Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Persistent Organic Pollutants (POPs) receive this designation because of the time that remain unchanged in the environment. From the 1960s there was already evidence of the presence of these compounds in sediment, breast milk, fish and eggs. The food chain can be considered a source of contamination of humans by these compounds due to bioaccumulation. The fat contained in milk can accumulate POPs, so this matrix has been used as environmental bioindicator. The currently available methods for the determination of these compounds in breast milk involve, in general, multi-step sample preparation and quantification techniques with low selectivity, have high cost and are time consuming. Thus, this study aimed to develop a fast and efficient method based on sample preparation using the QuEChERS method combined Gas Chromatography coupled to Mass Spectrometry with negative chemical ionization (GC-NCI-MS). In this work we optimized the sample preparation step for the determination of 15 POPs in breast milk (Alpha HCH, beta HCH, gamma HCH, hexachlorobenzene, heptachlor, andrin, dieldrin, endrin, 2,4 DDE, DDT, endosulfan alpha, beta endosulfan, endosulfan sulfate and mirex) using 6 mL of sample extracted with 6 mL of acetonitrile. In the partition step was used 2.6 g MgSO4 and 0.6 g of NaCl. The extract clean-up was performed by dispersive solid-phase extraction of 4 mL of extract and 600 mg of MgSO4 and 200 mg of C18. The extract was evaporated in a Turbovap system and redissolved in 200 μL acetonitrile for the determination by GC-NCI-MS. The method was validated showing linearity for the 15 evaluated compounds. Recoveries of the analytes ranged between 62 and 120% with precision within the acceptable range (RSD <19%). The limits of detection for breast milk, in terms of fat percentage, ranged from 0.75 to 7.5 ng g-1 and the method limits of quantification ranged from 2.5 to 25 ng g-1. The method was applied to 20 samples of breast milk from different regions of the Rio Grande do Sul state, of which only 25% were not contaminated by the studied POPs and the others showed values below the LQ. It is important to emphasize that this is an unpublished work in the area of POPs determination in breast milk by GC-NCI-MS, and could be a basis for future works in this area. / Os Poluentes Orgânicos Persistentes (POPs) recebem essa denominação devido ao tempo que permanecem inalterados no meio ambiente. A partir dos anos 1960 já havia indícios da presença desses compostos em sedimentos, leite materno, peixe e ovos. A cadeia alimentar pode ser considerada uma fonte de contaminação do ser humano por esses compostos devido à bioacumulação. A gordura contida no leite materno pode acumular os POPs, por isso esta matriz vem sendo utilizada como bioindicador ambiental. Os métodos disponíveis atualmente para a determinação desses compostos em leite materno envolvem, em geral, várias etapas de preparo de amostra e quantificação por técnicas pouco seletivas, apresentam alto custo e demanda de tempo. Assim, esse trabalho visou o desenvolvimento de um método rápido e eficiente baseado no preparo de amostra empregando o método QuEChERS combinado a Cromatografia à Gás, acoplada à Espectrometria de Massas com ionização química negativa (GC-NCI-MS). Neste trabalho otimizou-se a etapa de preparo de amostra para a determinação de 15 POPs em leite materno (HCH alfa, HCH beta, HCH gama, hexaclorobenzeno, heptacloro, andrin, dieldrin, endrin, 2,4 DDE, DDT, endosulfan alfa, endosulfan beta, endosulfan sulfato e mirex) utilizando 6 mL de amostra e extração com 6 mL de acetonitrila. Na etapa de partição utilizou-se 2,6 g de MgSO4 e 0,6 g de NaCl. A limpeza do extrato foi realizada por extração em fase sólida dispersiva de 4 mL de extrato e 600 mg de MgSO4 e 200 mg de C18. O extrato foi evaporado em sistema Turbovap e redissolvido em 200 μL de acetonitrila para determinação por GC-(NCI)-MS. O método foi validado apresentando linearidade para os 15 compostos em estudo. As recuperações dos analitos variaram entre 62 e 120%, com valores de precisão dentro da faixa aceitável (RSD < 19%). Os limites de detecção do método para leite materno em termos de percentual de gordura variaram de 0,75 a 7,5 ng g-1e os limites de quantificação do método variaram de 2,5 a 25 ng g-1. O método foi aplicado para 20 amostras de leite materno de diferentes regiões do estado do Rio Grande do Sul, das quais, apenas 25% não apresentaram contaminação pelos POPs estudados e as demais apresentaram valores menores que o LQ. É importante destacar que trata-se de um trabalho inédito na área de determinação de POPs em leite materno empregando GC-NCI-MS, podendo servir de base para trabalhos futuros nessa área.

