Solid-state NMR investigations of Si-29 and N-15 enriched silicon nitride

We compare ²⁹Si magic-angle spinning (MAS) nuclear magnetic resonance (NMR) spectra from the two modifications of silicon nitride, α-Si₃N₄ and β-Si₃N₄, with that of a fully (²⁹Si, ¹⁵N)-enriched sample ²⁹Si₃¹⁵N₄, as well as ¹⁵N NMR spectra of Si₃¹⁵N₄ (having ²⁹Si at natural abundance) and ²⁹Si₃¹⁵N₄. We show that the ¹⁵N NMR peak-widths from the latter are dominated by J(²⁹Si–¹⁵N) through-bond interactions, leading to significantly broader NMR signals compared to those of Si₃¹⁵N₄. By fitting calculated ²⁹Si NMR spectra to experimental ones, we obtained an estimated coupling constant J(²⁹Si–¹⁵N) of 20 Hz. We provide ²⁹Si spin-lattice (T₁) relaxation data for the ²⁹Si₃¹⁵N₄ sample and chemical shift anisotropy results for the ²⁹Si site of β-Si₃N₄. Various factors potentially contributing to the ²⁹Si and ¹⁵N NMR peak-widths of the various silicon nitride specimens are discussed. We also provide powder X-ray diffraction (XRD) and mass spectrometry data of the samples.     

C, N CPMAS NMR and GIAO DFT calculations of stereoisomeric oxindole alkaloids from Cat's Claw (Uncaria tomentosa)

Oxindole alkaloids, isolated from the bark of Uncaria tomentosa [Willd. ex Schult.] Rubiaceae, are considered to be responsible for the biological activity of this herb. Five pentacyclic and two tetracyclic alkaloids were studied by solid-state NMR and theoretical GIAO DFT methods. The ¹³C and ¹⁵N CPMAS NMR spectra were recorded for mitraphylline, isomitraphylline, pteropodine (uncarine C), isopteropodine (uncarine E), speciophylline (uncarine D), rhynchophylline and isorhynchophylline. Theoretical GIAO DFT calculations of shielding constants provide arguments for identification of asymmetric centers and proper assignment of NMR spectra. These alkaloids are 7R/7S and 20R/20S stereoisomeric pairs. Based on the ¹³C CP MAS chemical shifts the 7S alkaloids (δ C3 70–71 ppm) can be easily and conveniently distinguished from 7R (δC3 74.5–74.9 ppm), also 20R (δC20 41.3–41.7 ppm) from the 20S (δC20 36.3–38.3 ppm). The epiallo-type isomer (3R, 20S) of speciophylline is characterized by a larger ¹⁵N MAS chemical shift of N4 (64.6 ppm) than the allo-type (3S, 20S) of isopteropodine (δN4 53.3 ppm). ¹⁵N MAS chemical shifts of N1–H in pentacyclic alkaloids are within 131.9–140.4 ppm.     

Design and construction of a versatile dual volume heteronuclear double resonance microcoil NMR probe

Improved NMR detection of mass limited samples can be obtained by taking advantage of the mass sensitivity of microcoil NMR, while throughput issues can be addressed using multiple, parallel sample detection coils. We present the design and construction of a double resonance 300-MHz dual volume microcoil NMR probe with thermally etched 440-nL detection volumes and fused silica transfer lines for high-throughput stopped-flow or flow-through sample analysis. Two orthogonal solenoidal detection coils and the novel use of shielded inductors allowed the construction of a probe with negligible radio-frequency cross talk. The probe was resonated at ¹H–²D (upper coil) and ¹H–¹³C (lower coil) frequencies such that it could perform 1D and 2D experiments with active locking frequency. The coils exhibited line widths of 0.8–1.1 Hz with good mass sensitivity for both ¹H and ¹³C NMR detection. ¹³C-directly detected ²D HETCOR spectra of 5% v/v ¹³C labeled acetic acid were obtained in less than 5 min. Demonstration of the probe characteristics as well as applications of the versatile two-coil double resonance probe are discussed.     

