Homonuclear Coupling Research Articles

Clean pure shift 2D J‐resolved spectroscopy

2D J-resolved spectroscopy is one of the oldest and conceptually most elegant ways to separate homonuclear scalar couplings from chemical shift information. In practice, the classical experiment suffers from a number of complications that limits accuracy and resolution, including phasetwist lineshapes, strong coupling artifacts, and the need for shearing the spectrum by 45° to obtain a (J,δ)-representation. Here, a novel pure shift 2DJ experiment based on the TSE-PSYCHE experiment is reported that deals with all these issues. Previous experiments proposed z-filtration to avoid excessive artifacts caused by chunked pure shift acquisition. It will be shown that these artifacts can also easily be avoided by means of the Pell-Keeler method. As opposed to its z-filtered counterparts, the new experiment provides pure shift 2DJ spectra that are free of artifacts from pulse imperfections and minimize responses related to strong coupling. In this way, multiplet analysis becomes possible at maximal resolution and a minimum of spectral complications.

Analyses of Complex Mixtures by F1 Homo-Decoupled Diagonal Suppressed Total Correlation Spectroscopy.

A diagonal suppressed F1 decoupled total correlation spectroscopy(TOCSY) experiment is developed for analyses of complex mixtures. In 2D homonuclear correlation, assignment of the cross peaks is crucial for structure elucidation. However, when cross peaks are close to the diagonal peaks in overcrowded spectral regions, their assignment becomes tedious. In complex mixtures, the presence of multiple spectra along with broad and complex proton multiplets owing to homonuclear scalar couplings degrade the resolution to the extent that assignment of various cross peaks becomes tedious or impossible. Herein, a diagonal suppressed total correlation technique with F1 decoupling is presented to improve the resolution of the cross peaks. The resolution of the cross peaks is improved by both diagonal suppression as well as the collapse of the multiplets to singlets. Application of the method to a few mixtures of organic compounds reveals better identification of the cross peaks relative to other TOCSY variants.

Assignment of the absolute configuration of hepatoprotective highly oxygenated triterpenoids using X-ray, ECD, NMR J-based configurational analysis and HSQC overlay experiments

BackgroundThe plants of the genus Kadsura are widely distributed in China, South Korea, and Japan. Their roots and stems are traditionally used to treat blood diseases and pain. The main bioactive constituents of Kadsura longipedunculata comprise highly oxygenated triterpenoids. Schiartane-type nortriterpenoids showed anti-HIV, anti-HBV, and cytotoxic bioactivities. For such compounds, the absolute configuration influences the bioactivities, and hence its unambiguous determination is essential. In this work, the absolute configurations of three highly oxygenated schiartane-type nortriterpenoids were unequivocally assigned using X-ray, ECD, and J-based configuration analysis and HSQC overlay data. MethodsThe ethanol extract of Kadsura longipedunculata Finet et Gagnep was purified by column chromatography using silica, Sephadex LH-20, and ODS as substrates. To help assign the absolute configuration of schiartane-type nortriterpenoids, X-ray diffraction analysis, ECD experiment compared to ab initio computed data, DP4+ analysis, HSQC overlay, NOESY, and J-based configuration analysis were carried out. Hetero- and homo-nuclear coupling constants were extracted from HETLOC experiments. ResultsThree new highly oxygenated triterpenoids, micrandilactone I (1), micrandilactone J (2), and 22,23-di-epi-micrandilactone J (3) were isolated. Their 2D structures were solved using NMR and HRESIMS data and their absolute configurations were elucidated using X-ray diffraction analysis, ECD experimental results compared to ab initio computed spectra, HSQC overlay, DP4+, NOESY, and J-based configuration analysis. Micrandilactone I (1) and 22,23-di-epi-micrandilactone J (3) showed moderate hepatoprotective activity against APAP-induced toxicity in HepG2 cells with cell survival rates of 53.0 and 50.2%, respectively, at 10μM (bicyclol, 49.0%), while micrandilactone J (2) was inactive. General significanceThis is the first comprehensive stereochemical assignment of a non-crystalline schiartane-type nortriterpenoid like 3. This general protocol may contribute towards solving the problems hampering the assignment of the absolute configurations of other members of this class of nortriterpenoids.

