Two-dimensional Correlation Experiments Research Articles

A combined solid-state NMR and quantum chemical calculation study of hydrogen bonding in two forms of α-d-glucose

Hydrogen bonding plays an important role in the structure and function of a wide range of materials. Solid-state 1H nuclear magnetic resonance (NMR) spectroscopy provides a very sensitive tool to investigate the local structure of hydrogen atoms involved in hydrogen bonding. While there is extensive 1H solid-state NMR data on O–H - - O hydrogen bonding in solid carboxylic acids, there has been no systematic 1H solid-state NMR studies of hydroxyl groups in carbohydrates (and hydroxyl groups in general). With a view to studying the hydrogen bonding in more complex materials such as cellulose polymorphs, we carried out a detailed solid-state 1H NMR investigation of the model compounds α-d-glucose and α-d-glucose monohydrate. Through a combination of fast magic-angle spinning (MAS), combined rotation and multiple pulse spectroscopy (CRAMPS), and two-dimensional (2D) correlation experiments carried out at ultrahigh magnetic fields, it was possible to assign all of the aliphatic (CH), hydroxyl (OH), and water (H2O) 1H chemical shifts in both forms of α-d-glucose. Plane-wave DFT calculations were employed to improve the hydrogen atom positions for α-d-glucose monohydrate and to calculate 1H chemical shifts, providing additional support for the experimentally determined peak assignments. Finally, the relationship between the hydroxyl 1H chemical shifts and their hydrogen bonding geometry was investigated and compared to the well-established relationship for carboxylic acid protons.

Selective synthesis of A-ring Е-arylidene derivatives from β-sitosterol and their activity

A series of 24-ethylcholest-4-ene-3,6-dione 2E-arylidene-derivatives has been synthesized by a Claisen-Schmidt reaction from a natural phytosterol β-sitosterol with yields of 80–85%. The structure of the obtained compounds was confirmed by NMR spectroscopy, including two-dimensional correlation experiments. The synthesized compounds were evaluated for their in vitro cytotoxicity and α-glucosidase inhibitory activity. It was established that compound 3 with pyridin-3-ylmethylene moiety exhibited a selective cytotoxic effect against the U251 cancer cell line with 99.31% inhibition of cancer cell growth. Compounds with pyridin-4-ylmethylene 4 and furan-2-ylmethylene-5 fragments were the most active inhibitors of α-glucosidase with IC50 64.00 and 38.95 µM, being 3- and 5-times more active than acarbose. Binding mode to α-glucosidase and ADMET characteristics for the lead molecule 5 were proposed computationally. To sum up, an efficient approach to the derivatives with promising antidiabetic activity based on available natural product β-sitosterol is suggested.

Complete resonance assignment of a pharmaceutical drug at natural isotopic abundance from DNP-Enhanced solid-state NMR

Solid-state dynamic nuclear polarization enhanced magic angle spinning (DNP-MAS) NMR measurements coupled with density functional theory (DFT) calculations enable the full resonance assignment of a complex pharmaceutical drug molecule without the need for isotopic enrichment. DNP dramatically enhances the NMR signals, thereby making possible previously intractable two-dimensional correlation NMR spectra at natural abundance. Using inputs from DFT calculations, herein we describe a significant improvement to the structure elucidation process for complex organic molecules. Further, we demonstrate that a series of two-dimensional correlation experiments, including 15N–13C TEDOR, 13C–13C INADEQUATE/SARCOSY, 19F–13C HETCOR, and 1H–13C HETCOR, can be obtained at natural isotopic abundance within reasonable experiment times, thus enabling a complete resonance assignment of sitagliptin, a pharmaceutical used for the treatment of type 2 diabetes.

Measuring the 3J HNHa coupling by a simple 2D-intra-HNCA IP/AP-E.COSY with simultaneous encoding of 15N chemical shift and 1J HaCa evolution

The 3J coupling values are commonly used in biomolecular NMR to extract structural information. Here we present a novel intra HNCA IP/AP E.COSY pulse sequence that allows to measure 3J HNHa coupling constants by a simple and rapid two-dimensional 1H-15N correlation experiment where the 15N frequency is encoded at the same time as the 1J HaCa coupling evolution. The advantage with respect to the conventional 3D HNCA E.COSY pulse sequence is the dimensionality reduction to a simple 2D experiment, which decreases acquisition time and facilitates data analysis. The performance of this new experiment is demonstrated with an ubiquitin sample at 500 MHz.

