7.3.1.4 Phosphonsäure 7.3.1.5 Glufosinat und dessen Metaboliten 7.3.2 Kapillarelektrophorese 7.3.2.1 Triazol-Metaboliten 7.3.2.2 Difluoressigsäure 7.3.2.3 Ethephon und HEPA 7.3.2.4 Phosphonsäure 7.4 Detektion 7.4.1 Tandem-Massenspektrometrie 7.4.2 Differential-Mobilitäts-Spektrometrie 7.4.2.1 Triazol-Metaboliten 7.4.2.2 Difluoressigsäure 7.4.3 Time-of-Flight Detektion 7.4.3.1 Triazol-Metaboliten 7.4.3.2 Interferenz des 1,2,4-Triazol 8 Zusammenfassung und Ausblick 9 Anhang 9.1 Verwendete Referenzsubstanzen und Interne Standards 9.2 Physikalisch-chemische Eigenschaften der Analyten 9.3 Strukturformeln der Analyten und Internen Standards 9.4 Anwendungsbeispiele der Analyten 9.5 Massenspektrometrische Parameter der Analyten 9.6 Verwendete LC-Parameter 9.7 Verwendete CE-MS/MS-Parameter 9.8 Verwendete Differential-Mobilitäts-Spektrometrie Parameter 9.9 Verwendete QTOF-Parameter 9.10 Verwendete Formeln 9.11 Ergebnisse 9.11.1 Test verschiedener LC-Phasen 9.11.1.1 Triazol-Metaboliten 9.11.1.1.1 Aquasil C18 9.11.1.1.2 Hypercarb 9.11.1.1.3 Luna SCX 9.11.1.1.4 ZIC-pHILIC 9.11.1.2 Difluoressigsäure 9.11.1.2.1 ZIC-pHILIC 9.11.1.2.2 Hypercarb 9.11.1.3 Ethephon und HEPA 9.11.1.3.1 Hypercarb 9.11.1.3.2 Luna NH2 9.11.1.4 Phosphonsäure 9.11.1.4.1 Hypercarb 9.11.1.5 Glufosinat und dessen Metaboliten 9.11.1.5.1 Hypercarb 9.11.1.5.2 ZIC-pHILIC 9.11.2 Validierung LC-DMS-MS/MS für TDMs 9.11.3 Validierung LC-DMS-MS/MS für DFA 9.11.4 Validierung MRMHR für TDMs 10 Literaturverzeichnis 11 Versicherung und Erklärung gemäß Promotionsordnung

Proteomics is the large-scale study of proteins, particularly their structures and functions. Proteomics has been utilized in medicine for investigation of disease mechanisms and biomarker discovery. Instrumental methods cover sample preparation, protein and peptide separation and mass spectrometry. At present, there is no proteomic method that can be used as universal for every sample. Analytical methods need to be adapted and optimized for certain samples. The aim of this work was to create methodic procedures and to interpret results of experimental and clinical research. The first part of the thesis includes experiments utilizing proteomics to study changes in the plasma proteome clinically relevant porcine model of sepsis-induced peritonitis. Proteomic analyzes were also starting methodological strategies in experiments aimed at kidney physiology and pathophysiology of acute kidney injury during sepsis. Renal biopsies were analyzed in order to study the time course of proteome changes caused by sepsis and surgery. The second part of the thesis contains experiments studying biocompatibility. A method for elution of proteins interacting with adsorbents used in extracorporeal liver support system and with hemodialyzer capillaries was prepared. Analysis of proteins adsorbed to polysulfone...

The goal of my thesis was to find out whether it is possible to distinguish among individual dog breeds from the protein analysis of their fur. This knowledge could be used, for example, in forensic science. In this work, there was also a comparison of the similarity of dog breeds with a wolf, which was domesticated and is considered as an ancestor of the dog. For this study, the hair of three representatives of sixteen dog breeds was collected. To analyse these samples enzyme cleavage was used a trypsin, and mass spectra were obtained by MALDI- TOF MS (Matrix-Assisted Laser Desorption/Ionization - Time of Flight Mass Spectrometry) and LC/MS-MS (Liquid Chromatography with Tandem Mass Spectrometry) methods. The obtained data were evaluated by the PCA (Principal Component Analysis) and LDA (Linear Discriminant Analysis) method. It was found, that individual dog breeds cannot be distinguished using both methods. KEYWORDS proteomics, coat, dog, breeds, mass spectrometry

In recent years, mass spectrometry (MS) become the dominant technology in lipidomic analysis and widely influenced research and diagnosis of diseases of lipid metabolism, e.g. lysosomal storage disorders (LSD) characterized by impairment of the lysosomal functions. Defects in lysosomal processing of sphingolipids SFL belong to the category of sphingolipidoses. This condition has severe and even fatal clinical outcome. The primary aim of this work was to establish quantitative and qualitative methods of SFL analysis useful for research and diagnosis of LSD. At first, semisynthesis of mass labeled lipid standards utilizing immobilized sphingolipid ceramide N-deacylase was performed. Established methods of quantitative analysis were then used to prove the increased excretion of urinary SFL in LSD with characteristic storage in the kidney. Determination of excreted urinary SFL was found useful for differential diagnosis of prosaposin and saposin B deficiences for which routine enzymology is failing. MS also enabled monitoring of individual molecular species (isoforms) of SFL, which led to the finding that their urinary pattern is changing in some LSD. This resulted in the development of new screening method in dry urinary samples based on isoform profile evaluation. Another MS application referred to...