Nuclear magnetic resonance surface relaxation mechanisms of kerogen

Nuclear magnetic resonance (NMR) relaxometry is an excellent tool for probing the interactions between solid pore surface and pore fluids in porous media. Surface relaxation is a key component of NMR relaxation. It is well-known that in conventional rocks, paramagnetic centers contribute most to the surface relaxation phenomenon. However, the interactions between organic pore surfaces and pore fluids, and the mechanism of surface relaxation in organic shale pores, are not well-understood. We tackle the issue using deuterated compounds to adjust the proton density in the liquid phase and monitoring the transverse relaxation rate changes of kerogen-fluid mixtures. With the Barnett and Eagle Ford kerogen isolates, we found that for alkanes, it is intramolecular dipolar coupling that dominates among the magnetic interactions. As a result, the transverse relaxation rate of alkane proton spins is more likely to be dependent on the concentration of active adsorption sites on the kerogen surface, rather than the kerogen proton density. For water inside organic pores, surface relaxation most likely originates from hydrogen bonding and intermolecular dipolar coupling. We also examined the temperature effect on kerogen surface relaxation and found temperature-dependent behavior that is consistent with surface relaxation by hydrogen bonding and homonuclear dipolar coupling interactions.

SHARPER Reaction Monitoring: Generation of a Narrow Linewidth NMR Singlet, without X-Pulses, in an Inhomogeneous Magnetic Field.

We report a new pure-shift method, termed SHARPER (Sensitive, Homogeneous, And Resolved PEaks in Real time) designed for the analysis of reactions and equilibria by NMR. By focusing on a single selected signal, SHARPER removes all heteronuclear couplings of a selected nucleus without the need to pulse on X channels, thus overcoming hardware limitations of conventional spectrometers. A more versatile decoupling scheme, termed sel-SHARPER, removes all heteronuclear and homonuclear couplings of the selected signal. Both methods are characterized by a periodic inversion of the active spin during the real-time acquisition. In addition to decoupling, they also compensate for pulse imperfections and magnetic field inhomogeneity, generating an extremely narrow singlet with a linewidth approaching limits dictated by the spin–spin relaxation. The decoupling and line narrowing effected by (sel)-SHARPER provide significant increases in the signal-to-noise (S/N) ratio. Increases of 20-fold were routinely achieved for 19F detection. sel-SHARPER is also applicable to first- and higher-order 1H spectra. The sensitivity gains are substantially greater for inhomogeneous magnetic fields, including dynamic inhomogeneity caused by gas sparging. The parameters of the pulse sequences have been analyzed in detail to provide guidelines for their most effective application. The considerable reduction in the detection threshold induced by (sel)-SHARPER make the technique particularly suited for in situ monitoring of reaction kinetics. The approach is illustrated by a 19F NMR study of the protodeboronation of an aryl boronic acid. Here, the high S/N allowed reliable determination of the net protodeoboronation kinetics, and the excess line broadening of 19F singlets was utilized to characterize the boronic acid/boronate equilibrium kinetics. Oxidation of diphenylphosphine, monitored by 31P NMR under optimized gas-flow conditions, demonstrated the high tolerance of SHARPER to dynamic inhomogeneity. The principles of the (sel)-SHARPER sequences are expected to find numerous applications in the design of new NMR experiments.

Highly resolved HSQC experiments for the fast and accurate measurement of homonuclear and heteronuclear coupling constants

A number of J-upscaled NMR experiments are currently available to measure coupling constants along the indirect F1 dimension of a 2D spectrum. A major drawback is the limited F1 digital resolution that requires long acquisition times in order to achieve reasonably accurate measures. Here is shown how high levels of F1 digital resolution in a multiple-purpose HSQC experiment can be easily achieved by implementing a general J/δ-scaling strategy. In particular, a set of new J-resolved HSQC experiments is presented for a faster and much more accurate J determination in small molecules. Several options and practical aspects are discussed and exemplified by measuring the magnitude and/or the sign of several homo- and heteronuclear coupling constants in one shot.