Bulk and Surface Chemistry of the Niobium MAX and MXene Phases from Multinuclear Solid-State NMR Spectroscopy.

MXenes, derived from layered MAX phases, are a class of two-dimensional materials with emerging applications in energy storage, electronics, catalysis, and other fields due to their high surface areas, metallic conductivity, biocompatibility, and attractive optoelectronic properties. MXene properties are heavily influenced by their surface chemistry, but a detailed understanding of the surface functionalization is still lacking. Solid-state nuclear magnetic resonance (NMR) spectroscopy is sensitive to the interfacial chemistry, the phase purity including the presence of amorphous/nanocrystalline phases, and the electronic properties of the MXene and MAX phases. In this work, we systematically study the chemistry of Nb MAX and MXene phases, Nb2AlC, Nb4AlC3, Nb2CTx, and Nb4C3Tx, with their unique electronic and mechanical properties, using solid-state NMR spectroscopy to examine a variety of nuclei (1H, 13C, 19F, 27Al, and 93Nb) with a range of one- and two-dimensional correlation, wide-line, high-sensitivity, high-resolution, and/or relaxation-filtered experiments. Hydroxide and fluoride terminations are identified, found to be intimately mixed, and their chemical shifts are compared with other MXenes. This multinuclear NMR study demonstrates that diffraction alone is insufficient to characterize the phase composition of MAX and MXene samples as numerous amorphous or nanocrystalline phases are identified including NbC, AlO6 species, aluminum nitride or oxycarbide, AlF3·nH2O, Nb metal, and unreacted MAX phase. To the best of our knowledge, this is the first study to examine the transition-metal resonances directly in MXene samples, and the first 93Nb NMR of any MAX phase. The insights from this work are employed to enable the previously elusive assignment of the complex overlapping 47/49Ti NMR spectrum of Ti3AlC2. The results and methodology presented here provide fundamental insights on MAX and MXene phases and can be used to obtain a more complete picture of MAX and MXene chemistry, to prepare realistic structure models for computational screening, and to guide the analysis of property measurements.

Measurement of Accurate Interfluorine Distances in Crystalline Organic Solids: A High-Frequency Magic Angle Spinning NMR Approach.

Long-range interatomic distance restraints are critical for the determination of molecular structures by NMR spectroscopy, both in solution and in the solid state. Fluorine is a powerful NMR probe in a wide variety of contexts, owing to its favorable magnetic properties, ease of incorporation into biological molecules, and ubiquitous use in synthetic organic molecules designed for diverse applications. Because of the large gyromagnetic ratio of the 100% naturally abundant 19F isotope, interfluorine distances as long as 20 Å are accessible in magic-angle spinning (MAS) dipolar recoupling experiments. Herein, we present an approach for the determination of accurate interfluorine distances in multispin systems, using the finite pulse radio frequency driven recoupling (fpRFDR) at high MAS frequencies of 40-60 kHz. We use a series of crystalline "molecular ruler" solids, difluorobenzoic acids and 7F-L-tryptophan, for which the intra- and intermolecular interfluorine distances are known. We describe the optimal experimental conditions for accurate distance determinations, including the choice of a phase cycle, the relative advantages of selective inversion one-dimensional versus two-dimensional correlation experiments, and the appropriate numerical simulation protocols. An optimal strategy for the analysis of RFDR exchange curves in organic solids with extended spin interaction networks is presented, which, even in the absence of crystal structures, can be potentially incorporated into NMR structure determination.

Chemically Modified Silica Materials as Model Systems for the Characterization of Water-Surface Interactions

A series of novel functionalized mesoporous silica-based materials with well-defined pore diameters, surface functionalization and surface morphology is synthesized by co-condensation or grafting techniques and characterized by solid-state NMR spectroscopy, DNP enhanced solid state-NMR and thermodynamic techniques. These materials are employed as host-systems for small-guest molecules like water, small alcohols, carbonic acids, small aromatic molecules, binary mixtures and others. The phase-behavior of these confined guests is studied by combinations of one dimensional solid-state NMR techniques (¹HMAS, ²H-line shape analysis, ¹³C CPMAS) and two-dimensional correlation experiments like ¹H-²⁹Si- solid-state HETCOR.

