The ability to effectively analyze particulate matter (PM2.5) in air is becoming increasingly pertinent. Allen Goldstein of the University of California in Berkeley is studying the semi-volatile fraction of organic compounds in PM2.5 through the use of the thermal desorption aerosol gas chromatograph (TAG) system. However, as conventional GC does not provide adequate separation power, the development of comprehensive two-dimensional gas chromatography (GC × GC) was required. GC × GC works more effectively by utilizing a modulator that periodically traps and focuses analytes from a primary column onto a secondary column. This allows for the primary and secondary columns to separate the analytes based on two different properties. This report focuses on the continuing study and enhancement of a modulator designed by Ognjen Panić during his Masters project. Improving and testing the robustness of this dual stage modulator was originally the focus of this project. However, this study led to the development of a single stage modulator. In addition to that, the effect of modulator characteristics such as length of the restriction, total length of the modulator and wall thickness on the modulator performance were studied. A robustness test of the single stage modulator was also completed. Experiments conducted tested the characteristics of the new modulator to ensure it performed effectively and would satisfy the requirements of the TAG system. A study comparing the sensitivity of conventional gas chromatography and GC × GC was also preformed. The sensitivity of GC × GC was on average an order of magnitude better than that of 1D GC.

The ability to effectively analyze particulate matter (PM2.5) in air is becoming increasingly pertinent. Allen Goldstein of the University of California in Berkeley is studying the semi-volatile fraction of organic compounds in PM2.5 through the use of the thermal desorption aerosol gas chromatograph (TAG) system. However, as conventional GC does not provide adequate separation power, the development of comprehensive two-dimensional gas chromatography (GC × GC) was required. GC × GC works more effectively by utilizing a modulator that periodically traps and focuses analytes from a primary column onto a secondary column. This allows for the primary and secondary columns to separate the analytes based on two different properties. This report focuses on the continuing study and enhancement of a modulator designed by Ognjen Panić during his Masters project. Improving and testing the robustness of this dual stage modulator was originally the focus of this project. However, this study led to the development of a single stage modulator. In addition to that, the effect of modulator characteristics such as length of the restriction, total length of the modulator and wall thickness on the modulator performance were studied. A robustness test of the single stage modulator was also completed. Experiments conducted tested the characteristics of the new modulator to ensure it performed effectively and would satisfy the requirements of the TAG system. A study comparing the sensitivity of conventional gas chromatography and GC × GC was also preformed. The sensitivity of GC × GC was on average an order of magnitude better than that of 1D GC.

The biggest limitation to conventional gas chromatography (GC) is limited peak capacity, making the analysis of complex mixtures a difficult or even impossible task. Comprehensive two-dimensional gas chromatography (GC×GC) significantly increases peak capacity and resolution, improves sensitivity and generates structured 3D chromatograms. This is achieved by connecting two columns coated with different stationary phases through a special interface (modulator). The interface samples the first column effluent and periodically injects fractions of this material, as narrow injection pulses, onto the second column for further separation. Commercial instruments achieve this with cryogenic agents. Since this expensive approach permits only in-laboratory analysis, the development of simple, economical and field-capable GC×GC systems is in demand. This report summarizes the fundamentals governing GC×GC separations and a brief history of technological advances in the field. It also documents the construction of a simple interface, devoid of moving parts and cryogenic consumables, and hence highly suitable for field analysis and monitoring applications. Evaluation of the interface suggests on-par performance with more complicated cryogenic modulators. GC×GC separations of technical mixtures of fatty acid methyl esters (FAMEs), common environmental pollutants (EPA 8270), polychlorinated biphenyls (PCBs), pesticides (toxaphene), as well as selected essential oils and major distillation fractions of crude oil indicate very good performance. Most notably, the interface prototype was applied for the first ever time-resolved on-site analysis of the semivolatile organic fraction of urban air particulate matter (PM2.5).