Magic Echo for Nuclear Magnetic Resonance Characterization of Shales

Low-field nuclear magnetic resonance (NMR) has been widely applied in the oil industry as a noninvasive technique for rock analysis. There are many organic compounds, such as kerogen, in unconventional shale resources. However, it is difficult to evaluate solid organic matter in shales using common NMR techniques. Previous work has shown the use of a solid echo on shale samples provides more information than the standard T1–T2 method with spin echo. But for multiple homonuclear dipolar coupling present in shales, solid echo is less efficient. We propose a new multimagic-echo sequence for T1 and T1–T2 measurements. The magic echo and standard T1 and T2 measurement technologies are combined to provide a more complete evaluation of hydrogen-containing shale sample components. Compared with other measurements, magic-echo measurements provide more signal of organic matters in shale samples. The additional signal is contributed by homonuclear dipolar coupling in organic matter. The partial least-squares regress...

Significance of symmetry in the nuclear spin Hamiltonian for efficient heteronuclear dipolar decoupling in solid-state NMR: A Floquet description of supercycled rCW schemes.

Symmetry plays an important role in the retention or annihilation of a desired interaction Hamiltonian in NMR experiments. Here, we explore the role of symmetry in the radio-frequency interaction frame Hamiltonian of the refocused-continuous-wave (rCW) pulse scheme that leads to efficient 1H heteronuclear decoupling in solid-state NMR. It is demonstrated that anti-periodic symmetry of single-spin operators (Ix, Iy, Iz) in the interaction frame can lead to complete annihilation of the 1H-1H homonuclear dipolar coupling effects that induce line broadening in solid-state NMR experiments. This symmetry also plays a critical role in cancelling or minimizing the effect of 1H chemical-shift anisotropy in the effective Hamiltonian. An analytical description based on Floquet theory is presented here along with experimental evidences to understand the decoupling efficiency of supercycled (concatenated) rCW scheme.

Detection of intermolecular homonuclear dipolar coupling in organic rich shale by transverse relaxation exchange

The mechanism behind surface relaxivity within organic porosity in shales has been an unanswered question. Here, we present results that confirm the existence of intermolecular homonuclear dipolar coupling between solid and liquid phases in sedimentary organic matter. Transverse magnetization exchange measurements were performed on an organic-rich shale saturated with liquid hydrocarbon. Liquid and solid constituents were identified through both sample resaturation and through their T1/T2 ratios. Extensive cross peaks are observed in the T2-T2 exchange spectra between the solid and liquid constituents, indicating an exchange of magnetization between the two phases. This result cannot arise from physical molecular diffusion, and the dissolution energies are too high for chemical exchange, such that the magnetization exchange must arise from intermolecular homonuclear dipolar coupling. These results both confirm a possible source of surface relaxivity in organic matter and emphasize caution in the use of standard porous media interpretations of relaxation results in shales because of coupling between different magnetization environments.

Hydrogen incorporation mechanisms in forsterite: New insights from1H and29Si NMR spectroscopy and first-principles calculation