An efficient two step procedure has been developed using CI GC/MS for analyzing amphetamines and related compounds. The first step allows the analysis of underivatized amphetamines with the necessary sensitivity and specificity to give spectral identification, including differentiation between methamphetamine and phentermine. The second step involves preparing a chiral derivative of the extract to identify d and 1-isomeric composition.

Vernix caseosa is a natural biofilm which covers the skin of a human fetus from the third trimester of pregnancy. It has hydrating, regenerating and anti-microbial effects. The components responsible for these properties of vernix caseosa could be used in the pharmaceutical and cosmetic industries. For this reason, the total composition is analyzed. The lipid components of vernix caseosa consist of squalene, wax esters, sterol esters, 1,2-diol diesters, triacylglycerides, free fatty acids, fatty alcohols, cholesterol, diacylglycerides, monoacylglyceridesů and phospholipids . This study is focused on structure analysis of the 1,2-diol diesters of vernix caseosa. Conditions were optimized for the HPLC separation. Nova -Pak C18 column was used and a gradient of acetonitrile:ethyl acetate was chosen as a mobile phase. Before entering the APCI source ammonium formate was added; ammonium adducts [M +18]+ were formed. Eight scan events was set in MS method. One for the full scan spectrum, second for MS2 spectrum of the precursor ions and six MS3 data-dependent spectra. The position of the double bonds of unsaturated 1,2-diol diesters was determined by fragmentation of their [M+C3H5N]+ adducts. There were identified over 2200 of 1,2-diol diesters differing in lengths of alcohol and fatty acid chains, and...

<p>Aviation fuels are complex mixtures that mostly consist of a wide range of different hydrocarbons but also contain small amounts of heteroatom-containing compounds. Different fuels can have very different physical and chemical properties (e.g., storage and thermal stabilities are influenced by the heteroatom-containing compounds), with some performing better as fuels than others. To address this situation, correlations need to be developed between the chemical composition of fuels and their properties. This requires that the chemical compounds in fuels are correctly characterized, which is challenging. Because of the large number and many different types of organic compounds present in fuels, separation of these compounds by using techniques such as one- (GC) and -two-dimensional gas chromatography (GC×GC) is necessary before mass spectrometric characterization. Furthermore, analysis using traditional electron ionization (EI) mass spectrometry is hampered by excessive fragmentation that often leads to complete absence of a stable molecular radical cation, thus preventing the determination of the molecular weight (MW) of the analyte. To explore alternative methods, GC coupled with methane chemical ionization (CI) triple quadrupole mass spectrometry and GC×GC coupled with methane CI time-of-flight high-resolution mass spectrometry, both in the positive-ion mode, were tested. While both chromatographic techniques separate volatile organic chemicals via boiling point and intermolecular forces, GC×GC methods incorporate a greater level of separation by taking advantage of secondary column of different polarity. This additional level of separation can help separate overlapping compounds that would be impossible in GC. What comes to mass spectrometry analysis, methane CI was found to be more predictable and “softer” than the traditionally employed EI. Several ions revealing the MW of the analyte, e.g., M+•, [M+H-H2]+, and/or [M+H]+, were generated for almost every compound studied with some associated fragmentation. These fragmentation patterns provided invaluable structural information. When combined with the number of carbon atoms in the diagnostic ions, double bond and ring equivalent (DBRE) values, and elemental compositions, all obtained from highly accurate mass measurements performed using the time-of-flight mass spectrometer (but not the quadrupole), the classification of the compounds was possible. In some instances, even the unambiguous identification of individual compounds in aviation fuels was possible.</p>