13C/19F high-resolution solid-state NMR studies on layered carbon-fluorine compounds

13C/19F high-resolution solid-state NMR was applied to examine local structures of a stage-1 layered carbon-fluorine compound (C2.8F). Four 19F (F1∼F4) and two 13C signals (C1 and C2) unraveled by high magnetic field (14 T) and fast magic-angle spinning (>35 kHz) were examined by various two-dimensional correlation experiments. In addition to “through space” 13C-19F and 19F-19F dipolar correlation, which reveals distance proximity among 13C/19F spins, we examined feasibility of applying the J interaction for examination of “through bond” correlation. These experiments led assignment of two of the four F signals (F2 and F3) to F directly covalent bonded to sp3 carbon and an interleaving domain for the local structure of the minor C2-F3 group among the major domain composed of C2-F2 and sp2 carbon (C1). The other two 19F signals (F1 and F4) were assigned to as CF2 and F ions, respectively. A spectroscopic evidence for the C-F bond being the σ bond is given by the observation of a non-zero one-bond J value (193 ± 4 Hz) for C2-F. Further, the similar JCF = 197 Hz for C-F in poly(carbon fluoride) confirmed that the so-called “semi-ionic/semi-covalent” C-F bond in C2.8F is actually a “standard” covalent C-F bond.

Chemically Modified Silica Materials as Model Systems for the Characterization of Water-Surface Interactions

Abstract A series of novel functionalized mesoporous silica-based materials with well-defined pore diameters, surface functionalization and surface morphology is synthesized by co-condensation or grafting techniques and characterized by solid-state NMR spectroscopy, DNP enhanced solid state-NMR and thermodynamic techniques. These materials are employed as host-systems for small-guest molecules like water, small alcohols, carbonic acids, small aromatic molecules, binary mixtures and others. The phase-behavior of these confined guests is studied by combinations of one dimensional solid-state NMR techniques (1H MAS, 2H-line shape analysis, 13C CPMAS) and two-dimensional correlation experiments like 1H-29Si- solid-state HETCOR.

Polymorphs of Theophylline Characterized by DNP Enhanced Solid-State NMR.

We show how dynamic nuclear polarization (DNP) enhanced solid-state NMR spectroscopy can be used to characterize polymorphs and solvates of organic solids. We applied DNP to three polymorphs and one hydrated form of the asthma drug molecule theophylline. For some forms of theophylline, sample grinding and impregnation with the radical-containing solution, which are necessary to prepare the samples for DNP, were found to induce polymorphic transitions or desolvation between some forms. We present protocols for sample preparation for solid-state magic-angle spinning (MAS) DNP experiments that avoid the polymorphic phase transitions in theophylline. These protocols include cryogrinding, grinding under inert atmosphere, and the appropriate choice of the impregnating liquid. By applying these procedures, we subsequently demonstrate that two-dimensional correlation experiments, such as 1H–13C and 1H–15N HETCOR or 13C–13C INADEQUATE, can be obtained at natural isotopic abundance in reasonable times, thus enabling more advanced structural characterization of polymorphs.

Copper ESEEM and HYSCORE through ultra-wideband chirp EPR spectroscopy.

The main limitation of pulse electron paramagnetic resonance (EPR) spectroscopy is its narrow excitation bandwidth. Ultra-wideband (UWB) excitation with frequency-swept chirp pulses over several hundreds of megahertz overcomes this drawback. This allows to excite electron spin echo envelope modulation (ESEEM) from paramagnetic copper centers in crystals, whereas up to now, only ESEEM of ligand nuclei like protons or nitrogens at lower frequencies could be detected. ESEEM spectra are recorded as two-dimensional correlation experiments, since the full digitization of the electron spin echo provides an additional Fourier transform EPR dimension. Thus, UWB hyperfine-sublevel correlation experiments generate a novel three-dimensional EPR-correlated nuclear modulation spectrum.

