The thesis entitled “Exploring Diverse Facets of Small Molecules by NMR Spectroscopy” consists of six chapters. The main theme of the thesis is to exploit one and two dimensional NMR methodologies for understanding the diverse facets of small organic molecules, such as, weak intra- and inter- molecular interactions, chiral discrimination, quantification of enantiomeric excess and assignment of absolute configuration. Several new pulse sequences have also been designed to solve specific chemical problems, in addition to extensive utility of existing one and two dimensional NMR experiments. The results obtained on different problems, are discussed under six chapters in the thesis. The brief summary of each of these chapters is given below. Chapter 1 begins with the discussion on the importance of small molecules and their various facets, the analytical techniques available in the literature to study them. The role of NMR spectroscopy as powerful analytical technique to understand the diverse facets of organic molecules and their importance is set out in brief. A short introduction to the basic principles of NMR, the interaction parameters, the commonly employed one and two dimensional homo- and herero- nuclear NMR experiments are also given. The basic introduction to product operators essential for understanding the spin dynamics in the developed pulse sequences is given. The application of diffusion ordered spectroscopy (DOSY), the general problems encountered in the analysis of combinatorial mixtures and the matrix assisted method in circumventing such problems are discussed. Chapter 2 focuses on the chiral discrimination and the measurement of enatiomeric excess. The NMR approach to discriminate enantiomers using chiral auxiliaries such as, solvating agents, derivatizing agents, lanthanide shift reagents, the choice of such auxiliaries and the limitations are discussed in detail. The in-depth discussion on the new protocols developed using both the solvating and derivatizing agents for enantiomeric discrimination of chiral amines, hydroxy acids and diacids are discussed. The new three-component protocols that serve as chiral derivatizing agents for the discrimination of primary amines, diacids and hydroxy acids are discussed. Also the role of organic base such as DMAP in the chiral discrimination is explored for discrimination of acids using BINOL as a chiral solvating agent. Accordingly the discussion is classified into two sections. In the first section the protocol developed utilizing an enantiopure mandelic acid, a primary amine substrate and 2-formylphenylboronic acid that is ideally suited for testing the enantiopurity of chiral primary amines is discussed. The broad applicability of the protocols for testing enantiopurity has been demonstrated on number of chiral molecules using 1H and 19F NMR. The second section contains the results on the new concept developed for discrimination of hydroxy acids. The strategy involves the formation of three component protocol using chiral hydroxy acid, R-alphamethylbenzylamine and 2-formylphenylboronic acid for 1H-NMR discrimination of diacids. The section also includes the utility of ternary ion-pair complex for the discrimination of acids. The ternary ion-pair not only permitted the testing of enantiopurity of chiral acids, but is also found useful for the measurement of enantiomeric excess. Chapter 3 discusses the utilization of the developed three-component protocols for the assignment of absolute configurations of molecules of different functionality. The protocols for the assignments of absolute configuration of primary amines using 2-formylphenylboronic acid and mandelic acid yielded the substantial chemical shift differences between diastereomers. The consistent trend in the direction of change of chemical shifts of the discriminated proton(s) gave significant evidence for employing them as parameters for the assignment of spatial configuration of primary amines. Another protocol using 2-formylphenylboronic acid, hydroxy acids and enantiopure alphamethylbenzylamine permitted their configurational assignment. In the second section a novel solvating agent, obtained by the formation of an ion-pair complex among enantiopure BINOL, DMAP and chiral hydroxy acid for the assignment of the spatial configuration of hydroxy acids is discussed. Chapter 4 focuses on the development of novel NMR methodologies, and also the utility of existing two-dimensional experiments for addressing certain challenging problems. This chapter has been divided into three sections. In Section-I the utilization of well-known homonuclear 2D-J-resolved methodology for unravelling the overlapped NMR spectra of enantiomers, an application for chiral discrimination and the measurement of enantiomeric excess is discussed. The utilization of the chiral auxiliaries, such as, chiral derivatizing agents, chiral solvating agents and lanthanide shift reagents permits enantiodiscrimination and the measurement of excess of one form over the other. Nevertheless many a times one