Les leishmanioses sont des parasitoses en constante évolution, et l’accroissement d’apparitions de résistances vis-à-vis des traitements disponibles en fait une des préoccupations majeures des organismes de santé publique dans le monde. La miltefosine est actuellement le seul antileishmanien actif par voie orale. Son mécanisme d’action implique les lipides et notamment les phospholipides membranaires du parasite.Afin d’évaluer les effets de la miltefosine sur le parasite ainsi que les mécanismes de résistances, une étude lipidomique d’un clone de Leishmania donovani cultivé sous différentes conditions (traité, résistant, résistant-traité) a été réalisée dans le présent travail. Des analyses couplant une séparation des phospholipides en Chromatographie Liquide Haute-Performance à polarité de Phase Normale (NP-HPLC) avec un Spectromètre de Masse (MS) équippé d’une source d’Ionisation ElectroSpray (ESI) ont été traitées par chimiométrie, à l’aide d’une Correction Orthogonale du Signal suivie d’une Analyse Discriminante par Moindre Carrés Partiels (OSC-PLS-DA). Les principales espèces moléculaires permettant de distinguer les différentes cultures ont ensuite fait l’objet d’une identification structurale par spectrométrie de masse en tandem. Des hypothèses métaboliques ont pu être posées.Puis l’étude a été étendue à une plus grande variété de lipides, séparés par NP-HPLC. Pour cela une comparaison des sources d’ionisation à pression atmosphérique (ESI, Ionisation Chimique à Pression Atmosphérique et PhotoIonisation à Pression Atmosphérique) a été nécessaire afin de sélectionner la mieux adaptée pour un tel couplage. Les mécanismes d’action de la miltefosine et de l’amphotéricine B ont alors fait l’objet d’une étude lipidomique. / Leishmaniasis is a more and more spreading disease, and resistance of parasites toward antileishmanial drugs is a concern for public safety organizations troughout the world. Miltefosine is the only oral drug, and its mechanism of action implies membrane lipids, and phospholipids, of parasite cells.In order to assess this mechanism of action, and resistance mechanisms developed, a lipidomic study of Leishmania donovani strains (treated, resistant, treated-resistant) was performed in the present work. A Normal-Phase High-Performance Liquid Chromatography (NP-HPLC) was coupled to an ElectroSpray Ionisation Mass Spectrometer (ESI-MS) to analyze phospholipids, and data were computed using an Orthogonal Signal Correction-Partial Least Squares-Discriminant Analysis (OSC-PLS-DA). Molecular species responsible for the differenciation of strains were then structuraly identified using tandem mass spectrometry. Hypotheses on metabolic pathways implied were then proposed.The study was then extended to a broader range of lipids, also analyzed through NP-HPLC-MS. A comparison of Atmospheric Pressure Ion sources (ESI, Atmospheric Pressure Chemical Ionization and Atmospheric Pressure PhotoIonization) was thus necessary in order to select the most suitable source. A lipidomic study was then performed to assess mechanisms of action and mechanisms of resistance concerning miltefosine and Amphotericin B.

The hydrolysis reaction of N2O5 was investigated at room temperature on two aerosol types using a flow tube coupled to a newly built Chemical Ionization Mass Spectrometer (CIMS). This instrument was fully constructed and optimized during this research period, as well as employed to conduct one of two aerosol studies. The first examined the reaction on ammonium bisulphate aerosols using a new ion detection method, I-•N2O5 cluster formation, which proved to be highly advantageous over the common approach of dissociative charge transfer, yielding a sensitivity for I-•N2O5 of 0.024 Hz/pptv. The uptake coefficients at 30% and 50% relative humidity were 0.0067 ± 0.0002 and 0.0120 ±0.0014, respectively. The second study was performed using a different CIMS previously assembled in the laboratory. In this case, the reaction was investigated on secondary organic aerosols produced through the ozonolysis of α-pinene, and resulted in an uptake coefficient of 8.5x10-5 ± 7x10-6 at 0% relative humidity.