The biggest limitation to conventional gas chromatography (GC) is limited peak capacity, making the analysis of complex mixtures a difficult or even impossible task. Comprehensive two-dimensional gas chromatography (GC×GC) significantly increases peak capacity and resolution, improves sensitivity and generates structured 3D chromatograms. This is achieved by connecting two columns coated with different stationary phases through a special interface (modulator). The interface samples the first column effluent and periodically injects fractions of this material, as narrow injection pulses, onto the second column for further separation. Commercial instruments achieve this with cryogenic agents. Since this expensive approach permits only in-laboratory analysis, the development of simple, economical and field-capable GC×GC systems is in demand. This report summarizes the fundamentals governing GC×GC separations and a brief history of technological advances in the field. It also documents the construction of a simple interface, devoid of moving parts and cryogenic consumables, and hence highly suitable for field analysis and monitoring applications. Evaluation of the interface suggests on-par performance with more complicated cryogenic modulators. GC×GC separations of technical mixtures of fatty acid methyl esters (FAMEs), common environmental pollutants (EPA 8270), polychlorinated biphenyls (PCBs), pesticides (toxaphene), as well as selected essential oils and major distillation fractions of crude oil indicate very good performance. Most notably, the interface prototype was applied for the first ever time-resolved on-site analysis of the semivolatile organic fraction of urban air particulate matter (PM2.5).

Polychlorinated Biphenyls (PCBs) are a group of 209 ‘man-made’ chlorinated organic compounds that were widely used in the 20th century for a variety of industrial uses. PCBs were first commercially produced in the 1929 and were manufactured until the 1980s when their use was phased out due to environmental and human health risks. However, due to their widespread use and persistence they are ubiquitous in the environment and remain a contaminant of concern. The structural properties that determine the persistence of PCBs in humans were therefore elucidated by statistical analysis of data from the National Health and Nutrition Examination Survey (NHANES). PCBs with chlorine bonding in the 2,5-and 2,3,6- positions (and 2- in di- and tri-chlorinated congeners) were rapidly biotransformed and so can be classed as episodic congeners whereas PCBs with chlorine bonding in the 2,3,4-, 2,4,5-, 3,4,5-, and 2,3,4,5- positions were more resistant to biotransformation and can therefore be classed as steady state congeners. A fundamental requirement of using PCB signatures for environmental fingerprinting is an effective analytical method capable of producing high resolution signatures from biological samples. An extraction and clean-up method was developed that was successfully applied to different biological matrices (blood and tissues). A two dimensional gas chromatography with time of flight mass spectrometry (GCxGC-ToFMS) method was designed and optimised to provide a congener specific method capable of identifying 200 out of the 209 PCBs, with detection limits in human serum in the range of 1 to 10 ng g-1 lipid. The extraction and detection methods were used to determine the source of PCB contamination and age date exposure in workers at a transformer dismantling plant. A total of 84 different PCB congeners were identified in the sera of 30 workers with concentrations of the 7 indicator PCBs ranging from 1.2 - 39 μg g-1 lipid. Analysis of PCB signatures was able to distinguish recent from prolonged exposure and also identified an additional source of inhalation exposure in a subgroup of workers. Analysis of 12 different tissue types obtained from the common guillemot (Uria aalge) suggested a high degree of perpetuation between the PCB signature in different tissue types. This shows that comparative assessments can be undertaken between animals using different tissue types and that small (1 g) samples of blood can be used as a non-lethal sampling technique. The regional provenance of 25 wrecked Leach’s storm petrels (Oceanodroma leucorhoa) was also determined using PCB signatures. Results from GCxGC-ToFMS analysis revealed distinctively different PCB signatures in birds from Canada and Europe. The findings reported in this thesis enhance our understanding of PCB signatures in the environment and show how they can be used effectively to age date and identify the source of exposure in humans and animals.