encounters severe problems due to small chemical shift difference, overlap of resonances, complex multiplicity pattern because of the presence of number of interacting spins, and enormous line broadening due to paramagnetic nature of the metal complex. This section is focused on combating such problems utilizing 2D-J-1JNH resolved spectroscopy where a 450 tilting of the spectrum in the F2 dimension, yielded the pure shift NMR spectrum. The method circumvents several problems involved in chiral discrimination and allows the accurate measurement of enantiomeric excess. In Section-II, the development of novel NMR experimental methodology cited in the literature as C-HetSERF and its application for the study of symmetric molecules, such as, double bonded cis- and trans- isomers, and extraction of magnitudes and signs of long range homo- and hetero- nuclear scalar couplings among chemically equivalent protons in polycylic aromatic hydrocarbons is discussed. The extensive utility of the new pulse sequence has been demonstrated on number of symmetric molecules, where the conventional one dimensional experiment fails to yield spectral parameters. In section III, yet another novel pulse sequence called RES-TOCSY developed for unravelling of the overlapped NMR spectrum of enantiomers and the measurement of enantiomeric contents, has been utilized for the accurate measurement of magnitudes and signs of 1H-19F couplings in fluorine containing molecules. The method has distinct advantages as the strengths of the couplings and their relative signs could be extracted on diverse situations, such as, couplings smaller than line widths, the spectrum where the coupling fine structures are absent. Chapter 5 covers the study of nature of intra- and inter- molecular hydrogen bond in amide and its derivative. The chapter is accordingly divided into two sections. In the first section the study of acid and amide hydrogen bonding is discussed and the hydrogen bonded interactions are probed by extensive utility of 1H, 13C and 15N-NMR. The temperature perturbation experiments, measurements of the variation in the couplings, monitoring of diffusion coefficients and the association constants, detection of through space correlation have given unambiguous evidence for the hydrogen bond formation. The results were also supported by DFT calculations. Similar interaction in the solid state has also been derived by obtaining the crystal structure of complex phenylacetic acid with benzamide. In the second section of the chapter the hydrogen bond interaction of organic fluorine in trifluoromethyl derivatives of benzanilides has been explored and the involvement of CF3 group in the hydrogen bonding has been detected. The evidence for the participation of CF3 group in hydrogen bond has been confirmed by number of experiments, such as, the detection of through space couplings, viz., 1hJFH, 1hJFN, and 2hJFF , where the spin polarization between the interacting spins is transmitted through hydrogen bond, the temperature and solvent dependent studies, variation in the 1JNH and two dimensional heteronuclear correlation experiments. In an interesting example of a molecule containing two CF3 groups situated on two phenyl rings of benzanilide, the simultaneous participation of fluorines of two CF3 groups in hydrogen bond has been detected. The confirmatory evidence for such an interaction, where hydrogen bond mediated couplings are not reflected in the NMR spectrum, has been derived by 19F−19F NOESY. Significant deviations in the strengths of 1JNH, in addition to variable temperature, and the solvent induced perturbation studies yielded additional evidence. The NMR results are corroborated by both DFT calculations and MD simulations, where the quantitative information on different ways of involvement of fluorine in two and three centered hydrogen bonds, their percentage of occurrences, and geometries have been obtained. The hydrogen bond interaction energies have also been calculated. The study revealed the rare observation and the first example of the C-F…H-N hydrogen bond in solution state in the molecules containing CF3 groups. Chapter 6 focuses on the mixture analysis using the diffusion ordered spectroscopy (DOSY). High Resolution-DOSY works when the NMR spectrum is well resolved and the diffusion coefficients of the combinatorial mixtures are substantially different from each other. DOSY technique fails when the mixture contains the molecules of nearly identical weights and similar hydrodynamic radii. Thus, the positional isomers, enantiomers consequent to their nearly identical rates of diffusion, are not differentiated. Some of these problems can be overcome by Matrix-Assisted Diffusion Order Spectroscopy (MAD-spectroscopy), where an external reagent acts as a matrix and aids in their diffusion edited separation, provided the molecules embedded in it possess differential binding abilities with the matrix. Such different binding properties of the matrix are the basis for resolution of many isomeric species. In the present study three different novel auxiliaries, micelles-reverse micelles, crown ether and cyclodextrin are introduced for the resolution of positional isomers, double bonded isomers, viz., fumaric acid and maleic acid and also enantiomers. Accordingly, the results of each of these studies are discussed in three different sections.

