Rozprawy doktorskie na temat „NMR spectroscopic techniques”

Aerosols are ubiquitous in the atmosphere, and play a key role in several environmental processes at local and global scale affecting visibility, air quality, and the climate system. In the present study, proton nuclear magnetic resonance (H-NMR) spectroscopy and aerosol mass spectrometry implemented by an “aerosol chemical speciation monitor” (ACMS) were employed to investigate the chemical composition of submicrometric aerosol collected during a series of intensive field campaigns and to resolve its composition into source-specific fractions including distinct types of primary and secondary organic aerosols (POA and SOA). During the first study, aerosol samples were collected in the Po Valley, in Bologna (urban site) and San Pietro Capofiume (rural site), during the summer 2012 (PEGASOS project) and fall, winter seasons 2013 (SUPERSITO campaigns). The results show that biomass burning POA and SOA dominate the composition of oxygenated organic aerosols in the Po Valley in the cold season, whereas humic-like substances (HULIS), related to chemically-aged SOA types, are prevalent in the summertime. In the second part of this thesis work, SOA samples were produced in laboratory using a flow tube reactor designed to simulate prolonged photochemical ageing. The samples were analyzed by H-NMR and high-liquid performance chromatography (HPLC) for the characterization of isoprene, α-pinene and naphthalene SOA products. The third part of this work was dedicated to two field campaigns at coastal and marine sites in the central Mediterranean region: the first one in Civitavecchia – the port of Rome, for the AIR-SEA Lab project -, and another one onboard the Amerigo Vespucci vessel during the project Dual Use Vespucci. Results showed that PM1 total concentration resulted only 22% lower over the open sea with respect to ports, but the fossil fuel component of OA (the hydrocarbon fraction) was much more enriched in harbors than in off-shore areas.

Flap endonuclease 1 (FEN1) is a member of a 5’ nuclease superfamily involved in DNA replication and repair. FEN1 hydrolyses the phosphodiester bond one nucleotide into the duplex region of bifurcated double-flapped structures as found in lagging-strand DNA synthesis. These flap structures need to be cut in a very specific manner on the order of around 106 times per cell cycle. Therefore FEN1 is seen as an essential enzyme that maintains genomic integrity across all life forms. How FEN1 achieves its molecular recognition for a chemically very similar but structurally different DNA substrate and how it achieves catalysis on a biochemically relevant timescale are key questions to understand the protein system. This thesis describes some of the mechanistic studies used to understand how the structure and dynamics of hFEN1 relate to its function. It was proposed that T5 bacteriophage FEN was a catalytically perfect, or diffusion-limited enzyme, yet its main rate-limiting step after substrate binding was non-chemistry related. To ascertain whether this was true for hFEN1, the effect of leaving group pKa using 2’ modified double flapped substrates on rates of catalysis was measured. It was found that both apparent second order rates and first order single turnover rates of catalysis were insensitive to leaving group pKa. Furthermore by supplementing the reaction with glycerol, an unexpectedly high viscosity dependence was observed. The explanation for this is likely the presence of another physical step in the catalytic cycle affected by viscosity. Previous structural and biophysical studies of hFEN1 identified a helical arch, which was thought to be disordered as it could accommodate bulky 5’ flaps through it. Furthermore, the arch is key for positioning the 5’ flap into the active site. Using NMR spectroscopic techniques the solution state conformation of apo-hFEN1 was analysed. The arch was found to be disordered, but the C- terminal portion of it was transiently sampling α-helical φ,ψ space, while the other half was in an extended conformation. Another DNA recognition region, the α2-α3 loop was also found to be disordered. Various ligands and substrates were found to alter the structure and the dynamics of hFEN1. Addition of substrate DNA slowed the motion of the arch and α2-α3 loop to a millisecond timescale. Equally addition of a single monophosphate nucleotide had an effect on the dynamics of the top of the arch, despite binding in the active site. Furthermore, titration of calcium ions into the active site when DNA was present on the enzyme resulted in large perturbations to substrate recognition sites distant from the active site. This potentially links the specificity of these regions to activity within the active site.

The effects of cardioplegic and reperfusion conditions on energy metabolites and cardiac function were investigated in order to define better conditions for heart preservation (4 or 8 hrs). Myocardial energy metabolites (ATP, PCr and inorganic phosphate), intracellular pH and contractile function were followed using $\sp{31}$P NMR spectroscopy and left intra-ventricular balloon, respectively, during preservation and reperfusion in isolated pig and rat hearts. These hearts were subjected to various conditions of cardioplegia and reperfusion which involved re-arrest perfusion following ischemic preservation, increased concentrations of buffer and Mg$\sp{++}$ in cardioplegic and reperfusion solutions, and an intracellular-type cardioplegic solution. The effect of re-arrest perfusion was tested by comparing the recovery of energy metabolites and contractile function between pig hearts subjected to a secondary cardioplegic solution (S-C-S) prior to Krebs-Henseleit (K-H) solution and those reperfused with K-H solution alone after 8 hours of ischemic preservation at 12$\sp\circ$C. The levels of ATP and PCr during reperfusion in both groups of hearts were comparable whereas the left ventricular developed pressure was significantly higher in the hearts reperfused with S-C-S than in those reperfused only with K-H solution. The reperfusion-induced ventricular fibrillation that occurred in K-H reperfused hearts was prevented by re-arrest perfusion. A cardioplegic solution containing 150 mmol/L MOPS (higher buffer cardioplegic solution) helped to maintain intracellular pH during 8 hours of ischemic preservation. However, it did not affect the levels of energy metabolites during preservation and contractile function during reperfusion. The effects of 16 mmol/L Mg$\sp{++}$ in cardioplegic and reperfusion solution were evaluated using both rat and pig hearts. In working rat hearts, 16 mmol/L Mg$\sp{++}$ in cardioplegic solution did not alter myocardial oxygen consumption and contractile function following 30 minutes of normothermic preservation. Moreover, Langendorff pig hearts preserved with either 0 or 16 mmol/L Mg$\sp{++}$ cardioplegic solution showed similar decrements in energy metabolites during 4 hours of ischemic preservation at 12$\sp\circ$C and recovery of contractile function during reperfusion while 16 mmol/L Mg$\sp{++}$ in K-H solution resulted in a dramatic decline of contractile function. Furthermore, 16 mmol/L Mg$\sp{++}$ in S-C-S also did not affect the levels of high energy phosphates and contractile function during reperfusion. At 4$\sp\circ$C the pig hearts stored with either University of Wisconsin solution (UW, an intracellular-type cardioplegic solution) or St Thomas' solution (an extracellular-type cardioplegic solution) showed comparable changes in energy metabolites during 8 hours of preservation and a similar recovery of contractile function during reperfusion. However, at 12$\sp\circ$C, hearts stored in UW solution showed rapid decrease in ATP and PCr during preservation and significantly poorer functional recovery during reperfusion; four of eight hearts stored in UW solution at 12$\sp\circ$C showed the "stone heart" phenomenon with disappearance of PCr and ATP upon reperfusion. The addition of 0.5 mmol/L Ca$\sp{++}$ to UW solution significantly improved contractile function and prevented the occurrence of the "stone heart" phenomenon with stable levels of high energy phosphates. (Abstract shortened by UMI.)

This work focuses on pigmy sperm whale and horse myoglobins (Mbs), which are distinguished by a single heme pocket residue variant in the CD3 position, when the heme iron is in the +3 oxidation state (i.e. the met form). The strategy employed is as follows: (i) assign heme peripheral protons; (ii) assign the amino acid residues from the heme cavity; (iii) assess the dynamics of ligand binding in the active site by means of hydrogen Iability, solvent isotope effects, and heme-insertion isomer trapping, all by NMR methods. The results of these studies portray dynamic solution structure of the Mb ligand binding site, and provide a set of standard parameters for the studies of larger hemoproteins. The findings are also important for understanding protein-ligand interactions in general. My research investigates the mixed spin metazido and metimidazole complexes of Mbs for the following reasons. First, the allosteric properties of hemoglobin arise mainly from the transition between its two possible quaternary structures. This can be studied by paramagnetic NMR because it is one of the most sensitive tools in terms of changes in the molecular and/or electronic structure of the heme. Second, both the N₃- and imidazole (lm-) complexes are good compromises, in terms of sizes, between the small diatomic oxygen or CN⁻ molecules and the bulky phenyl group. Thus, we can determine the influence of ligand size on structural perturbation of the Heme crevice by comparison among the different size groups. Third, the saturation-transfer phenomenon between metMbIm and metMbH₂0 provides a route to assignments in metMbH₂0 by using assignments of metMbIm. This is crucial because metMbH₂0 is the basis of theoretical calculations of the isotropic shift due to axial ligand field in pure high-spin hemoproteins. Finally, the importance of the metMbIm is underscored by the fact that it is a bis-imidazolium complex, which can then serve as a model other bis-histidyl proteins. Most of the heme peripheral resonances of metEqMbIm and metEqMbN₃ were identified by means of two-dimensional NOESY,COSY, and EXSY spectroscopy. The strongly relaxed upfield protons in metMbIm were assigned based on steady-state 1D NOE and T₁ experiments. Based on the results from metMblm in which saturation transfer of one upfield resonance led to two different free ligand peaks, bound Im equilibration was envisioned and proven by the divergence of broad downfield heme methyl peaks into two peaks each, showing distinctive population preference of each isomer. Dicyanoheme probe, as well as hydrogen Iability comparison studies between pigmy sperm whale Mb and horse Mb in the azido and imidazole states, asserts that single variant pocket residue CD3 is crucial in gating the ligand mobility into and out of the active site. The assignments of heme peripheral and upfield resonances enabled the subsequent assignments of some heme pocket amino acid residues. The facile exchange of bound Im with solvent H₂0 lays the ground work for identification of heme pocket residues in metMbH₂0. Furthermore, while deuterated heme previously allowed only assignment of the non-diastereomeric specific heme 2-vinyl β proton, saturation-transfer from horse imidazole Mb affords the specific identification of 2Hᵦt.