Improved quantification of alite and belite in anhydrous Portland cements by Si MAS NMR: Effects of paramagnetic ions

The applicability, reliability, and repeatability of ²⁹Si MAS NMR for determination of the quantities of alite (Ca₃SiO₅) and belite (Ca₂SiO₄) in anhydrous Portland cement was investigated in detail for 11 commercial Portland cements and the results compared with phase quantifications based on powder X-ray diffraction combined with Rietveld analysis and with Taylor–Bogue calculations. The effects from paramagnetic ions (Fe³⁺) on the spinning sideband intensities, originating from dipolar couplings between ²⁹Si and the spins of the paramagnetic electrons, were considered and analyzed in spectra recorded at four magnetic fields (4.7–14.1 T) and this has led to an improved quantification of alite and belite from ²⁹Si MAS NMR spectra recorded at “high” spinning speeds of ν_R=12.0–13.0 kHz using 4 or 5 mm rotors. Furthermore, the impact of Fe³⁺ ions on the spin-lattice relaxation was studied by inversion-recovery experiments and it was found that the relaxation is overwhelmingly dominated by the Fe³⁺ ions incorporated as guest-ions in alite and belite rather than the Fe³⁺ sites present in the intimately mixed ferrite phase (Ca₂Al_xFe_2−xO₅).     

The 2D {P} Spin-Echo-Difference Constant-Time [C, H]-HMQC Experiment for Simultaneous Determination of JH3′P and JC4′P in C-Labeled Nucleic Acids and Their Protein Complexes

A two-dimensional {³¹P} spin-echo-difference constant-time [¹³C, ¹H]-HMQC experiment (2D {³¹P}-sedct-[¹³C, ¹H]-HMQC) is introduced for measurements of ³J_C4′P and ³J_H3′P scalar couplings in large ¹³C-labeled nucleic acids and in DNA–protein complexes. This experiment makes use of the fact that ¹H–¹³C multiple-quantum coherences in macromolecules relax more slowly than the corresponding ¹³C single-quantum coherences. ³J_C4′P and ³J_H3′P are related via Karplus-type functions with the phosphodiester torsion angles β and ε, respectively, and their experimental assessment therefore contributes to further improved quality of NMR solution structures. Data are presented for a uniformly ¹³C, ¹⁵N-labeled 14-base-pair DNA duplex, both free in solution and in a 17-kDa protein–DNA complex.     

Al and Si MAS NMR investigations of cordierite glass, μ- and α-cordierite

²⁷Al and ²⁹Si Magic-Angle Spinning NMR results are reported for conventionally prepared glass of cordierite stoichiometry (2MgO · 2Al₂O₃ · 5SiO₂), the metastable high-quartz solid solution (μ-cordierite) and the high-temperature polymorph of cordierite (α-cordierite). Both, ²⁷Al two-dimensional (2D) quadrupole nutation experiments and ²⁷Al satellite transition spectroscopy (SATRAS) have been applied to identify two different tetrahedrally-coordinated aluminium sites (AlO₄). SATRAS has been used to extract the quadrupole interaction parameters and their distribution, the isotropic chemical shifts and the relative populations of the different Al sites. Both, the ²⁷Al and ²⁹Si NMR results, lead to the conclusion that a perfect Si/Al disorder does not exist in these investigated cordierite samples.     

Ab initio predictions of zeolite structures and Si NMR chemical shifts

A recent shell-model potential parameterized on ab initio data is used for predicting the all-silica structures of zeolites MFI, MEI, MTW, TON, FAU and of α-quartz. Cluster models are defined around each site and the ²⁹Si NMR shielding constants are calculated by ab initio techniques (GIAO-HF). Good agreement with observed ²⁹Si NMR chemical shifts is found. Comparison is made with shifts calculated for observed structures. The structures predicted by the ab initio shell-model potential prove as accurate as the observed ones when judged on the quality of the calculated ²⁹Si NMR spectra.     

3D 1H-13C-14N correlation solid-state NMR spectrum

Nitrogen-14 (spin I = 1) has always been a nucleus difficult to observe in solid-state NMR and until recently its observation was restricted to one-dimensional (1D) spectra. We present here the first 3D ¹H–¹³C–¹⁴N NMR correlation spectrum. This spectrum was acquired on a test sample l-histidine·HCl·H₂O using a recently developed technique, which consists in indirectly observing ¹⁴N nuclei via dipolar recoupling with an HMQC-type experiment.     