L’objectif principal de cette thèse est de caractériser l’atmosphère de Jupiters chaudes par spectroscopie de transmission à haute résolution, dans l’infrarouge proche, avec l’instrument SPIRou. L’historique de formation, d’évolution et de migration des planètes est empreint dans leur composition chimique, et de retrouver cette composition permet d’en élucider le mystère. La spectroscopie de transit et d’émission a prouvé être fortement efficace à cette tâche, autant pour la détermination de la composition que pour la détermination d’autres caractéristiques atmosphériques comme le profil de température et la dynamique, accessibles à haute résolution. Les Jupiter Chaudes, planètes géantes gazeuses qui orbitent très près de leur étoile, offrent des conditions d’observations très favorables pour ce type d’étude. Encore beaucoup d’éléments nous échappent quant aux processus physiques, chimiques et dynamiques qui gouvernent l’atmosphère de ces objets astronomiques. Des études détaillées de ceux-ci, telles que celles présentées dans cette thèse, sont nécessaires pour mieux comprendre ces mécanismes. Dans un premier temps, nous avons fait l’étude de deux transits de HD 189733 b, une des exoplanètes les plus étudiées. Cela nous a permis de valider nos méthodes d’analyse avec des données provenant du spectropolarimètre infrarouge à haute résolution SPIRou, installé au télescope Canada-France-Hawaii, représentant d’ailleurs la première caractérisation d’une atmosphère en spectroscopie de transit pour le SPIRou Legacy Survey. Nous avons confirmé la présence d’un signal d’eau à un niveau de signification de plus de 5σ, basé sur les résultats de test-t. Nous présentons la première analyse de spectroscopie de transit haute résolution faite par méthode de récupération, basée sur l’inférence bayésienne et appliquée à une grille de modèles SCARLET interpolée. Celle-ci a permis d’inférer une abondance de log_10[H2O] ≃ −4.4. Les résultats obtenus sont cohérents avec la littérature et indiquent que l’atmosphère de HD 189733 b est relativement claire (sans nuages) et possède un C/O super-solaire (correspondant à une formation au-delà de la ligne de glace de l’eau). Un fort décalage vers le bleu de l’absorption par l’eau a été observé, indiquant la présence de forts vents allant du côté jour vers le côté nuit ou un signal dominé par le côté soir du terminateur (limbe arrière), ou une combinaison des deux. Dans un second temps, nous présentons la première analyse à haute résolution dans l’infrarouge proche de trois transits de la très peu dense sous-Saturne WASP-127 b. Une étude récente à basse résolution a montré un spectre de transmission riche et une abondance super-solaire de CO2 dans son atmosphère. La contribution de CO et de CO2 n’a cependant pu être démêlée étant donné la couverture spectrale et la résolution limitée des données HST et de Spitzer utilisées, menant à des scénarios équiprobables de faible et fort C/O. La couverture de la bande de CO à 2.3 μm par SPIRou a permis de faire cette différenciation, et nos résultats ont exclu la présence de CO en abondance supérieure à log_10[CO] = −4.3, impliquant que le signal détecté à 4.5μm dans les données Spitzer provient majoritairement du CO2. De plus, un calcul de test-t sur les données SPIRou a confirmé la détection de H2O à un niveau ajusté de 4.9σ, mais également une détection potentielle de OH, à 2.4σ. Cette présence extrêmement inattendue de OH pourrait potentiellement être expliquée si la température du côté jour est assez grande, aidée par l’irradiation amplifiée de l’étoile qui quitte la séquence principale, ou encore par du mélange vertical. Nous présentons également la première méthode de récupération complète appliquée à la spectroscopie de transit à haute résolution, en utilisant la suite de modèles petitRADTRANS, et sur trois ensembles de données différents : les données SPIRou, les données HST et Spitzer de l’étude initiale, et les deux ensembles de données combinés. Une comparaison des différents résultats obtenus confirme que l’analyse conjointe permet d’avoir de meilleures contraintes sur les paramètres atmosphériques. Alors que l’étude initiale favorisait un fort C/O, nos résultats pointent vers un C/O très sous-solaire, produit par un C/H sous-solaire et un O/H plutôt stellaire. Les scénarios de formation qui supportent une telle composition sont ceux au-delà des lignes de glace de H2O et de CO2 (∼ 10ua), avec une accrétion supplémentaire de matériel riche en O via la migration et le croisement des lignes de glaces. L’accrétion du matériel est soit dominée par le gaz et tardive (après 5 à 7 millions d’années), ou encore, mixte (de gaz et de glaces) et plus précoce, avec un mélange cœur-enveloppe substantiel. Bien qu’il en