The presence of water (hydrogen) in nominally anhydrous mantle minerals may have profound effects on their physical properties (e.g., electrical conductivity, diffusivity, rheology), and these effects are expected to depend on how water is incorporated in the crystal structure. For olivine, the most abundant upper mantle mineral, despite extensive studies, mostly using vibrational spectroscopy, the interpretations are still not well constrained. To provide better understanding on this issue, we carried out a comprehensive 1 H and 29 Si NMR study on an Mg 2 SiO 4 forsterite sample containing about 0.5 wt% H 2 O synthesized at 12 GPa and 1200 °C, complemented by Raman measurement and first-principles calculation of the geometry, stability, and NMR parameters of model structures. The Raman spectra contain relatively sharp O-H stretching bands near 3612, 3579, and 3567 cm −1 and a broader band near 3547 cm −1 , similar to previous reports. The 1 H static and MAS NMR data revealed that there are two main populations of protons in the hydrous forsterite structure, one experiencing strong 1 H- 1 H homonuclear dipolar couplings and contributing to a broad peak near 2.4 ppm, and another with weaker dipolar couplings and contributing to a narrower peak near 1.2 ppm in the MAS NMR spectrum at 30 kHz. Two-dimensional 1 H CRAMPS-MAS NMR measurements confirmed that the two proton components belong to the same phase and the contrast in MAS NMR peak width is largely due to difference in the strength of 1 H- 1 H homonuclear dipolar couplings. In addition, there is also a very weak, narrow 1 H MAS NMR peak near 7.3 ppm (contributing to Si ] in a tetrahedral site of olivine, but unlike hydrogarnet with one of the protons pointing away from the tetrahedral center and located in an adjacent interstitial site, thus experiencing weaker dipolar couplings than those in the vicinity of the vacant tetrahedron; the very weak narrow peak near 7.3 ppm can be attributed to the substitution mechanism of two H ions for one Mg in an M1 site [(2H) M1 ] of forsterite. The 1 H- 29 Si CP-MAS NMR spectra revealed both a broad peak encompassing the position for OH defect-free forsterite (−61.7 ppm) and a narrower peak at higher frequency (−60.9 ppm). First-principles calculation indicates that these peaks are accountable by the same models as for the 1 H NMR data. Thus, this study has provided unambiguous evidence supporting that hydrogen is incorporated in forsterite at relatively high-pressure dominantly as (4H) Si defects, with (2H) M1 defects playing only a very minor role. The much larger 1 H chemical shift for protons associated with the latter (than the former) is correlated with stronger hydrogen bonding for the latter, which in turn reflects difference in bonding environments of the OH groups (with the latter bonded to a Si, and the former only bonded to Mg). Similar correlation applies to the O-H stretching frequency. The (4H) Si defects are responsible for the observed high-frequency O-H stretching bands (>3450 cm −1 ), and the (2H) M1 defects give lower frequencies (undetected here due to low abundance, but most likely near 3160–3220 cm −1 as previously reported) in vibrational spectra. These results can serve as a guide for (re-)interpretation of infrared and Raman spectroscopic data on hydrous olivine produced under different pressure and silica activity conditions, and require reconsideration of any models for the effects of water on physical properties of olivine based on different interpretations of such data. This study also demonstrated the usefulness of the combined solid-state NMR and first-principles calculation approach in unraveling the hydrogen incorporation mechanisms in nominally anhydrous minerals.

Anatomising proton NMR spectra with pure shift 2D J-spectroscopy: A cautionary tale

Analysis of proton NMR spectra has been a key tool in structure determination for over 60years. A classic tool is 2D J-spectroscopy, but common problems are the difficulty of obtaining the absorption mode lineshapes needed for accurate results, and the need for a 45° shear of the final 2D spectrum. A novel 2D NMR method is reported here that allows straightforward determination of homonuclear couplings, using a modified version of the PSYCHE method to suppress couplings in the direct dimension. The method illustrates the need for care when combining pure shift data acquisition with multiple pulse methods.

S3 HMBC hetero: Spin-State-Selective HMBC for accurate measurement of long-range heteronuclear coupling constants

A novel method, Spin-State-Selective (S3) HMBC hetero, for accurate measurement of heteronuclear coupling constants is introduced. The method extends the S3 HMBC technique for measurement of homonuclear coupling constants by appending a pulse sequence element that interchanges the polarization in 13C-1H methine pairs. This amounts to converting the spin-state selectivity from 1H spin states to 13C spin states in the spectra of long-range coupled 1H spins, allowing convenient measurement of heteronuclear coupling constants similar to other S3 or E.COSY-type methods. As usual in this type of techniques, the accuracy of coupling constant measurement is independent of the size of the coupling constant of interest. The merits of the new method are demonstrated by application to vinyl acetate, the alkaloid strychnine, and the carbohydrate methyl β-maltoside.