Orientador: Fabio Augusto / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-12T23:52:30Z (GMT). No. of bitstreams: 1 Fidelis_CarlosHenriquedeVasconcelos_D.pdf: 3838436 bytes, checksum: da7eedde4cecf0526ab71e0d126c2a68 (MD5) Previous issue date: 2008 / Resumo: O projeto objetivou estudar os fundamentos que governam um sistema GC×GC, por meio da construção do principal componente de tal sistema, o modulador, sua adaptação a um cromatógrafo a gás comercial e posterior aplicação. O ponto de partida do desenvolvimento instrumental foi a projeção e a construção de um modulador de duplo jato com dois cabeçotes de aço e dióxido de carbono líquido como agente crioscópico. Numa segunda etapa os dois cabeçotes foram substituídos por peça única, utilizando-se ainda o mesmo fluido para resfriamento. Posteriormente o modelo em peça única foi alimentado por nitrogênio líquido. Ambos os sistemas moduladores foram controlados por softwares escritos em laboratório. O primeiro, utilizado em testes iniciais, foi escrito em Visual Basic 3. Em um segundo momento, foi escrito simultaneamente ao desenvolvimento do modulador um software exclusivo para o controle da modulação e registro dos dados. Durante todo o trabalho, algumas aplicações foram feitas para testar o desempenho do sistema. Foram realizadas separações GC×GC principalmente de amostras de gasolina Tipo C. Os resultados obtidos foram extremamente promissores uma vez que ficou demonstrada a aplicabilidade de um sistema modulador para GC×GC utilizandose um forno secundário, externo ao forno do cromatógrafo / Abstract: The aim of this project was to investigate the basic concepts concerning a GC×GC system by adjusting the main system component, the modulator, to a commercial gas chromatograph and its subsequent application. The instrumental development starting point was the projection and construction of a two piece dual jet modulator with carbon dioxide as coolant. In a second stage the two nozzles were replaced by a single piece and the same coolant was used. In sequence nitrogen gas cooled by liquid nitrogen was used as modulating agent with the single piece design. For system control and data acquisition two homemade software were used. The first was written in Visual Basic 3. The second was developed simultaneously to the modulator and is an exclusively applied version. During project development some applications were showed to evaluate system performance. GC×GC separations were performed mainly by using Brazilian gasoline C type. The results obtained are very promising once was demonstrated the applicability of a modulator system using a secondary oven external to the chromatograph oven / Doutorado / Quimica Analitica / Doutor em Ciências

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

No description available.

La chromatographie en phase gazeuse couplée à la spectrométrie de masse simple quadripolaire (GC-qMS) est la technique la plus utilisée dans l’exploration des acides organiques urinaires dans le cadre du diagnostic des aciduries organiques (AO). Cependant, après analyse par GC-MS de plus de 1000 échantillons d'urines recueillis auprès d'enfants atteints de troubles neurologiques, moins de 0,5% d'entre eux ont reçu un diagnostic positif. Ceci témoigne de l’urgence de rechercher de nouveaux biomarqueurs dans le domaine des troubles neurologiques associés à des erreurs innées du métabolisme (EIM).L'objectif principal de cette thèse a été de développer une nouvelle méthode d’analyse des échantillons urinaires par chromatographie bidimensionnelle couplée à la spectrométrie de masse (GCxGC-qMS) et d'évaluer son apport dans le profilage métabolique de ces échantillons. Par ailleurs, dans le cadre du diagnostic des AO, dans un objectif de simplification de la procédure de préparation de l’échantillon, nous avons évalué une autre approche d’analyse directe en temps réel couplée à la spectrométrie de masse « DART-MS » des acides organiques urinaires. La première étape de notre travail a été de développer un protocole de préparation de l’échantillon compatible avec une méthode de séparation bidimensionnelle. Puis, grâce à l’étude systématique des différentes conditions de séparation, nous avons développé une méthode par CGxCG-qMS parfaitement adaptée à la séparation des composés extraits.Après validation analytique, la méthode proposée a été ensuite évaluée par le profilage des composés organiques présents dans des échantillons urinaires prélevés chez des volontaires sains. Par rapport aux profils obtenus par une méthode par GC-qMS classique, optimisée, les profils chromatographiques obtenus par la méthode proposée présentent une sensibilité et une résolution nettement plus élevées. Ceci se traduit par la détection en GCxGC-qMS de nombreux composés supplémentaires, jusqu’à 92, selon les échantillons. Parmi les composés supplémentaires détectés et caractérisés, certains sont d’un intérêt diagnostic reconnu.Malgré les difficultés liées à l’étape de préparation de l’échantillon et au traitement des données, l’ensemble des résultats obtenus par la méthode proposée sur de nombreux échantillons urinaires a confirmé les potentialités et la nécessité d’utiliser la chromatographie bidimensionnelle dans le domaine de la découverte de candidats biomarqueurs. Il s’agit maintenant de poursuivre l’identification des composés supplémentaires détectés dans les échantillons urinaires étudiés et d’appliquer la méthode proposée à un plus grand nombre d’échantillons témoins et pathologiques afin d’essayer d’identifier des candidats biomarqueurs.Enfin, les essais préliminaires par la technique DART-MS, effectués dans un laboratoire de la Direction des Applications Militaires (DAM) du CEA, ont été axés sur la détection d’un panel d’acides organiques représentatifs des différentes anomalies rencontrées lors des AO. Malgré les effets de suppression ionique qui restent à éliminer, les résultats obtenus montrent que cette approche mérite d’être approfondie. / Among inherited metabolic diseases, organic acidemia (OA) or organic aciduria is characterized by urinary excretion of abnormal amounts or types of organic acids. OA is mostly associated with genetic conditions resulting in a specific step of amino acid catabolism dysfunction. Such alterations can produce disease states that range from mild to lethal neurological involvement. Gas chromatography coupled to mass spectrometry (GC-MS) remains the most used analytical technique for detecting specific urinary organic metabolites related to OA. However, after analyzing more than 1,000 urine samples collected from children with neurological disorders, by this technique less than 0.5% of them were positive. Thus, the discovery of new biomarker candidates for other metabolic diseases is urgently needed. The main objective of this thesis was to evaluate a new method using two-dimensional gas chromatography coupled to mass spectrometry (GCxGC-MS) for the both qualitative and quantitative metabolic profiling of children’s urine. As compared to GC-MS, GCxGC-MS shows great resolution power and high peak capacity. For this purpose, we first developed a GCxGC-MS method with an appropriate sample preparation protocol for urinary organic acids profiling. Applied to urine samples of healthy children and children with neurological disorders, the proposed method showed high sensitivity and peak capacity thus opening new possibilities for the characterization of new biomarker candidates. For instance, by this technique, we were able to detect in urines samples more than ninety additional compounds, which are not detected by a conventional GC-MS method. However, taken together the obtained results show that while the GCxGC technique has unmatched power to separate compounds in a complex mixture, the sample preparation protocol remains a limiting step for the precise quantification of the detected compounds. On another hand, in order to reduce the sample preparation step, we evaluated the direct analysis in real time (DART) method for the urinary organic acids screening. The obtained results are very promising.