1H and 2H NMR: Diagonal suppressed total correlation spectroscopy in spin echo mode

We report a novel diagonal suppressed two-dimensional total correlation experiment in solution state and demonstrate its performance in 1H and 2H NMR. In DIagonal Suppressed spin echo TOtal Correlation SpectroscopY (DISTOCSY), diagonal peaks which occur around F1=0 in spin echo mode are removed by conversion to unobservable multiple quantum or longitudinal terms in each scan of a two scan procedure, on application of an appropriately phased 90° pulse at the time of the echo maximum. The limitation of this approach to short FID acquisition times is discussed, and an alternative two-scan version of DISTOCSY for high resolution situations is presented.

Local Structure and Spin State of Cobalt Ion at Defect in Lithium Overstoichiometric LiCoO2 As Studied by 6/7Li Solid-State NMR Spectroscopy

Lithium overstoichiometric LixCoO2 (x > 1) with few phase transitions is promising as a positive electrode material for lithium ion batteries; however, the detailed lithium local structures around excess Li ions remain unknown and need to be elucidated. It has been postulated that the excess Li ion occupies the Co site, and the resulting charge deficit is compensated for by oxygen vacancies and the valence of Co. Two spin states have been invoked for the Co ion; one is Co3+ taking an intermediate spin state (S = 1) and the other is low spin Co2+ (S = 1/2). The paramagnetic shift caused by the Co spin is appreciable by 6/7Li nuclear magnetic resonance (NMR) in solids using magic-angle spinning (MAS), and we show that there are five “major” minor signals at ca. −16, −6, 3, 185, and 1100 ppm with the intensity ratio of 2:4:4:2:0.5–1.0, in addition to the major signal at 0 ppm. Analysis of temperature-dependent 7Li shifts and also 6Li T1 experiments confirm that the observed shifts for these minor peaks are ascribed to the paramagnetic interaction. To examine distance proximities among these lithium ions at the minor sites, we apply three 7Li–7Li two-dimensional (2D) correlation experiments and show that the three minor lithium atoms, whose shifts are 3, −6, and −16 ppm, exist in the same Li layer. Further, from rotational-resonance recoupling experiment, it is indicated that the lithium appearing at −16 ppm is closest to that at ca. 185 ppm. Based on these results, we show that a defect structure assuming low spin Co2+ which is not associated with the oxygen vacancy is consistent with the NMR results. This leads us to invoke Li insertion at an interstitial site, which has been negated, for the position of the excess Li atom.

Efficient band-selective homonuclear CO–CA cross-polarization in protonated proteins

Previously introduced for highly deuterated proteins, band-selective magnetization transfer between CO and CA spins by dipolar-based homonuclear cross polarization is applied here to a protonated protein. Robust and efficient recoupling is achieved when the sum of effective radio-frequency fields on CO and CA resonances equals two times the spinning rate, yielding up to 33% of magnetization transfer efficiency in protonated ubiquitin. The approach is designed for moderate magic-angle spinning rates and high external magnetic fields when the isotropic chemical shift difference of CO and CA considerably exceeds the spinning rate. This method has been implemented in NiCOi-1CAi-1 and CAi(Ni)COi-1CAi-1 two-dimensional interresidual correlation experiments for fast and efficient resonance assignment of ubiquitin by solid-state NMR spectroscopy.

Imaging of the B1 distribution and background signal in a MAS NMR probehead using inhomogeneous B0 and B1 fields

Several widely used methods for suppressing the “background” signal in 1H magic angle spinning (MAS) NMR spectroscopy are based on the assumption of a significant difference between the B1 radiofrequency field experienced by the sample (within the MAS rotor) and that felt by static components of the probehead (where the background signal is believed to originate). In this work, a two-dimensional correlation experiment employing inhomogeneous B0 and B1 fields is used to image the B1 distribution in a MAS NMR probehead. The experiment, which can be performed on any spectrometer, allows the distribution of the B1 field to be measured and also correlated with the spatial location of the NMR signal within the probehead. The method can also readily be combined with various “depth pulse” techniques for background suppression, allowing their performances to be more rigorously evaluated.