1H–1H Double Quantum NMR Investigation of Proton Dynamics in Solid Acids

Currently, the most popular proton exchange membrane (PEM) for fuel cell applications is Nafion. However, Nafion does not retain its high conductivity at high temperatures due to its dependence on water for proton transport. Because operational temperatures higher than the evaporation point of water are desirable, a family of solid acids was investigated. Cations known to transport protons were paired with anions to make acidic salts. Solid acids discussed here include imidazole paired with trifluoromethanesulfuric acid as well as imidazole, benzimidazole and adenine paired with methanesulfonic acid. Solid-state NMR was utilized to show the relative mobility of protons through double-quantum filter (DQF) experiments. The POST-C7 homonuclear dipolar-recoupling scheme was paired with DUMBO homonuclear decoupling to produce 1H double-quantum coherence buildup curves for the hydrogen-bonded protons of interest. Experimental buildup curves, which reflect both local structure as well as dynamics, are compared to theoretical curves of the static system. The SPINEVOLUTION-simulated curves utilized up to eight pairs of homonuclear dipolar couplings within a sphere of 7 Å diameter centered on the proton of interest. Steep buildup of the DQ curve and maxima at short recoupling times in the buildup curves indicate strong dipole–dipole coupling and are interpreted to indicate limited dynamics of the H-bonded protons. In contrast, shallower buildup curves and maxima at longer recoupling times imply that H-bonded protons (in an otherwise similar structure) are associated with local mobility, which reduces their local dipolar coupling and may facilitate proton transport. Bulk proton conductivities measured via electrochemical impedance spectroscopy were compared to DQF measurements to understand proton conduction within these materials.

Exciting Wide NMR Spectra of Static Solid Samples with Weak Radiofrequency Fields

Abstract Trains of short pulses in the manner of ‘delays alternating with nutations for tailored excitation’ (DANTE) have been applied to the Pake patterns of protons of water molecules trapped in a static powdered sample of barium chlorate monohydrate. The spin dynamics in the course of such experiments have been investigated by means of numerical simulations and compared with the ideal refocusing that can be achieved under magic-angle spinning (MAS). Solid echoes yield essentially undistorted lineshapes, in contrast to direct excitation without refocusing that leads to severe dispersions of the phases because of inhomogeneous interactions such as homonuclear dipolar couplings and anisotropic chemical shifts. The selectivity of DANTE sequences allows one to access ‘slices’ of the Pake pattern that can be related to particular crystallite orientations. Single-crystal spectra can therefore be extracted from powder spectra. A similar behavior is expected for both dipolar and quadrupolar echoes.

Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR Spectroscopy.

In this work an improved stable isotope labeling protocol for nucleic acids is introduced. The novel building blocks eliminate/minimize homonuclear 13C and 1H scalar couplings thus allowing proton relaxation dispersion (RD) experiments to report accurately on the chemical exchange of nucleic acids. Using site‐specific 2H and 13C labeling, spin topologies are introduced into DNA and RNA that make 1H relaxation dispersion experiments applicable in a straightforward manner. The novel RNA/DNA building blocks were successfully incorporated into two nucleic acids. The A‐site RNA was previously shown to undergo a two site exchange process in the micro‐ to millisecond time regime. Using proton relaxation dispersion experiments the exchange parameters determined earlier could be recapitulated, thus validating the proposed approach. We further investigated the dynamics of the cTAR DNA, a DNA transcript that is involved in the viral replication cycle of HIV‐1. Again, an exchange process could be characterized and quantified. This shows the general applicablility of the novel labeling scheme for 1H RD experiments of nucleic acids.

Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR Spectroscopy

AbstractIn this work an improved stable isotope labeling protocol for nucleic acids is introduced. The novel building blocks eliminate/minimize homonuclear 13C and 1H scalar couplings thus allowing proton relaxation dispersion (RD) experiments to report accurately on the chemical exchange of nucleic acids. Using site‐specific 2H and 13C labeling, spin topologies are introduced into DNA and RNA that make 1H relaxation dispersion experiments applicable in a straightforward manner. The novel RNA/DNA building blocks were successfully incorporated into two nucleic acids. The A‐site RNA was previously shown to undergo a two site exchange process in the micro‐ to millisecond time regime. Using proton relaxation dispersion experiments the exchange parameters determined earlier could be recapitulated, thus validating the proposed approach. We further investigated the dynamics of the cTAR DNA, a DNA transcript that is involved in the viral replication cycle of HIV‐1. Again, an exchange process could be characterized and quantified. This shows the general applicablility of the novel labeling scheme for 1H RD experiments of nucleic acids.

EXtended ACquisition Time (EXACT) NMR-A Case for 'Burst' Non-Uniform Sampling.