The use of short columns, higher carrier gas velocity and fast temperature programs in Comprehensive Two-Dimensional Gas Chromatography coupled to Time-of- Flight Mass Spectrometry (GC x GC/TOFMS) technique is expected to increase the speed of analysis up to several orders of magnitude when compared to conventional gas chromatography (GC) or gas chromatography/mass spectrometry (GC/MS). A systematic evaluation of the GC x GC/TOFMS configuration for high-speed applications has received little attention in the literature. The feasibility of High Speed Comprehensive Two-Dimensional Gas Chromatography coupled to Mass Spectrometry (High speed GC x GC/MS) for complex mixtures has been investigated in this thesis. A particular focus was placed on comparing conventional scanning quadrupole mass spectrometry (qMS) with a newly available non-scanning time-of-flight instruments (TOFMS). Experiments were carried out using GC/qMS, GC x GC/qMS, GC/TOFMS and GC x GC/TOFMS both in normal (slow) and fast temperature rates coupled with high frequency modulation in GC x GC. Initially a complex mixture consists of 24 semivolatile compounds was used as the analyte for the above purpose. In the initial experiments parameters like acquisition rate and duty cycle for qMS were determined to evaluate the effectiveness of the instrument for fast analysis. The practical duty cycle value obtained for the qMS was only about 18 % for single ion and one compound at a dwell time of 10 ms in SIM mode. In both high-speed GC/qMS and high-speed GC x GC/qMS techniques only about 40 % of the components in the complex mixture were found to be well separated. The acquisition rate of scanning instruments like qMS is incompatible for fast eluting peaks in high speed GC. TOFMS that has an acquisition rate of several hundred spectra per second offer the potential to define the fast GC peaks accurately. The high quality spectra from TOFMS also enable deconvolution of coeluting peaks in the complex mixtures. The advantage of the automated spectral deconvolution is demonstrated for the identification of the coeluting peaks in the complex mixtures. Coelution of peaks is also observed with highspeed GC/TOFMS technique. The high-speed GC x GC/TOFMS was also tested with two different analyte system ??? A pesticide mixture and platformate (an aromatic mixture) to evaluate the suitability for high-speed analysis of complex mixtures. A poor resolution was observed for the pesticide mixture in the two-dimensional plane and it appeared, as there was nearly no orthogonal separation in the second dimension. The platformate mixture displayed a better two-dimensional separation. Chromatographic peak resolution is not really a primary requirement for locating and identifying the coeluting compounds in high-speed GC x GC/TOFMS technique. However, it was observed that the high-speed GC x GC/TOFMS too faced problem to unscramble the mass spectra of those compounds with similar structure and sharing the same unique masses.