In-Cell NMR Spectroscopy–In vivo Monitoring of the Structure, Dynamics, Folding, and Interactions of Proteins at Atomic Resolution

Thallapuranam K Suresh Kumar, Ryan Thurman and Srinivas Jayanthi-In-Cell NMR Spectroscopy–<em>In vivo</em> Monitoring of the Structure, Dynamics, Folding, and Interactions of Proteins at Atomic Resolution

Preparation and Structural Characterization of Amorphous Spray-Dried Dispersions of Tenoxicam with Enhanced Dissolution

Tenoxicam is a poorly soluble nonsteroidal anti-inflammatory drug. In this work, the solubility of tenoxicam is enhanced using amorphous spray-dried dispersions (SDDs) prepared using two molar equivalents of l-arginine and optionally with 10%-50% (w/w) polyvinylpyrrolidone (PVP). When added to the dispersions, PVP is shown to improve physical properties and also assists in maintaining supersaturation in solution. The dispersions provide a twofold increase over equilibrium solubility at the same pH. The dispersions are characterized using electron microscopy, vibrational spectroscopy, diffuse-reflectance visible spectroscopy, and X-ray powder diffraction. The structures of the dispersions are probed using solid-state nuclear magnetic resonance (SSNMR) experiments applied to the (1) H, (13) C, and (15) N nuclei, including two-dimensional dipolar correlation experiments that detect molecular association and the formation of a glass solution between tenoxicam, l-arginine, and PVP. Other aspects of the amorphous structure, including hydrogen-bonding interactions and the ionization state of tenoxicam and l-arginine, are also explored using SSNMR methods. These methods are used to show that the SDDs contain an amorphous l-arginine salt of tenoxicam in a glass solution that also includes PVP when present. Finally, the dispersions show only a minor decrease in chemical stability during accelerated stability studies relative to a crystalline form of tenoxicam.

Disentangling Crystallographic Inequivalence and Crystallographic Forms of l-Arginine by One- and Two-Dimensional Solid-State NMR Spectroscopy

Overlapping (13)C or (15)N solid-state NMR spectra from crystallographically different forms of L-arginine hydrochloride can be separated by exploiting differential proton T(1) relaxation in conjunction with cross-polarization. Dipolar (13)C-(13)C and (15)N-(15)N two-dimensional correlation experiments reveal resonances belonging to crystallographically and magnetically inequivalent molecules.

Delineating pMDI model reactions with loblolly pine via solution-state NMR spectroscopy. Part 2. Non-catalyzed reactions with the wood cell wall

Abstract Solution-state NMR provides a powerful tool to observe the presence or absence of covalent bonds between wood and adhesives. Finely ground wood can be dissolved in an NMR-compatible solvent system containing dimethylsulfoxide-d 6 and N-methylimidazole-d 6 , in which the wood polymers remain largely intact. High-resolution solution-state two-dimensional NMR correlation experiments, based on 13C–1H one-bond heteronuclear single quantum coherence, allow structural analysis of the major cell wall components. This technique was applied to loblolly pine that was treated with polymeric methylene diphenyl diisocyanate (pMDI) related model compounds under controlled moisture and temperature conditions. Chemical shifts of carbamates formed between the pMDI model compounds and loblolly pine were determined. The results show that: (a) under dry conditions and a high concentration of isocyanate, carbamates will form preferentially with side-chain hydroxyl groups on β-aryl ether and phenylcoumaran-linked lignin units in a swelling solvent; (b) phenyl isocyanate is more capable of derivatization in the cell wall than the bulkier 4-benzylphenyl isocyanate; (c) at 5% and 14% moisture content, detectable carbamates on lignin side-chains dramatically decrease; and (d) under typical conditions of industrial oriented strand-board production in a hot press at 5% and 14% moisture content, no carbamate formation was detected.

Two-dimensional NMR correlation experiments in the gas phase

The application of common two-dimensional NMR correlation experiments to gaseous analytes for structural elucidation is reported. Standard sequences such as COSY, HSQC, and HMBC are readily applied to volatile hydrocarbons and fluorocarbons. In experiments using 19F or 13C as the observed nucleus, it is possible to take advantage of efficient spin-rotation relaxation to perform common experiments swiftly (a 19F COSY acquired in 6 s is shown) or to render insensitive experiments possible on a practical timescale (e.g. a gas phase INADEQUATE at natural isotopic abundance in 14 h). NOE-based experiments were not successful on the gaseous systems studied.