Spectroscopic techniques allow for studies of materials and organisms on the atomic and molecular level. Examples of such techniques are nuclear magnetic resonance (NMR) spectroscopy—one of the principal techniques to obtain physical, chemical, electronic and structural information about molecules—and magnetic resonance imaging (MRI)—an important medical imaging technique for, e.g., visualization of the internal structure of the human body. The less well-known spectroscopic technique of nuclear quadrupole resonance (NQR) is related to NMR and MRI but with the difference that no external magnetic field is needed. NQR has found applications in, e.g., detection of explosives and narcotics. The first part of this thesis is focused on detection and identification of solid and liquid explosives using both NQR and NMR data. Methods allowing for uncertainties in the assumed signal amplitudes are proposed, as well as methods for estimation of model parameters that allow for non-uniform sampling of the data. The second part treats two medical applications. Firstly, new, fast methods for parameter estimation in MRI data are presented. MRI can be used for, e.g., the diagnosis of anomalies in the skin or in the brain. The presented methods allow for a significant decrease in computational complexity without loss in performance. Secondly, the estimation of blood flow velo-city using medical ultrasound scanners is addressed. Information about anomalies in the blood flow dynamics is an important tool for the diagnosis of, for example, stenosis and atherosclerosis. The presented methods make no assumption on the sampling schemes, allowing for duplex mode transmissions where B-mode images are interleaved with the Doppler emissions.

The analysis of mixtures is a subject that spans several different analytical techniques. In NMR, a powerful technique for this analysis is Diffusion-Ordered NMR Spectroscopy (DOSY), in which signals from different chemical species can be distinguished by their different diffusion coefficients. DOSY NMR allows an analysis of mixture components and their interactions in a non-invasive way and is proving to be an accurate and time effective method for looking at mixtures.An in-depth analysis of DOSY NMR is presented using the commercial mixture “monoacetin”. The chemically cognate species in this mixture produce complex and overlapping multiplet signals in 1H NMR that are difficult to assign and interpret. A previous analysis of this mixture used 1H NMR together with Liquid Chromatography (LC) and Gas Chromatography (GC) to identify the components present, but failed to provide complete assignments of all the signals. Here, the possibility of using DOSY as an alternative to hyphenated techniques is examined, and it is shown that a full analysis of the spectrum of “monoacetin” is possible with careful selection of experimental parameters and processing techniques, without recourse to chromatography. DOSY NMR can be ineffective when signals overlap and/or diffusion coefficients are similar. Many methods have been proposed to overcome these problems, and some of these are presented here. In order to increase resolving power, it is possible to gather further information about a mixture and incorporate this into diffusion experiments as another dimension. This creates a 3D dataset that can be analysed using a multiway method, such as PARAFAC, to extract the component spectra. This method is explored for the mixture “monoacetin” that has been partially separated by high-performance liquid chromatography. Resolution of two out of four components was achieved from poor HPLC separation; the decomposition obtained the component spectra, diffusional decay and HPLC elution profile for these components. Improved HPLC separation should result in further resolution.Diffusion coefficients of different mixture species can be manipulated by changing the matrix in which they diffuse: Matrix-Assisted DOSY (MAD). Previous techniques have involved either improving resolution in the diffusion domain or aiming to improve resolution in chemical shift. A method is presented here that simultaneously addresses both problems in a chemically-selective way, using lanthanide shift reagents. The chemically-selective binding of the LSR to mixture components can both enhance chemical shift dispersion and increase diffusion resolution in DOSY. This neatly deals with the two main drawbacks of the DOSY experiment, and is demonstrated using a mixture of an alkane, alcohol and aldehyde. The manipulation of a molecule’s electrostatic charge through pH control has been investigated, where small ions with a high charge density would be highly solvated, resulting in a change in D. The effect, however, was not measurable and so the indirect effect of pH on the interaction of charged species with the cationic micelle CTAB is presented, where an increase in resolution between of mixture of aspirin and salicylic acid is achieved.Although DOSY NMR is a powerful tool for mixture analysis, in recent years it has been used for studying molecular interactions. An example of this is presented here where species aggregate under specific conditions, a process identified by DOSY NMR.

The aim of this work was to assess the suitability of vibrational spectroscopic techniques (Raman, FT-IR and FT-NIR spectroscopy) as a means for the solid-state structural analysis of pharmaceuticals. Budesonide, fluticasone propionate, salbutamol hemisulfate, terbutaline hemisulfate, ipratropium bromide, polymorphic forms of salmeterol xinafoate and two polymorphic forms of sulfathiazole were selected since they are used in the management of certain respiratory disorders and from different chemical and pharmacological entities along with some pharmaceutical excipients. Conventional visual examination is not sufficient to identify and differentiate spectra between different pharmaceuticals. To confirm the assignment of key molecular vibrational band signatures, quantum chemical calculations of the vibrational spectra were employed for better understanding of the first five selected drugs. The nondestructive nature of the vibrational spectroscopic techniques and the success of quantum chemical calculations demonstrated in this work have indeed offered a new dimension for the rapid identification and characterisation of pharmaceuticals and essentially warrant further research. The application of simultaneous in situ Raman spectroscopy and differential scanning calorimetry for the preliminary investigation of the polymorphic transformation of salmeterol xinafoate polymorphs and two polymorphic forms of sulfathiazole has also been explored in this work leading to the development of a new method for the solid-state estimation of the transition temperature of entantiotropically related pharmaceutical polymorphs which represents the first analytical record of the use of this approach for pharmaceutical polymorphs.

La maladie de Parkinson est une maladie neurodégénérative du système cerveau central conduisant à l'apparition des troubles moteurs caractéristiques de la maladie : akinésie, rigidité et tremblement de repos. La perte des neurones dopaminergiques de la voie nigro-striée va conduire à des modifications biochimiques au niveau du putamen. Notamment, les travaux réalisés en électrophysiologie, microdialyse et spectroscopie par résonance magnétique (SRM) suggèrent une hyperactivité de la voie glutamatergique cortico-striatale associée à un changement du microenvironnement glial au niveau du putamen. Ces observations conduisent à penser à une adaptation du cycle glutamate-glutamine ayant lieu entre les neurones et les astrocytes en réponse à la perte neuronale. Ainsi, dans ce travail de thèse, deux approches ont été développées afin de suivre par SRM les changement métaboliques impliqués dans la pathologie parkinsonienne, notamment les variations des concentrations en glutamate et glutamine dans le putamen. Une première approche de la quantification des métabolites cérébraux par spectroscopie du 1H, technique couramment utilisée en clinique, a été utilisée pour suivre l'évolution des métabolites d'intérêt chez des patients parkinsoniens à jeun ou suite à la prise d'un traitement dopaminergique. Si cette étude a révélé des changements de concentration en N-acetylaspartate, créatine et myoinositol chez les patiens parkinsoniens, aucun changement du métabolisme glutamatergique n'a pu être observé par cette technique, peut-être à cause d'un manque de sensibilité de la technique pour discriminer les pools de glutamate et de glutamine. De ce fait, une nouvelle approche de SRM du Carbone 13C a été développée pour le suivi du cycle glutamate-glutamine in vivo, c'est la polarisation dynamique nucléaire (PDN). Grâce à la haute sensibilité de cette technique, il est désormais possible de suivre des voies métaboliques in vivo en temps réel. La mise en place et l'optimisation de la PDN pour le suivi du cycle glutamate-glutamine a été un des objectifs au cours de ce projet de thèse. Validée sur un groupe d'animaux contrôle, cette technique offre un avenir prometteur pour l'analyse de ce flux dans les pathologies neurodégénératives. En conclusion, les stratégies diagnostiques en clinique par SRM du 1H restent, à l'heure actuelle, peu sensibles pour l'étude des modifications du cycle glutamate-glutamine in vivo chez l'homme. Les développements technologiques réalisés au cours de ce travail de thèse notamment avec la PDN du 13C laissent entrevoir une nouvelle approche pour le suivi en temps réel de ce métabolisme cérébral. Si la PDN est principalement utilisée dans des études précliniques, la disponibilité de nouveaux systèmes cliniques pourrait permettre son avènement en tant que nouvelle stratégie de diagnostic en imagerie clinique
Parkinson's disease is a neurodegenerative disorder characterized by motor troubles such as akinesia, rigidity and tremor. The loss of dopaminergic neurons from the nigro-striatal pathway will lead to biochemical changes in the putamen. Especially, works on electrophysiology, micro dialysis and magnetic resonance spectroscopy (MRS) suggests hyperactivity of the glutamatergic cortico-striatal pathway associated with glial microenvironment changes. These observations suggest a modification of the glutamate-glutamine cycle occurring between neurons and astrocytes in response to neuronal loss.In this thesis, two approaches have been developed in order to follow by MRS the metabolic changes occuring in Parkinson's disease. In particular, we want to follow the changes in glutamate-glutamine cycle inside the putamen.in a first study, a a 1H MRS approach was used to assess the metabolic changes inside the putamen of Parkinson's disease without or under dopaminergic treatment. In this study, changes in N-acetylaspartate, creatine and myo-inositol were observed in Parkinsonian patients, but no change in glutamatergic metabolism was observed. This could be due to the lack of sensitivity of the technique to differentiate glutamate and glutamine pools.Thus, we chose to use a new 13C carbon MRS approach in order to follow dynamically in vivo the glutamate-glutamine cycle inside the brain: dynamic nuclear polarization (DNP). Thanks to the high sensitivity of this technique, it is now possible to follow metabolic pathways in vivo in real time. The implementation of DNP was assessed under a control group of animals. This technique offers a new promising tool for the analysis of this flow under pathologic conditions.To conclude, the MRS strategies for clinical diagnostic strategies remain, at present, poorly sensitive for the study of glutamate-glutamine cycle in vivo in humans. The development of DNP opens the door to a new approach for real-time monitoring of this cerebral metabolism Even if DNP is mainly used in preclinical studies at present, the development of new clinical systems could lead to its emergence as a new diagnostic strategy in clinical imaging