The hydrolysis reaction of N2O5 was investigated at room temperature on two aerosol types using a flow tube coupled to a newly built Chemical Ionization Mass Spectrometer (CIMS). This instrument was fully constructed and optimized during this research period, as well as employed to conduct one of two aerosol studies. The first examined the reaction on ammonium bisulphate aerosols using a new ion detection method, I-•N2O5 cluster formation, which proved to be highly advantageous over the common approach of dissociative charge transfer, yielding a sensitivity for I-•N2O5 of 0.024 Hz/pptv. The uptake coefficients at 30% and 50% relative humidity were 0.0067 ± 0.0002 and 0.0120 ±0.0014, respectively. The second study was performed using a different CIMS previously assembled in the laboratory. In this case, the reaction was investigated on secondary organic aerosols produced through the ozonolysis of α-pinene, and resulted in an uptake coefficient of 8.5x10-5 ± 7x10-6 at 0% relative humidity.

The goal of this project was to develop a rapid separation and detection method for analyzing organic compounds in smokeless powders and then test its applicability on gunshot residue (GSR) samples. In this project, a total of 20 common smokeless powder additives and their decomposition products were separated by ultra performance liquid chromatography (UPLC) and confirmed by tandem mass spectrometry (MS/MS) using multiple reaction monitoring mode (MRM). Some of the targeted compounds included diphenylamines, centralites, nitrotoluenes, nitroglycerin, and various phthalates. The compounds were ionized in the MS source using simultaneous positive and negative electrospray ionization (ESI) with negative atmospheric pressure chemical ionization (APCI) in order to detect all compounds in a single analysis. The developed UPLC/MS/MS method was applied to commercially available smokeless powders and gunshot residue samples recovered from the hands of shooters, spent cartridges, and smokeless powder retrieved from unfired cartridges. Distinct compositions were identified for smokeless powders from different manufacturers and from separate manufacturing lots. The procedure also produced specific chemical profiles when tested on gunshot residues from different manufacturers. Overall, this thesis represents the development of a rapid and reproducible procedure capable of simultaneously detecting the widest possible range of components present in organic gunshot residue.

No description available.

The work described herein is related to gas-phase ion-molecule reactions studied by using mass spectrometry. Chapter 2 describes density functional theory, a method used in chapters 4 and 5 to propose reaction mechanisms for reactions previously observed by others by using mass spectrometry. Chapter 3 describes a study that demonstrates that the fragmentation of saturated hydrocarbons occurs due to proton-transfer reactions that occur between these species and protonated molecules generated from molecules present in air such as nitrogen and water. Saturated hydrocarbons are studied in a wide variety of fields, and better methods to analyze complex mixtures of these compounds would facilitate their analysis. Chapter 4 discusses mechanisms of reactions for previously studied ion-molecule reactions of pyridynes studied by others by using mass spectrometry. Reactions of pyridynes are important to study arynes have been previously used in organic synthesis, and pyridine moieties are particularly common in biological compounds. Chapter 5 discusses density functional theory calculations used to determine why some organic polyradical undergo hydride abstractions from cyclohexane while others do not. The study discusses reactions taking place between both singlet and triplets states of the 2,5-didehydropyridinium cation and cyclohexane as a model, which are compared to reactions of the 2-pyridyl cation and 2-dehydropyridinium cation with cyclohexane. These studies may help improve our understanding of the reactivity-controlling factors of organic polyradicals, which may help improve toxic drug candidates like cytostatic enediynes.