A strong case exists for the introduction of burst non-uniform sampling (NUS) in the direct dimension of NMR spectroscopy experiments. The resulting gaps in the NMR free induction decay can reduce the power demands of long experiments (by switching off broadband decoupling for example) and/or be used to introduce additional pulses (to refocus homonuclear coupling, for example). The final EXtended ACquisition Time (EXACT) spectra are accessed by algorithmic reconstruction of the missing data points and can provide higher resolution in the direct dimension than is achievable with existing non-NUS methods.

19 F DOSY diffusion-NMR spectroscopy of fluoropolymers.

A new pulse sequence for obtaining 19 F detected DOSY (diffusion ordered spectroscopy) spectra of fluorinated molecules is presented and used to study fluoropolymers based on vinylidene fluoride and chlorotrifluoroethylene. The performance of 19 F DOSY NMR experiments (and in general any type of NMR experiment) on fluoropolymers creates some unique complications that very often prevent detection of important signals. Factors that create these complications include: (1) the presence of many scalar couplings among 1 H, 19 F and 13 C; (2) the large magnitudes of many 19 F homonuclear couplings (especially 2 JFF ); (3) the large 19 F chemical shift range; and (4) the low solubility of these materials (which requires that experiments be performed at high temperatures). A systematic study of the various methods for collecting DOSY NMR data, and the adaptation of these methods to obtain 19 F detected DOSY data, has been performed using a mixture of low molecular weight, fluorinated model compounds. The best pulse sequences and optimal experimental conditions have been determined for obtaining 19 F DOSY spectra. The optimum pulse sequences for acquiring 19 F DOSY NMR data have been determined for various circumstances taking into account the spectral dispersion, number and magnitude of couplings present, and experimental temperature. Pulse sequences and experimental parameters for optimizing these experiments for the study of fluoropolymers have been studied. Copyright © 2016 John Wiley & Sons, Ltd.

Determination of unresolved heteronuclear scalar coupling constants by J(up)-HSQMBC

Long-range heteronuclear scalar coupling constants provide important structural information, which is necessary for obtaining stereospecific assignment or dihedral angle information. The measurement of small proton–carbon splittings is particularly difficult due to the low natural abundance of carbon-13 and the presence of homonuclear couplings of similar size. Here we present a real-time J-upscaled HSQMBC, which allows the measurement of heteronuclear coupling constants even if they are hidden in the signal linewidth of a regular spectrum.

Observation of potentially troublesome (2) JCC correlations in 1,1-ADEQUATE spectra.

Despite the tremendous usage of HMBC to establish long-range (1) H-(13) C and (1) H-(15) N heteronuclear correlations, an inherent drawback of the experiment is the indeterminate nature of the (n) JXH correlations afforded by the experiment. A priori there is no reliable way of determining whether a given (n) JCH correlation is, for example, via two-, three-, or sometimes even four-bonds. This limitation of the HMBC experiment spurred the development of the ADEQUATE family of NMR experiments that rely on, in the case of 1,1-ADEQUATE, an out-and-back transfer of magnetization via the (1) JCC homonuclear coupling constant, which is significantly larger than (n) JCC (where n = 2-4) couplings in most cases. Hence, the 1,1-ADEQUATE experiment has generally been assumed to unequivocally provide the equivalent of (2) JCH correlations. The recent development of the 1,1- and 1,n-HD-ADEQUATE experiments that can provide homodecoupling for certain (1) JCC and (n) JCC correlations has increased the sensitivity of the ADEQUATE experiments significantly and can allow acquisition of these data in a fraction of the time required for the original iterations of this pulse sequence. With these gains in sensitivity, however, there occasionally come unanticipated consequences. We have observed that the collapse of proton multiplets, in addition to providing better s/n for the desired (1) JCC correlations can facilitate the observation of typically weaker (2) JCC correlations across intervening carbonyl resonances in 1,1-HD-ADEQUATE spectra. Several examples are shown, with the results supported by the measurement of the (2) JCC coupling constants in question using J-modulated-HD-ADEQUATE and DFT calculations. Copyright © 2016 John Wiley & Sons, Ltd.