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Includes bibliographical references.
The need to come up with new and novel antibiotics that utilize unique mechanisms, to which bacteria cannot generate resistance, was the main motivation of this study. Tyrocidine peptides are non-selective antibiotics that have such properties. However, very limited information is available about their mechanism of action. The aim of this study was to determine the mechanism of action of tyrocidine peptides, tyrocidine A, tyrocidine B and tyrocidine C.

In the past years, metabolomics has progressed greatly, providing a reliable and high-throughput approach particularly feasible for the study of complex biological systems. Indeed, thanks to the development of powerful analytical methods capable of screening a large number of chemical compounds in a sample, nowadays metabolomics plays an important role in gaining biological insights toward the influence of internal (genetic and developmental) and external (environmental) factors on phenotypes. The work presented in this Ph.D. thesis shows examples of applications of NMR-based metabolomics to the study of bivalves, facing challenges of interest in both food and environmental sciences. The outcome of such studies yield insights, at molecular level, into several aspects concerning the impact of different storage conditions on shellfish quality and the effect of natural and anthropogenic environmental stressors on bivalves’ metabolic profiles. Firstly, the effect of different cold storage conditions on the hydrosoluble chemical components of Mytilus galloprovincialis (Lamarck, 1819) was investigated for the first time by NMR-based metabolomics. The data revealed substantial time-related changes in the metabolic profiles of mussels stored at 0 °C and 4 °C. The observed biochemical modifications were in good agreement with the microbiological quality of samples, reflecting changes in their microbial loads. These results confirmed the potential use of metabolomics as a reliable method to assess seafood freshness. Secondly, a metabolomic approach was also applied to study the effect of short-term exposure to heavy metals on two different clams’ species: Ruditapes decussatus (Linnaeus, 1758) and Ruditapes philippinarum (Adams & Reeve, 1850). Heavy metals are considered to be among the most harmful pollutants that can contaminate marine environments. The toxicity of trace metals gives rise from their persistent nature which lead to environmental accumulation. Given the high accumulation rates associated with their filter feeding attitude, bivalves mollusks are considered to be feasible monitoring organisms and are widely used in biomonitoring programs. R. decussatus and R. philippinarum, two bivalves’ species widely distributed along the Italian coasts, were selected in this Ph.D. project for assessing lead and zinc effects on their metabolic profiles. The results evidenced a sensible short-time metabolic response upon metal exposure, pointing out a main variability in the content of amino acids and organic osmolytes in relation to both metal nature and bivalve species. These findings show that NMR-based metabolomics has the required sensitivity and specificity to gain insights into the biochemical consequences arising upon heavy metals exposure, providing thus a useful tool for the identification of putative biomarkers as fast and sensitive indicators of contaminant-induced stress. Finally, 1H NMR-based metabolomics was applied with the aim to assess the effects of seasonal change on Ruditapes decussatus metabolic profile. The results of the present study demonstrated that the combined use of advanced multivariate statistical techniques with NMR spectroscopy is a feasible approach to discriminate specimens of R. decussatus according to the sampling season. Moreover, the sensitivity of this analytical tool allowed the individuation of those metabolites whose relative amount significantly varied according to seasonal change (alanine and glycine), paving the way for further investigations that would contribute to achieve additional insights on bivalves’ bio-ecological framework.

The global progression towards sustainable energy, materials and chemicals requires novel and improved analytical tools to understand and optimize lignocellulosic biomass utilization. In an effort to advance lignocellulose characterization, gain insights into biomass processing, and obtain novel perspectives on cell wall ultrastructure, this study utilizes three principal polymer characterization techniques, namely compressive-torsion dynamic mechanical analysis (DMA), deuterium quadrupolar nuclear magnetic resonance (2H NMR) and rheo-infrared spectroscopy. A novel parallel-plate compressive-torsion DMA protocol is developed to analyze very small solvent-plasticized biomass specimens with or without mechanical integrity. The benefits and limitations of this technique are demonstrated by comparing it to a conventional tensile-torsion DMA while analyzing various solvent-plasticized lignocelluloses. The rheology of wood in various organic solvents is studied through dynamic thermal scans, Time/temperature superposition (TTS) and fragility analysis. Plasticizing solvents and wood grain orientation significantly affected the lignin glass-transition temperature. Dynamic TTS reveals that while all storage modulus data shift smoothly, the thermorheological complexity of solvent-plasticized wood becomes evident in loss component master curves. It is argued that the plasticized lignocellulose TTS is insightful and potentially useful, although it fails to satisfy the classic TTS validity criteria. Subsequently, it is justified that the fragility analysis is a better suited treatment than the WLF model to investigate cooperative segmental motions of plasticized wood. Deuterium quadrupolar NMR reveals a new perspective on the orientation of amorphous wood polymers and two distinct amorphous polymer domains: a highly oriented phase in the S2 layer of the secondary cell wall and an isotropic phase postulated to occur in the compound middle lamella (CML). If the origin of the isotropic phase is confirmed to arise from the CML, then this technique provides a way to independently investigate the morphology and phase dynamics of CML and S2 in an intact tissue, and should bring novel insights into deconstructive strategies specific to the oriented and unoriented domains. Finally the effects of a wood-adhesion promoter (hydroxymethyl resorcinol, HMR) on in-situ wood polymers are studied to elucidate the still unresolved HMR-lignocellulose interactions. DMA, creep-TTS and 2H NMR reveal that HMR increases the crosslink density and restricts the mobility of wood amorphous phase. Rheo-IR spectroscopy shows that the molecular stress-transfer mechanism is altered within the wood cell wall.
Ph. D.

La presente tesis doctoral se centra en el estudio de técnicas de mejora de resolución de la espectroscopía por Resonancia Magnética Nuclear (RMN) para una más simple y precisa elucidación estructural de moléculas pequeñas. El trabajo se ha presentado como un compendio de siete publicaciones escritas para diversas prestigiosas revistas científicas. Las publicaciones desarrollan en profundidad la aplicación de las mejoras de resolución para i) una medida precisa y eficiente de las constantes de acoplamiento tanto homo- como heteronucleares e ii) la optimización de las medidas de parámetros anisotrópicos a través de medios débilmente alineados. Estos dos tópicos se desarrollan mediante: 1. La aplicación de modernas técnicas de RMN para la mejora de laresolución, tanto digital como de señal, en experimentos 2D sin comprometer el tiempo experimental. 2. El diseño de nuevas secuencias de pulsos para la medida de conectividades de largo alcance heteronuclear. 3. La implementación de secuencias de pulsos ya existentes y el diseño de nuevas para el estudio de muestra en medio aniostrópicos. 4. El estudio de nuevos métodos en RMN para la discriminación mediante parámetros anisotrópicos. 5. La aplicación de protocolos para la automatización y simplificación de las medidas de contantes de acomplamientos en medio isotrópicos y anisotrópicos.
The present doctoral thesis is focused on the study of resolution-enhanced techniques for Nuclear Magnetic Resonance (NMR) spectroscopy in order to achieve a simpler and more precise structural elucidation of small molecules. The work has been presented as a compendium of seven publications written in several prestigious scientific journals. The publications develop in depth the application of resolution improvements for i) a precise and efficient measurement of homo- and heteronuclear coupling constants and ii) an optimization of anisotropic parameter measurements through weakly aligned means. These two topics are developed by: 1. The practical implementation of modern NMR techniques to improve the digital and signal resolution of 2D experiments without compromising the experimental time. 2. The design of improved pulse sequences to achieve the measurement of longer- range heteronuclear connectivities. 3. Implementation of novel NMR approaches to study samples in anisotropic environments, as well as the design of new pulse sequences adapted to the experimental measurement of NMR parameters in these media. 4. Evaluation of new NMR strategies for efficient structure discrimination using anisotropic NMR parameters. 5. Application of protocols for the easy automatization and measurement of coupling constants in isotropic and anisotropic media.