Competitive metallurgical production is almost impossible without the knowledge of the chemical composition of manufactured and finished melt, or the produced product. With growing customer demands for product quality and product characteristics, increasing technology speed and increasing product purity, there is growing demand for analyzes of chemical composition. These demands are very often against each other, especially the increasing demands on the speed and accuracy of analyzes. Analytical requirements may not only apply to the test technique, but also to the quality of the sample, which is inseparable from it. What is recently seen as the biggest drawback, the new technology is trying to use as its main priority. This is undoubtedly true of atomic emission spectrometry.

This diploma thesis researches coffee authenticity problematice, mainly focusing on the authenticity of geographic origin. In the theoretical part of this work, botanical classification is described as well as production technology and processes. The work also includes chemical composition of coffee, describing the major components and changes during production phases. It describes major production areas of the world, in terms of general description and brief history. Problematics with coffee fraud and its identification are also described. Theoretical part also includes general geological description of 17 studied coffee growing regions. Experimental part is devoted to trace amount analysis of selected elements and volatile compounds. The element analysis was conducted using mass spectrometry or optical emission spectrometry, volatile compounds were determined using gas chromatography combined with mass spectrometry detection. Results were statistically described and analyzed, resulting in several discrimination models based on geographic origin.

The diploma thesis is dealing with monitoring of arsenic in foodstuffs. The aim of this thesis is the determination of arsenic in samples of fish and rice and the study of forms, in which arsenic occurs, using speciation analysis. Arsenic is known as a toxic element, but its measure of toxicity depends on the chemical form it occurs in. Arsenic is present in the environment naturally or it gets in the environment by human activities. Complete characterization of arsenic compounds is necessary to understand intake, accumulation, transport, detoxification and activation of this element in the natural environment and living systems. The field of arsenic speciation analysis has grown rapidly in recent years, because determination of the total element content is not sufficient in the case of arsenic. Speciation method was done using separation by high performance liquid chromatography and detection by atomic fluorescence spectrometry with hydride generation. Extractable arsenic was present in the form of nontoxic arsenobetaine in all analysed samples of fish. In samples of rice there was confirmed the presence of toxic inorganic species of arsenic, esspecially As (III), but their concentration was at low level.

Thesis is focused on the application of Raman’s spectometer Ahura First Defender in cooperation with Fire Department. Theoretical part describes principles and development of Raman spectrometry and possibilities of its application. Practical part focus on identification of unknow substances in case studies and interpretation of the results. Final part of thesis evaluates contribution of Raman spectrometry for fast indentification of unknow compounds for Fire Rescue Brigade in the Czech Republic.

This work is focused on the study of penetration of pesticides and pharmaceuticals in the aquatic environment. The aim was to develop and verify the optimal method for long-term monitoring of the occurrence of selected pesticides and pharmaceuticals in surface waters. Two frequently used analgesics ibuprofen and diclofenac, and carbamazepine that is used to treat epilepsy, were chosen from the group of drugs. A group of 41 pesticides were selected for monitoring too. The attention was focused on a group of urea pesticides and phenoxyacetic acids, as well as on selected pesticides falling to the group of priority substances or identified as environmental quality standards. Some of pesticides were selected because their consumption in the Czech Republic exceeds 30 tons a year. Liquid chromatography with tandem mass spectrometry (LC / MS / MS) was used for the determination of selected analytes in surface waters. Two optimized analytical method were developed. Ten analytes were determined in negative ion mode and 34 analytes in positive ion mode. Solid phase extraction (SPE) was used for purification and concentration of the samples. Developed methodology was validated and used for monitoring of selected compounds in samples of surface waters from the Morava River basin. Samples were collected on the rivers Morava and Svratka monthly for two years. Samples from the river Blata and stream Nivnička were collected for one year. Surface water samples were also collected from the river Litava. Four samples were taken throughout the length of the flow during one day. Smaller streams were chosen for monitoring because of the probability of occurrence of pesticides. They flow abundantly agriculturally cultivated areas. Drugs ibuprofen and diclofenac were detected in all taken samples, carbamazepine mostly. Among the most commonly occurring pesticides were bentazon, diuron, isoproturon, carbendazim, MCPA, MCPP, propiconazole, tebuconazole and 2,4-D.

This work is focused on the determination of polar pesticides in surface and waste waters. In this study Mecoprop-P, MCPA, Dichlorprop-P, 2,4-D and MCPBA in differently loaded surface waters and communal waters from sewage treatment plants were analyzed. These herbicides are included in the group identified as environmental quality standards. Some of herbicides (2,4-D and MCPA) were selected because their consumption in the Czech Republic exceeds 30 tons a year. Samples were collected prom three rivers near town of Jaroměř (Elbe, Úpa and Metuje Rivers), from one river near the town of Brno (Svratka River) and from inflow and outflow of two sewage treatment plants (WWTP Jaroměř, WWTP Brno – Modřice). Particular rivers were chosen for monitoring due to the probability of occurrence of herbicides, because they flow through agriculturally cultivated areas. Solid phase extraction (SPE) was used for the isolation of target compounds and their concentration. Gas chromatography with tandem mass spectrometry (GC/MS/MS) was used for the determination of target analytes in surface and waste waters. Selected herbicides were detected in all collected samples.