reste beaucoup à faire, ces travaux de recherche ont démontré que la spectroscopie de transit à haute résolution dans l’infrarouge proche est utile pour explorer les conditions atmosphériques des Jupiter et sous-Saturne chaudes, et plus spécifiquement, avec l’instrument SPIRou. La combinaison de données à faible et à haute résolution s’avère un outil très puissant pour l’étude des atmosphères, et le sera encore plus avec les capacités révolutionnaires de JWST. / The main objective of this thesis is to characterize the atmosphere of hot Jupiters with high resolution transmission spectroscopy in the near-infrared with the SPIRou instrument. The formation, evolution and migration history of exoplanets is imprinted in their chemical composition, and finding this composition makes it possible to trace back this history. Transit and emission spectroscopy have proven to be highly effective for this task, in the determination of the composition, but also of other atmospheric characteristics such as the temperature profile and the dynamics, accessible at high resolution. Hot Jupiters -- gas giant planets orbiting very close to their star -- offer highly favourable observation conditions for this type of study. Many pieces of the puzzle are still missing regarding the physical, chemical and dynamical processes that govern the atmospheres of these astronomical objects, and detailed studies, such as the ones presented in this thesis, are necessary to better understand those mechanisms. First, we present the results of our analysis of two HD189733b transits, one of the most studied exoplanets to date. This study allowed to validate our analysis method with SPIRou data, a high-resolution near-infrared spectro-polarimeter installed at the Canada-France-Hawaii Telescope. It also represents the first characterization of an atmosphere with transit spectroscopy as part of the SPIRou Legacy Survey. Our results confirmed the H2O detection in the planet's atmosphere at a 5 sigma level, based on a $t$-test. We present the first analysis of a Bayesian retrieval framework applied to high-resolution transmission spectroscopy, using a grid of SCARLET models. We constrained the H2O abundance to log_10[H2O] ~ -4.4$. The results are consistent with the literature and agree on the atmosphere of HD189733b being relatively clear (without clouds) and having a super-solar C/O (corresponding to a formation beyond the H2O ice line). A strong blue shift of the water absorption signal was observed, indicative of strong day-to-night winds or a signal dominated by the terminator's evening side (trailing limb), or a combination of both. Second, we present the first high-resolution analysis in the near-infrared of three transits of the super low-density sub-Saturn WASP-127b. A recent low-resolution study showed a rich transmission spectrum and super-solar abundance of CO2 in its atmosphere. However, the contribution of CO and CO2 could not be disentangled given the limited spectral coverage and resolution of the HST and Spitzer data, leading to equiprobable low and high C/O scenarios. The coverage of the CO band at 2.3um by SPIRou made it possible to differentiate between the two cases, and our results excluded CO abundances greater than 10^(-4.3), implying that the signal at 4.5um seen in the Spitzer data mostly comes from CO2. Moreover, a t-test analysis on the SPIRou data confirmed the detection of H2O at an adjusted level of 4.9 sigma, but also a tentative detection of OH, at 2.4 sigma. The presence of OH, although extremely unexpected, could potentially be explained from a high enough dayside temperature, increased by the amplified irradiation of the star leaving the main sequence, or from vertical mixing. We also present the first full-retrieval framework applied to transmission spectroscopy at high resolution, using the petitRADTRANS model suite on three different datasets: on the SPIRou data, on the HST and Spitzer data from the original study, and on both datasets combined. A comparison of the different results obtained confirms that the joint analysis provides better constraints on the atmospheric parameters. While the initial study favored a high C/O, our results point toward a highly subsolar C/O, produced by a sub-stellar C/H and a roughly stellar O/H. Formation scenarios that support such a composition are those beyond the H2O and CO2 ice lines (~ 10ua), with further accretion of O-rich material via migration and ice lines crossing. The primordial/bulk accretion was either gas-dominated and late (after 5-7 Myr), or earlier and mixed (with gas and ice), with substantial core-enveloppe mixing. Although much remains to be done, this research work has demonstrated that high-resolution near-infrared transit spectroscopy is useful for exploring the atmospheric conditions of hot Jupiters and sub-Saturns, and more specifically, with the SPIRou instrument. The combination of low and high resolution data has shown to be a very powerful tool for such studies, and will be even more so with the revolutionary capabilities of JWST.