Consumers and food authorities are, to an increasing extent, concerned about factors such as the origin of food, how it is produced, and if it is healthy and safe. There are methods for general quality control to map the safety and nutritional value; however there is a need for suitable analytical methods to verify information such as the production method (wild/farmed), geographical origin, species, and process history of foods. This thesis evaluates the applicability of using nuclear magnetic resonance (NMR) spectroscopy combined with pattern recognition techniques for authentication of foodstuffs. Fish and marine oils were chosen as materials. 13C NMR was applied to authenticate marine oils and muscle lipids of both fatty and lean fish, according to production method (wild/farmed), geographical origin, species, and process history. 1H NMR was applied on low molecular weight compounds extracted from cod muscle to authenticate fish according to species and processing conditions. 13C NMR combined with pattern recognition techniques enabled the differentiation of marine oils according to wild/farmed and geographical origin of the raw material. It is suggested that this was mainly due to the different diets of the fish from which the oil was produced. It was also possible to authenticate marine oils according to species, and to say something about the level of mixtures detectable. The sn-2 position specificity of fatty acids in triacylglycerols was shown to be an important characteristic to separate oils of different species. Esterified fish oil (concentrates) could easily be differentiated from natural fish oil by their 13C NMR profile. 13C NMR on muscle lipids, combined with pattern recognition techniques enabled the classification of wild and farmed salmon. The classification according to geographical origin was somewhat more complicated. A combination of analytical methods may be the best approach to obtain reliable results on geographical origin of fish. The analysis of lean fish showed that it was possible to classify lean gadoids according to species, and two stocks of Atlantic cod could be differentiated. There were also minor differences in the sn-2 position specificity of 22:6n-3 in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) among the species investigated. 1H NMR on water soluble extracts of fish muscle provided information about a wide range of compounds, and the two species investigated (cod and haddock) displayed different 1HNMR profiles. Dimethylamine (DMA) was used as a marker for frozen, non processed fish. When applying the 1H NMR data in pattern recognition techniques, frozen fish could be differentiated from non-frozen fish, and in the classification of the cod of the different processing methods 80% of the samples were correctly classified. Common for the methods presented in this thesis, NMR spectroscopy combined with pattern recognition for authentication of traceability data on foodstuffs, is the need for databases with analytical data on reference samples, covering the natural variation among the samples to be classified. Databases for marine oils need not be as extensive as for fish, since marine oils are generally produced from fish batches. Oils can also be analyzed directly without extensive sample preparation, and the greatest potential for official application may lie in the analysis of oils.

La déstabilisation de l'ordre antiferromagnétique de Néel au profit de nouvelles phases quantiques à température nulle à deux dimensions est envisageable grâce au phénomène de frustration magnétique. Le modèle théorique de spins Heisenberg S=1/2 répartis sur le réseau bidimensionnel frustré kagome, constitué de triangles joints uniquement par leurs sommets, est susceptible de stabiliser des phases quantiques originales de liquides de spin, qui ne présentent aucune brisure de symétrie à T = 0. Cette thèse a été consacrée à l'étude expérimentale de deux types de composés de spins S=1/2 (Cu2+) à géométrie kagome à l'aide de techniques spectroscopiques locales, la RMN et la μSR, ainsi que de mesures thermodynamiques (susceptibilité magnétique, chaleur spécifique). Dans Mg-herbertsmithite, la frustration est générée par une interaction d'échange premiers voisins antiferromagnétique J et est responsable d'un comportement liquide de spin jusqu'à des températures de l'ordre de J/10000. Par rapport au composé isostructural antérieur, Zn-herbertsmithite, nous avons montré qu'il possédait des propriétés physiques similaires tout en permettant une caractérisation fine du taux de défauts de substitutions Cu/Mg. Nos expériences réalisées à partir d'échantillons contrôlés permettent d'étudier finement l'origine des plateaux de relaxation observés en μSR à basse température en lien avec l'existence des défauts de spins interplans. La kapellasite et l'haydéite possèdent des interactions ferromagnétiques (J1) et antiferromagnétiques (Jd), offrant la possibilité d'explorer le diagramme de phases générées par la compétition de ces interactions sur le réseau kagome. Pour la kapellasite, nos mesures de μSR démontrent le caractère liquide de spin jusqu'à T ≈ J1/1000. La dépendance en température de la susceptibilité magnétique sondée par RMN du 35Cl ainsi que de la chaleur spécifique permettent d'évaluer le rapport Jd/J1 = 0.85, qui localise classiquement son fondamental au sein d'une phase originale de spins non coplanaires à 12 sous-réseaux appelée cuboc2. Les interactions présentes dans l'haydéite localisent son fondamental au sein de la phase ferromagnétique, en bon accord avec nos mesures qui indiquent une transition partielle à caractère ferromagnétique à T = 4 K. Cette étude confirme la pertinence du réseau kagome frustré pour la stabilisation de phases quantiques originales et démontre l'existence d'une nouvelle phase liquide de spin sur ce réseau, distincte de celle attendue pour des spins couplés antiferromagnétiquement.

The growing technique of near infrared spectroscopy is used to build fundamental knowledge of how the method might be applied for moisture determination in pulp and paper media. Filter paper was used to investigate the angle and distance dependence, otherwise obscured by the uneven surface of pulp or dry fibers. For contact measurements both filter paper and CTMP-fibers were used. The angle and distance evaluation is based on visual spectral interpretation to later incorporate preprocessing methods of the data followed by a multivariate analysis using PLS. With the knowledge built before the experimental startup, the acquired spectra are known to have many co-linearities and hence, for the purpose of determining the moisture content abundant information which might enable the reduction of data points for evaluation. The distance and angle experiments gave results indicating that the intensity of the reflected light change the information gathered and must be accounted for in calibration. Turning the measuring probe 30 degrees or more at 5 mm distance reduced the effect of high intensity specular reflection. Temperature displayed differences in the spectra, cold samples giving less response than warm samples although no clear correlation was found. Multivariate analysis of the contact measurement samples gave most accurate result after multiple signal correction preprocessing. Distance measurements on fiber rendered models with high RMSEC values and gave no reasonable base for prediction.

The advent of hyperpolarized magnetic resonance (MR) has provided new potential for real-time visualization of in vivo metabolic processes. The aim of the work in this thesis was to use hyperpolarized substrates to study rapid metabolic processes occurring in the healthy and diseased rat heart. Initial work, described in Chapter 2, optimized the hyperpolarization process to reproducibly generate tracers. Chapter 3 describes use of hyperpolarized 1-13C-pyruvate to investigate in vivo flux through the regulatory enzyme pyruvate dehydrogenase (PDH). Cardiac PDH activity was altered in several physiological and pathological states, namely fasting, type 1 diabetes, and high-fat feeding, and in vivo flux through PDH was measured using hyperpolarized MR. These measurements correlated with measurements of in vitro PDH activity obtained using a validated biochemical assay. The work in Chapter 4 investigated the physiological interaction between hyperpolarized tracer and cardiac tissue. The effect of hyperpolarized 1-13C-pyruvate concentration on its in vivo metabolism was analyzed using modified Michaelis-Menten kinetics. It was found that hyperpolarized MR could non-invasively follow mechanisms of metabolic regulation, in addition to reporting enzyme activity. In Chapter 5, hyperpolarized MR was incorporated into the isolated perfused rat heart. 1-13C-pyruvate in normal and ischaemic hearts revealed significant differences in lactate metabolism, and provided the foundation for a novel intracellular pH probe. Infusion of 2-13C-pyruvate in the isolated rat heart enabled the first real-time visualization of Krebs cycle intermediates. In summary, the work in this thesis has highlighted the potential of hyperpolarized MR to reveal novel information on heart disease.

L’emballage joue un rôle important dans l’industrie alimentaire afin de maintenir la qualité des produits le plus longtemps possible. Les films de polymère sont largement utilisés dans l’emballage alimentaire et sont attrayants pour leurs propriétés exceptionnelles. Puisque les polymères à base de pétrole apportent des préoccupations environnementales, les polymères biodégradables tel que le PLA sont étudiés plus intensivement depuis quelques années considérant leurs propriétés écologiques. L’application de films renforcés permet, comparativement aux films simples, d’atteindre des fonctions spécifiques et d’améliorer leurs propriétés tel que l’étanchéité aux gaz. Toutefois, puisque la structure de ces films est plus complexe, le contrôle de qualité de ces derniers est plus difficile. Dans l’industrie, des méthodes hors-ligne sont très souvent utilisées pour effectuer le contrôle de qualité des films produits. Le contrôle est nécessaire puisque la variabilité de la matière première ainsi que le changement des conditions opératoires amènent des modifications qui changent considérablement les propriétés du film. Par conséquent, une inspection en temps réel ainsi qu’un contrôle des films de polymère est nécessaire sur la ligne de production afin d’obtenir un contrôle de qualité s’approchant de l’analyse en temps réel. Un système d’imagerie proche infrarouge (NIR) rapide et non-destructif est proposé pour caractériser les films biodégradables d’acide polylactique contenant du talc produits par extrusion-soufflage et utilisés pour l’emballage. Le but ultime est d’utiliser le système pour faire un contrôle de qualité sur la ligne d’extrusion ainsi qu’après le post-traitement termique, soit le recuit. Un ensemble d’échantillon de film de PLA contenant différentes concentrations en talc ont été fabriqués. Ces derniers ont ensuite été soumis à différentes conditions de recuit. Des images NIR ont été collectées avant la caractérisation des propriétés physiques et mécaniques ainsi que l’étanchéité aux gaz. Des techniques d’imagerie multivariées ont été appliquées aux images hyperspectrales. Celles-ci ont montré que la quantité de talc peut être déterminée et que l’information du spectre NIR permet de prédire les propriétés du film. Dans tous les cas, la méthode proposée permet de déterminer les variations dans les propriétés du film avec une bonne précision.
Food packaging plays a great role in the food industry to maintain food products quality as long as possible. Polymer films are widely used in food packaging and also attract attention because of their outstanding advantages. Since petroleum-based polymers are known to cause environmental concerns, biodegradable polymers like PLA were studied more intensively in recent years due to their environmentally friendly properties. The application of reinforced films exceeds simple ones in achieving specific functions and enhancing their properties such as barrier properties. Since the films structures are more complex, quality control is more challenging. In industry, off-line methods are vastly used for quality control of the produced films while variability in raw materials and processing conditions substantially change the film specifications. Consequently, real-time inspection and monitoring of polymer films is needed on the production line to achieve a real-time quality control of the films. A fast and non-invasive near-infrared (NIR) imaging system is proposed to characterize biodegradable polylactic acid (PLA) films containing talc, and produced by extrusion film-blowing for packaging applications. The ultimate goal is to use the system for quality control on the extrusion line, and after a post-processing via thermal treatment (annealing). A set of PLA-talc films with varying talc contents were produced and submitted to annealing under different conditions. NIR images of the films were collected after which the samples were characterized for their physical, mechanical, and gas barrier properties. Multivariate imaging techniques were then applied to the hyperspectral images. It is shown that various talc loadings can be distinguished, and the information contained in the NIR spectra allows predicting the film properties. In all cases, the proposed approach was able to track the variation in film properties with good accuracy

Herbaceous perennials represent a considerable portion of potential biomass feedstocks available for the growing bioenergy industry. Their chemical composition and biomass yields, which are important in determining ethanol potential on an area and mass basis, vary with plant variety and type, environment, and management practices. Therefore, a study was conducted to assess the variability of lignin and carbohydrate content, biomass yields, and theoretical ethanol yields on an area basis among different herbaceous perennial species combinations grown in Minot (2008) and Williston (2008, 2009, and 2010), North Dakota (ND). After wet chemistry compositional analysis was done, the carbohydrate contents were used to determine theoretical ethanol potential on a mass basis. Using the dry-matter yield, the theoretical ethanol yield on an area basis was also calculated for these biomass species. Total carbohydrate content for the biomass samples in Williston and Minot varied from 45 to 61% dry basis. Analysis of Variance (ANOVA) at a= 0.05 showed that carbohydrate content varied between years and environments. Also an interaction plot shows that no biomass species had consistently higher or lower carbohydrate content in the different environments. Switchgrass (Panicum vigatum L.) grown as single species or together with other perennial grasses had higher dry-matter yield and theoretical ethanol yield potential in Williston irrigated plots while mixtures containing intermediate or tall wheatgrass species (Thinopyrum spp.) produced better yields in Minot non-irrigated plots. Variability in theoretical ethanol yield on a mass basis (3.7% coefficient of variation (CV) in Williston and 9.7% CV in Minot) was much less than the variability in dry-matter yields (27.5% CV in Williston and 14.8% CV Minot). Therefore, biomass production is much more important than composition in choosing species to grow for ethanol production. Recently, many studies have focused on developing faster methods to determine biomass composition using near infrared (NIR) spectroscopy. Other NIR models have been developed on single biomass feedstocks but a broad-based model for mixed herbaceous perennials is yet to be developed. Therefore, NIR calibration models for lignin, glucan, and xylan were developed with 65 mixed herbaceous perennial species using a DA 7200 NIR spectrometer (950 - 1,650 nm) and GRAMS statistical software. The models for lignin and xylan had R(2) values of 0.844 and 0.872, respectively, upon validation and are classified as good for quality assurance purposes while glucan model had an R(2) of 0.81 which is considered sufficient for screening. The R(2) and the root mean square error of prediction (RMSEP) results showed that it is possible to develop calibration models to predict chemical composition for mixed perennial biomass when compared with results for models developed for single feedstock by Wolfrum and Sluiter (2009) and Liu et al. (2010). Studying the variability in predicting constituents using NIR spectroscopy over time (hours and days), it was observed that the average CV was between 1.4 to 1.6%. The average CV due to repacking (presentation) alone was 1.3%. The CVs for NIR predictions ranged between 1.4 to 5.7% while for wet chemistry ranged between 3.8 to 13.5%; hence, NIR predictions were more precise than wet chemistry analysis.

La structure sous-jacente dans la zone A~100, N~60 a été étudié intensivement et extensivement, principalement par décroissance β et spectroscopie γ suite à des réactions de fission. Autour de N~60, en ajoutant juste quelques neutrons, protons un changement de forme rapide des états fondamentaux se produit, allant de sphérique à bien déformé. La coexistence de forme observée dans les noyaux de Sr et Zr est supposée avoir lieu dans toute la région. Les mécanismes impliqués dans l'apparition de la déformation n'étaient pas clairement identifiés. L'interaction entre les orbitales de Nilsson montantes et descendante est évoqué comme l'une des principales raisons du changement de forme. Cependant, une identification claire des orbitales proton et neutron en jeu était nécessaire. A cet effet, l'étude des isotopes ⁹³′⁹⁵′⁹⁷′⁹⁹Rb riches en neutrons a été réalisé excitation Coulombienne au CERN (ISOLDE) en utilisant le post-accélérateur REX-ISOLDE et le dispositif Miniball. Les structures excitées encore inconnues des isotopes ⁹⁷′⁹⁹Rb ont été peuplées et observées. Les coïncidences de transitions γ des états de basse énergie ont été observées et leur corrélations ont permis la construction de schémas de niveaux. Les probabilités de transitions associées ont été extraites grâce code GOSIA. Les éléments de matrice de l'opérateur électromagnétique observées constituent de nouveaux apports afin d'effectuer de nouveaux calculs théoriques permettant de statuer sur les orbitales impliquées. La sensibilité des expériences de ce type peut être accrue en utilisant des faisceaux radioactifs d'ions dont le spin nucléaire est polarisé. La technique de polarisation des feuilles orientées (TFT) fut étudiée dans ce but au CERN. Un nouveau polariseur TFT et un dispositif β-NMR ont être créés et installés après REX-ISOLDE. La connaissance du processus de polarisation associé à la technique reste incomplète à ce jour et de plus amples études sont nécessaires. Des tests préliminaires prometteurs ont été effectués sur le noyau de ⁸Li afin de déterminer le potentiel du dispositif actuel.

Theories of how galaxies form and evolve depend greatly on constraints provided by observations. However, when those observations come from different datasets, systematic offsets may occur. This causes difficulties measuring variations in parameters between filters. In this thesis I present the variation in total luminosity density with wavelength in the nearby Universe (z<0.1), produced from a consistent reanalysis of NIR and optical observations, taken from the MGC, UKIDSS and SDSS surveys. I derive luminosity distributions, best-fitting Schechter function parameterisations and total luminosity densities in ugrizYJHK, and compare the variation in luminosity density with cosmic star formation history (CSFH) and initial mass function (IMF) models. I examine the r band luminosity distribution produced using different aperture definitions, the joint luminosity- surface brightness (bivariate brightness) distribution in ugrizYJHK, comparing them to previously derived distributions, and how the total luminosity density varies with wavelength when surface brightness incompleteness is accounted for. I find the following results. (1) The total luminosity density calculated using a non-Sersic (e.g. Kron or Petrosian) aperture is underestimated by at least 15%, (2) Changing the detection threshold has a minor effect on the best-fitting Schecter parameters, but the choice of Kron or Petrosian apertures causes an offset between datasets, regardless of the filter used to define the source list, (3) The decision to use circular or elliptical apertures causes an offset in M* of 0.20 mag, and best-fitting Schechter parameters from total magnitude photometric systems have a flatter faint-end slope than Kron or Petrosian photometry, (4) There is no surface brightness distribution evolution with luminosity for luminous galaxies, but at fainter magnitudes the distribution broadens and the peak surface brightness dims. A Choloniewski function that is modified to account for this surface brightness evolution fits the bivariate-brightness distribution better than an unmodified Choloniewski function, (5) The energy density per unit interval, vf(v) derived using MGC and GAMA samples agrees within 90% confidence intervals, but does not agree with predictions using standard CSFH and IMF models. Possible improvements to the data and alterations to the theory are suggested.

Ce travail a pour objectif de contribuer a l'identification des resonances obtenues in vivo par spectroscopie rmn proton 1d et 2d, globale et localisee. Les techniques d'edition spectrale et de caracterisation developpees ont ete appliquees en particulier a l'etude des tumeurs intracerebrales du cerveau de rat in vivo. Un travail preliminaire a consiste a elaborer une methode rapide de correction des imperfections introduites par un modulateur de phase et d'amplitude, qui permet d'obtenir des impulsions de bonne qualite. Plusieurs axes de recherche ont ensuite ete suivis afin d'apporter des informations complementaires contribuant a l'identification moleculaire. Un de ces axes a concerne les techniques de rmn a deux dimensions, en examinant plus particulierement les fonctions d'apodisation utilisees pour le traitement des spectres 2d obtenus in vivo, afin d'optimiser le rapport signal sur bruit. La spectroscopie 2d de correlation combinee a l'imagerie spectroscopique a permis d'acceder a la repartition spatiale des acides amines et des carbohydrates presents dans une plante in vivo. D'autre part, les techniques d'observation localisees (imagerie et imagerie spectroscopique) des coherences a multiples quanta ont ete proposees afin d'observer simultanement les differents ordres de coherence des systemes de spins couples et non-couples. Enfin, dans un autre contexte, la mesure localisee du coefficient de diffusion a conduit a l'edition et a la caracterisation des metabolites presents dans les tumeurs intracerebrales in vivo en fonction de leur degre de mobilite translationnelle

Nous avons developpe une nouvelle methode d'exploration du metabolisme cerebral basee sur l'analyse du profil metabolique du lcr determine par spectroscopie de resonance magnetique (srm) du proton a haute resolution in vitro. Pour cela, nous avons mis au point un protocole d'analyse combinee du lcr et du serum par srm a haute resolution in vitro et hplc et developpe des programmes d'analyse automatique des spectres complexes de rmn a haute resolution. Les resultats sont ensuite traites par une analyse statistique en composantes principales. Nous montrons qu'il est ainsi possible de separer des groupes de pathologies neurologiques variees et qu'il existe une correlation entre l'eloignement de la representation graphique d'un individu par rapport a celle de la population controle et la severite de l'atteinte neurologique clinique. Nous avons effectue la premiere evaluation du metabolisme cerebral par srm du lcr in vitro au cours de pathologies degeneratives du systeme nerveux central (snc) chez l'homme: la maladie de huntington et les demences corticales degeneratives. Nous avons egalement realise la premiere exploration integree du metabolisme cerebral par srm a haute resolution du lcr et du serum in vitro combinee a la srm localisee cerebrale in vivo. Cette methode a ete appliquee a l'etude de la maladie de marchiafava-bignami et des complications neurologiques des deficits en cobalamine. Par sa plus grande sensibilite et l'identification plus aisee de nouveaux metabolites la srm du lcr et du serum in vitro autorise une meilleure comprehension des anomalies metaboliques detectees par la srm localisee cerebrale in vivo. Nos resultats demontrent l'interet, et la possibilite actuelle du developpement clinique de cette analyse integree du metabolisme cerebral par srm combinee in vitro et in vivo pour le diagnostic, le suivi therapeutique ou l'evaluation pharmacologique de nouvelles molecules dans diverses pathologies du snc chez l'homme

Ce travail porte sur la synthèse, la mise en forme et la caractérisation desolutions solides de type tysonite RE1-xMxF3-x (RE = La, Sm, Ce et M = Ba, Ca, Sr). Dans unpremier temps, une démarche d’étude rigoureuse est mise en place pour la solution solide ditede référence, La1-xBaxF3-x. Les synthèses menées à l’état solide aboutissent à une maîtrise dela composition chimique et à l'établissement de lois de variations des paramètres structuraux.Une meilleure compréhension de l’influence de la structure sur la mobilité des ions F- estégalement acquise. L’influence du frittage dans l’obtention de bonnes valeurs de conductivitéionique est également à souligner. Dans un second temps, les effets de la nanostructurationpar mécanobroyage sur les propriétés de conductivité sont évalués. L’utilisation de laméthodologie des plans d’expériences mène à la mise au point des réglages optimums debroyage. Il apparaît alors que la synthèse des électrolytes peut être accélérée et mise àl’échelle tout en gardant des valeurs optimales de conductivité. Enfin, la démarche déterminéeest appliquée à d'autres solutions solides de type tysonite et à la recherche du conducteurionique le plus performant. Si les composés issus de la substitution Ce/Sr ou encore Sm/Casemblent les plus prometteurs, la plus grande stabilité chimique de la solution solide La1-xSrxF3-x est le compromis idéal pour l'utiliser comme électrolyte solide lors des mesuresélectrochimiques des batteries
This work deals with the synthesis, shaping and characterization of RE1-xMxF3-x (RE = La, Sm, Ce et M = Ba, Ca, Sr) tysonite-type solid solutions. In a first part, onemeticulous approach has been set up for La1-xBaxF3-x solid solution, chosen as a reference.The solid-state synthesis of these materials led to a better knowledge of their chemicalcomposition (Vegard’s laws) and of the structure-ionic mobility correlations. The impact ofthe sintering process on the ionic conductivity is also highlighted. In a second part, the effectsof the nanostructuration conducted by ball-milling of the microcrystalline samples areevaluated. The use of the Design of Experiments methodology led to identify the optimummilling conditions. It appears that the synthesis of electrolytes can be sped- and scaled-up,while keeping high ionic conductivity properties. At last, this approach is applied on othertysonite-type solid solutions, to look for the best electrolyte. The Ce/Sr and Sm/Casubstitutions generate very promising ionic conductors but not really (electro)chemicallystable compounds. A compromise has been found with the choice of the La1-xSrxF3-x solidsolution as the FIB electrolyte for the electrochemical performances tests, regarding its higherchemical stability

The work reported in this thesis contains extensive studies carried out to investigate the weak molecular interactions in small organic molecules by utility of multinuclear and multidimensional NMR spectroscopic techniques and multitude of Density Functional based theoretical computations. The chemical shift of NH protons in benzoyl urea and the solvent dilution experiments infer the existence of dimers. The DOSY NMR spectroscopy conforms the existence of dimer formation in benzoyl urea derivatives. The formation of dimers is attributed to intermolecular HB and supported by DMSO titration studies. Along with the intermolecular HB, we have also observed the formation of intramolecular HB in all the dimers. The different 1D and 2D NMR experiments revealed the existence of both inter- and intra- molecular HB in all the synthesized derivatives of benzoyl urea. The existence of benzoyl ureas as dimer has been further confirmed by the DFT based computational analysis. The computational studies have been carried out by using QTAIM, NBO, NCI, EDD plots and MESP plots. In QTAIM analysis, we have discussed about Koch and Popelier criteria required for HB. Most of the criteria are satisfied by both inter- and intra- molecular HBs. The NCI analysis also revealed existence of inter- and intramolecular HB in all the synthesized derivatives of benzoyl urea. The NBO studies provide the information about the strength of HBs in terms of orbital stabilization energy. The EDD plots showed the formation of inter- and intra- molecular interactions in terms of electron density shifts involved in bonding. MESP plots drawn revealed the electrostatic nature of HB in all the analysed dimers.

碩士
國立成功大學
化學系
88
We have measured the values of nitrogen-14 quadrupole coupling constants (QCC), employing NMR relaxation techniques, for a designed series of para-substituted benzonitrile derivatives. It is found that for substituents bearing lone-paired electrons result in lower magnitudes of QCCs(14N) than the parent compound, benzonitrile. This observation provides solid evidence in support of the existence of “negative hyperconjugation” (NHC). For the methyl-substituted benzonitrile, the same observation is also present and can be rationalized by the well-known hyperconjugation effect. Although both effects may be viewed as the no-bond double-bond resonance based on valence bond theory, only NHC can be well demonstrated by “orbital interaction,” reported by Schleyer, within the molecular orbital theory. In cyanohalogens, the trend of QCCs(14N) cannot be accounted for in terms of the inductive effect reported for BrCN. On the other hand, it can be well understood on the NHC basis. Moreover, this trend of QCCs indicates that the -effect is shielded by the carbon atom and thus only pi-effect is the dominant factor affecting variations of QCC(14N) values. This is in accord with the “shielding effect” proposed much earlier in literature. More significantly, the NHC argument can be applied to account for the chain-conjugated structure proposed for sydnone compounds in the manner that the lone-paired electrons on the exocyclic oxygen, based on the Kartritzky hybrid structure, proceeds NHC into the pi-system of the N(2)-N(3)-C(4)-C(5) framework. Combined with the “ionic covalent bond” utilized for interpretation of the substantial stabilization of fluoro-substituted methanes, we conclude that the Kartritzky structure may also explain the IR and X-ray data observed for the highly polar C(5)-O(6) bond. It is then reasonable to exclude the aromaticity of the sydnone ring, whereas the semi-aromatic resonance structure is still favored from the viewpoint of NHC.

Thesis (M.Eng.)--University of Louisville, 2006.
Title and description from thesis home page (viewed Jan. 30, 2007). Department of Computer Engineering and Computer Science. "December 2006." Includes bibliographical references (p. 32-33).

This thesis presents results of novel methodologies applied to oriented systems. Both pure liquid crystalline materials as well as molecules oriented in liquid crystalline matrices have been studied. In particular this thesis presents investigations related to various aspects of NMR in liquid crystalline media, such as, assignment of resonances and the study of director dynamics of spinning liquid crystals in different phases and with different symmetry. Simplified methods for structure determination of solutes dissolved in liquid crystal solvents have been proposed. Diffusion ordered spectroscopy has been used to study a mixture of liquid crystals of opposite diamagnetic susceptibility at its coexistent phase. The methods presented represent novel techniques to characterize the liquid crystalline phase. NMR spectroscopy which has become a method of choice for understanding ordering mechanisms of mesogens requires a robust method for obtaining assignments of the NMR spectra of various nuclei that are found in the mesogens [1, 2]. It turns out that the spectra in the isotropic phase and in the nematic phase of a liquid crystal molecule are very different due to the presence of chemical shift anisotropy in the mesophase spectrum. There are a host of methodologies available for assigning spectra in the isotropic phase [3]. These methods however fail, when applied to the spectrum of the molecules in the mesophase due to the dominating role of strong anisotropic interactions, such as homonuclear couplings among protons. Problems arising while assigning spectral lines of liquid crystals in their nematic phase have been dealt with in chapter 2. To circumvent these problems, a property of the liquid crystal molecules under off-magic angle sample spinning can be utilized. It has been shown by Courtieu et al. [4] that the director/symmetry axis of a Δχ + ve liquid crystal aligns along the spinning axis for θ between 0 ° and θm, where θ is the angle between the spinning axis and the magnetic field and θm = 54.7° is the magic angle. It may be noted that the spectrum of θ = 0° spinning angle corresponds to the normal static spectrum, while the spectrum of θ = θm corresponds to the isotropic spectrum. In an earlier study, Teearr et al. [5] had recorded the 13C liquid crystal spectra as a function of very closely spaced θ values from 90° all the way up to 0°. From these plots of chemical shift versus the angle of spinning, it is possible to follow the trajectory of each 13C line from its position from θ = θm to θ = 0° and then match the spectrum in the isotropic phase (equivalently the magic angle sample spinning spectrum of the nematic phase) to the spectrum of the static sample in the nematic phase. However this method requires recording spectra at closely spaced angle intervals, so that one can unambiguously follow the trajectory of each of the lines without missing out any crossover of trajectories. However, this operation is time consuming. In this thesis we propose an alternate method, where we utilize the fact that the above trajectory has a very distinct relationship to the isotropic and anisotropic chemical shift and the problem of assignment does not require a continuous variation of angles, but just a few selected experiments should enable the assignment of the spectrum in the anisotropic phase. Thus the method of assignment has been made simpler and faster. It is shown that in addition to the assigned isotropic spectrum, only one other Off-magic angle spinning spectrum whose spinning angle θ is accurately known is necessary to obtain the complete assignment of the static spectrum. This procedure is non-trivial due to possibilities of errors in assignments arising out of inaccuracies in the knowledge of chemical shifts and the spinning angle. A computational procedure is proposed to take into account deviations arising out of non-ideal experimental conditions. A discussion regarding the details of the procedure and also situations where there can be ambiguities and how they can be resolved has been elaborated. The developed method has been demonstrated on a well known thermotropic liquid crystalline system, N-(4-ethoxybenzylidene)-4-n-butlyaniline [EBBA]. Since assignment of resonances in the nematic phase is a primary requirement for any further analysis regarding the ordering and deeper understanding of the role of various substituents in the mesogens we believe our novel prescription will be of immense use and utility. The third chapter presents the study of director dynamics in a lyotropic liquid crystal composed of Potassium laurate, 1-Decanol and D2O [6] under variable angle sample spinning using 2H NMR spectrum of D2O. A very interesting interplay of the magnetic orienting torque due to interaction of the liquid crystal director with the magnetic field and viscous torque arising from the viscosity of the sample on the director comes to fore. The relative magnitude of these torques has a direct bearing on the spectral pattern and line shapes observed, providing valuable insights into magnetohydrodynamics of the spinning liquid crystals. This study leads to even more interesting behavior for liquid crystals which deviate from uniaxial symmetry. This competition between magnetic and viscous torques has been quantitatively visualized by simulation of the 2H spectrum. It has been possible to visualize the observed spread in the director distribution arising out of viscous torque in terms of the energetics of the system under fast spinning. If the magnetic torque dominates over the viscous torque, then the equilibrium corresponds to the director orientation of δ = 0° where the energy is at its minimum. However the viscous and magnetic torques can become comparable as it may happen if the spinning angle is close to the magic angle or when the Δχ of the system is small. In those circumstances additional energy from the viscous torque causes the distribution of the director orientation to spread further away from δ = 0° for a positive Δχ liquid crystal. The trigonometric factor [P2(cosθ)∗P2(cosδ)] being proportional to the total energy of the system has been plotted against the spinning angle. The spectrum of the biaxial phase [7] as a function of the spinning angle shows more interesting director distribution. Here the patterns of the director distribution are observed on either side of the magic angle due to the presence of more than one director. The patterns observed also have information about the symmetry of the phase. This work provides insights into magnetohydrodynamics of spinning liquid crystals and can also be of relevance to samples of biological interest such as bicelles with protein oriented in them [8]. The fourth chapter deals with a novel characterization method relevant for the biaxial phase [9]. As an off shoot of the previous chapter, it effectively overcomes the disadvantages of the previous experimental methods which require simulation and line shape fitting to extract useful parameters. The chapter also presents the measurement of geometrical parameters of oriented solutes in phases exhibiting biaxial symmetry. The measured parameters show the effect of the onset of biaxiality as significant deviation in the value of the measured parameter. The utility of liquid crystalline media as solvents in high resolution NMR spectroscopy has been very rewarding since the pioneering work of Saupe and Englert [6]. The intramolecular interactions within solutes are only partially averaged. As a result one obtains a liquid like spectrum while at the same time very useful anisotropic interactions such as dipolar couplings, chemical shift anisotropies, quadrupolar couplings and anisotropic part spin-spin J couplings are extracted [10]. NMR spectra of molecules dissolved in thermotropic liquid crystals have long been used to obtain structural and orientational information. As the same time the complexity of the spectrum increases with the increase in the number of spins and the reduction in symmetry of the molecule, which can make the spectral analysis forbidding. Generally proton spectra have been used to obtain the geometry of the proton skeleton of the molecule and the information that includes dilute X nuclei such as 13C and 15N are available only from satellites which are buried in the intense proton spectrum. Different inequivalent dilute spins coupled to protons form different coupled spin systems in their natural abundance and appear as satellites in the proton spectra. Identification of transitions belonging to each of the spin system is essential to determine heteronuclear dipolar couplings, which is a formidable task. The fifth chapter deals with development of the techniques to obtain the complete structure of the dissolved molecules including nuclei other than protons in their natural abundance. The use of inverse experiments has been elaborated to overcome the problems of sensitivity and complexity for solute molecules having larger number of spins. In the present study using HSQC and HMQC experiments, we have selectively detected spectra of each inequivalent rare spin coupled to protons in pyrazine, pyrimidine and pyridazine dissolved in thermotropic Phase 4 and Phase 5 liquid crystal solvents. This way we could obtain enhancement in the intensity of satellites signals without the interference from the signals connected to the major isotopomers. Besides, we could resolve a complex spectrum into its sub-spectra corresponding to individual 13C and 15N isotopomers. This separation of the spectra corresponding to individual sub-spin systems makes analysis easy and helps analyze larger systems with higher number of spins and lower symmetry. Besides 1H-1H dipolar couplings, 13C-1H and 15N-1H dipolar couplings have been determined in natural abundance, thereby giving the complete dipolar coupling network between all the spins in the molecule. In this treatment pyrazine, pyrimidine and pyridazine have been used as examples of methodology developed. It is expected that the method will be of wider use for several other similar systems. Chapter six describes the diffusion ordered spectroscopic investigation [11] of a phase arising out of mixing together two liquid crystals having opposite signs of diamagnetic susceptibility anisotropy [12]. Towards this end we have used CH3CN as a probe molecule. The spectrum of CH3CN has with it the information about the parallel or perpendicular orientation of the phase. Such a mixture of liquid crystals have shown interesting behavior at the critical temperature where the two phases seem to coexist. It has been an interesting question to understand what exactly happens for the molecular orientation when the macroscopic anisotropy Δχ vanishes. Earlier Jokisaari et al. [13] have varied the temperature very finely taking due precautions to maintain homogeneity and stability of temperature to the tune of ±0.05K across the sample volume. Their observation of a powder pattern exactly in the critical temperature was interpreted as arising out of a distribution of directors equally oriented in all directions. In our experiments we have measured the diffusion coefficient of the probe molecule i.e. acetonitrile as we change the temperature of the system through the critical temperature. At the critical temperature we have a situation of being able to measure the parallel and perpendicular orientational diffusion coefficients simultaneously. The measurements show that the parallel component of the diffusion coefficient has reduced and the perpendicular component has increased in comparison to the trend in the immediate neighboring temperatures, thereby indicating that at the exact critical condition the liquid crystal mixture consists of an isotropic distribution of molecules. As a check to rule out any exchange of molecules in different domains of parallel and perpendicular orientations an EXSY experiment was conducted with a mixing time which was same as that of the diffusion delay in the DOSY experiment. The EXSY spectrum showed no exchange cross peaks between the two orientations, this confirms that the anisotropy of the diffusion vanishes at the critical temperature. Nematic liquid crystals exhibit a rich variety of phases and properties. NMR is a very powerful tool to study the various phases at the microscopic and molecular level. It has also turned out that some of these properties can be usefully utilized for investigation of both small and large molecules by NMR. Thus this thesis has attempted to expand several of the techniques already available for various applications and extend the utility of NMR for the study of partially ordered systems.

This thesis presents results of novel methodologies applied to oriented systems. Both pure liquid crystalline materials as well as molecules oriented in liquid crystalline matrices have been studied. In particular this thesis presents investigations related to various aspects of NMR in liquid crystalline media, such as, assignment of resonances and the study of director dynamics of spinning liquid crystals in different phases and with different symmetry. Simplified methods for structure determination of solutes dissolved in liquid crystal solvents have been proposed. Diffusion ordered spectroscopy has been used to study a mixture of liquid crystals of opposite diamagnetic susceptibility at its coexistent phase. The methods presented represent novel techniques to characterize the liquid crystalline phase. NMR spectroscopy which has become a method of choice for understanding ordering mechanisms of mesogens requires a robust method for obtaining assignments of the NMR spectra of various nuclei that are found in the mesogens [1, 2]. It turns out that the spectra in the isotropic phase and in the nematic phase of a liquid crystal molecule are very different due to the presence of chemical shift anisotropy in the mesophase spectrum. There are a host of methodologies available for assigning spectra in the isotropic phase [3]. These methods however fail, when applied to the spectrum of the molecules in the mesophase due to the dominating role of strong anisotropic interactions, such as homonuclear couplings among protons. Problems arising while assigning spectral lines of liquid crystals in their nematic phase have been dealt with in chapter 2. To circumvent these problems, a property of the liquid crystal molecules under off-magic angle sample spinning can be utilized. It has been shown by Courtieu et al. [4] that the director/symmetry axis of a Δχ + ve liquid crystal aligns along the spinning axis for θ between 0 ° and θm, where θ is the angle between the spinning axis and the magnetic field and θm = 54.7° is the magic angle. It may be noted that the spectrum of θ = 0° spinning angle corresponds to the normal static spectrum, while the spectrum of θ = θm corresponds to the isotropic spectrum. In an earlier study, Teearr et al. [5] had recorded the 13C liquid crystal spectra as a function of very closely spaced θ values from 90° all the way up to 0°. From these plots of chemical shift versus the angle of spinning, it is possible to follow the trajectory of each 13C line from its position from θ = θm to θ = 0° and then match the spectrum in the isotropic phase (equivalently the magic angle sample spinning spectrum of the nematic phase) to the spectrum of the static sample in the nematic phase. However this method requires recording spectra at closely spaced angle intervals, so that one can unambiguously follow the trajectory of each of the lines without missing out any crossover of trajectories. However, this operation is time consuming. In this thesis we propose an alternate method, where we utilize the fact that the above trajectory has a very distinct relationship to the isotropic and anisotropic chemical shift and the problem of assignment does not require a continuous variation of angles, but just a few selected experiments should enable the assignment of the spectrum in the anisotropic phase. Thus the method of assignment has been made simpler and faster. It is shown that in addition to the assigned isotropic spectrum, only one other Off-magic angle spinning spectrum whose spinning angle θ is accurately known is necessary to obtain the complete assignment of the static spectrum. This procedure is non-trivial due to possibilities of errors in assignments arising out of inaccuracies in the knowledge of chemical shifts and the spinning angle. A computational procedure is proposed to take into account deviations arising out of non-ideal experimental conditions. A discussion regarding the details of the procedure and also situations where there can be ambiguities and how they can be resolved has been elaborated. The developed method has been demonstrated on a well known thermotropic liquid crystalline system, N-(4-ethoxybenzylidene)-4-n-butlyaniline [EBBA]. Since assignment of resonances in the nematic phase is a primary requirement for any further analysis regarding the ordering and deeper understanding of the role of various substituents in the mesogens we believe our novel prescription will be of immense use and utility. The third chapter presents the study of director dynamics in a lyotropic liquid crystal composed of Potassium laurate, 1-Decanol and D2O [6] under variable angle sample spinning using 2H NMR spectrum of D2O. A very interesting interplay of the magnetic orienting torque due to interaction of the liquid crystal director with the magnetic field and viscous torque arising from the viscosity of the sample on the director comes to fore. The relative magnitude of these torques has a direct bearing on the spectral pattern and line shapes observed, providing valuable insights into magnetohydrodynamics of the spinning liquid crystals. This study leads to even more interesting behavior for liquid crystals which deviate from uniaxial symmetry. This competition between magnetic and viscous torques has been quantitatively visualized by simulation of the 2H spectrum. It has been possible to visualize the observed spread in the director distribution arising out of viscous torque in terms of the energetics of the system under fast spinning. If the magnetic torque dominates over the viscous torque, then the equilibrium corresponds to the director orientation of δ = 0° where the energy is at its minimum. However the viscous and magnetic torques can become comparable as it may happen if the spinning angle is close to the magic angle or when the Δχ of the system is small. In those circumstances additional energy from the viscous torque causes the distribution of the director orientation to spread further away from δ = 0° for a positive Δχ liquid crystal. The trigonometric factor [P2(cosθ)∗P2(cosδ)] being proportional to the total energy of the system has been plotted against the spinning angle. The spectrum of the biaxial phase [7] as a function of the spinning angle shows more interesting director distribution. Here the patterns of the director distribution are observed on either side of the magic angle due to the presence of more than one director. The patterns observed also have information about the symmetry of the phase. This work provides insights into magnetohydrodynamics of spinning liquid crystals and can also be of relevance to samples of biological interest such as bicelles with protein oriented in them [8]. The fourth chapter deals with a novel characterization method relevant for the biaxial phase [9]. As an off shoot of the previous chapter, it effectively overcomes the disadvantages of the previous experimental methods which require simulation and line shape fitting to extract useful parameters. The chapter also presents the measurement of geometrical parameters of oriented solutes in phases exhibiting biaxial symmetry. The measured parameters show the effect of the onset of biaxiality as significant deviation in the value of the measured parameter. The utility of liquid crystalline media as solvents in high resolution NMR spectroscopy has been very rewarding since the pioneering work of Saupe and Englert [6]. The intramolecular interactions within solutes are only partially averaged. As a result one obtains a liquid like spectrum while at the same time very useful anisotropic interactions such as dipolar couplings, chemical shift anisotropies, quadrupolar couplings and anisotropic part spin-spin J couplings are extracted [10]. NMR spectra of molecules dissolved in thermotropic liquid crystals have long been used to obtain structural and orientational information. As the same time the complexity of the spectrum increases with the increase in the number of spins and the reduction in symmetry of the molecule, which can make the spectral analysis forbidding. Generally proton spectra have been used to obtain the geometry of the proton skeleton of the molecule and the information that includes dilute X nuclei such as 13C and 15N are available only from satellites which are buried in the intense proton spectrum. Different inequivalent dilute spins coupled to protons form different coupled spin systems in their natural abundance and appear as satellites in the proton spectra. Identification of transitions belonging to each of the spin system is essential to determine heteronuclear dipolar couplings, which is a formidable task. The fifth chapter deals with development of the techniques to obtain the complete structure of the dissolved molecules including nuclei other than protons in their natural abundance. The use of inverse experiments has been elaborated to overcome the problems of sensitivity and complexity for solute molecules having larger number of spins. In the present study using HSQC and HMQC experiments, we have selectively detected spectra of each inequivalent rare spin coupled to protons in pyrazine, pyrimidine and pyridazine dissolved in thermotropic Phase 4 and Phase 5 liquid crystal solvents. This way we could obtain enhancement in the intensity of satellites signals without the interference from the signals connected to the major isotopomers. Besides, we could resolve a complex spectrum into its sub-spectra corresponding to individual 13C and 15N isotopomers. This separation of the spectra corresponding to individual sub-spin systems makes analysis easy and helps analyze larger systems with higher number of spins and lower symmetry. Besides 1H-1H dipolar couplings, 13C-1H and 15N-1H dipolar couplings have been determined in natural abundance, thereby giving the complete dipolar coupling network between all the spins in the molecule. In this treatment pyrazine, pyrimidine and pyridazine have been used as examples of methodology developed. It is expected that the method will be of wider use for several other similar systems. Chapter six describes the diffusion ordered spectroscopic investigation [11] of a phase arising out of mixing together two liquid crystals having opposite signs of diamagnetic susceptibility anisotropy [12]. Towards this end we have used CH3CN as a probe molecule. The spectrum of CH3CN has with it the information about the parallel or perpendicular orientation of the phase. Such a mixture of liquid crystals have shown interesting behavior at the critical temperature where the two phases seem to coexist. It has been an interesting question to understand what exactly happens for the molecular orientation when the macroscopic anisotropy Δχ vanishes. Earlier Jokisaari et al. [13] have varied the temperature very finely taking due precautions to maintain homogeneity and stability of temperature to the tune of ±0.05K across the sample volume. Their observation of a powder pattern exactly in the critical temperature was interpreted as arising out of a distribution of directors equally oriented in all directions. In our experiments we have measured the diffusion coefficient of the probe molecule i.e. acetonitrile as we change the temperature of the system through the critical temperature. At the critical temperature we have a situation of being able to measure the parallel and perpendicular orientational diffusion coefficients simultaneously. The measurements show that the parallel component of the diffusion coefficient has reduced and the perpendicular component has increased in comparison to the trend in the immediate neighboring temperatures, thereby indicating that at the exact critical condition the liquid crystal mixture consists of an isotropic distribution of molecules. As a check to rule out any exchange of molecules in different domains of parallel and perpendicular orientations an EXSY experiment was conducted with a mixing time which was same as that of the diffusion delay in the DOSY experiment. The EXSY spectrum showed no exchange cross peaks between the two orientations, this confirms that the anisotropy of the diffusion vanishes at the critical temperature. Nematic liquid crystals exhibit a rich variety of phases and properties. NMR is a very powerful tool to study the various phases at the microscopic and molecular level. It has also turned out that some of these properties can be usefully utilized for investigation of both small and large molecules by NMR. Thus this thesis has attempted to expand several of the techniques already available for various applications and extend the utility of NMR for the study of partially ordered systems.