Angular momentum dependence of the giant dipole resonance width in excited nuclei of mass

Gamma ray spectra in the energy range of 4–25 MeV were measured in the reaction ²⁸Si + ¹²⁴Sn at E(²⁸Si) 150 MeV in coincidence with low energy γ-multiplicities. The spectra were analysed using a simulated Monte Carlo CASCADE code. The centroid energy and width of the giant dipole resonance were extracted for various multiplicity windows. The average angular momentum and temperature of the final states populated after the giant dipole photon emission range from 25 and 1.5 MeV to 56 and 1.3 MeV, respectively. The extracted widths are almost constant for the lower multiplicity windows and show an increase of 1.4 MeV at the highest window. The nuclei are expected to be near the liquid drop regime and the experimental results are not inconsistent with the liquid drop behaviour.     

One- and two-dimensional C–H/N–H dipolar correlation experiments under fast magic-angle spinning for determining the peptide dihedral angle φ

A recently proposed ¹³C–¹H recoupling sequence operative under fast magic-angle spinning (MAS) [K. Takegoshi, T. Terao, Solid State Nucl. Magn. Reson. 13 (1999) 203–212.] is applied to observe ¹³C–¹H and ¹⁵N–¹H dipolar powder patterns in the ¹H–¹⁵N–¹³C–¹H system of a peptide bond. Both patterns are correlated by ¹⁵N-to-¹³C cross polarization to observe one- or two-dimensional (1D or 2D) correlation spectra, which can be simulated by using a simple analytical expression to determine the H–N–C–H dihedral angle. The 1D and 2D experiments were applied to N-acetyl[1,2-¹³C,¹⁵N] -valine, and the peptide φ angle was determined with high precision by the 2D experiment to be ±155.0°±1.2°. The positive one is in good agreement with the X-ray value of 154°±5°. The 1D experiment provided the value of φ=±156.0°±0.8°.     

CHHC and H–H magnetization exchange: Analysis by experimental solid-state NMR and 11-spin density-matrix simulations

A protocol is presented for correcting the effect of non-specific cross-polarization in CHHC solid-state MAS NMR experiments, thus allowing the recovery of the ¹H–¹H magnetization exchange functions from the mixing-time dependent buildup of experimental CHHC peak intensity. The presented protocol also incorporates a scaling procedure to take into account the effect of multiplicity of a CH₂ or CH₃ moiety. Experimental CHHC buildup curves are presented for l-tyrosine·HCl samples where either all or only one in 10 molecules are U–¹³C labeled. Good agreement between experiment and 11-spin SPINEVOLUTION simulation (including only isotropic ¹H chemical shifts) is demonstrated for the initial buildup (t_mix < 100 μs) of CHHC peak intensity corresponding to an intramolecular close (2.5 Å) H–H proximity. Differences in the initial CHHC buildup are observed between the one in 10 dilute and 100% samples for cases where there is a close intermolecular H–H proximity in addition to a close intramolecular H–H proximity. For the dilute sample, CHHC cross-peak intensities tended to significantly lower values for long mixing times (500 μs) as compared to the 100% sample. This difference is explained as being due to the dependence of the limiting total magnetization on the ratio N_obs/N_tot between the number of protons that are directly attached to a ¹³C nucleus and hence contribute significantly to the observed ¹³C CHHC NMR signal, and the total number of ¹H spins into the system. ¹H–¹H magnetization exchange curves extracted from CHHC spectra for the 100% l-tyrosine·HCl sample exhibit a clear sensitivity to the root sum squared dipolar coupling, with fast buildup being observed for the shortest intramolecular distances (2.5 Å) and slower, yet observable buildup for the longer intermolecular distances (up to 5 Å).     

Molecular dynamics and information on possible sites of interaction of intramyocellular metabolites in vivo from resolved dipolar couplings in localized H NMR spectra

Proton NMR resonances of the endogenous metabolites creatine and phosphocreatine ((P)Cr), taurine (Tau), and carnosine (Cs, β-alanyl-l-histidine) were studied with regard to residual dipolar couplings and molecular mobility. We present an analysis of the direct ¹H–¹H interaction that provides information on motional reorientation of subgroups in these molecules in vivo. For this purpose, localized ¹H NMR experiments were performed on m. gastrocnemius of healthy volunteers using a 1.5-T clinical whole-body MR scanner. We evaluated the observable dipolar coupling strength SD₀ (S = order parameter) of the (P)Cr-methyl triplet and the Tau-methylene doublet by means of the apparent line splitting. These were compared to the dipolar coupling strength of the (P)Cr-methylene doublet. In contrast to the aliphatic protons of (P)Cr and Tau, the aromatic H2 (δ = 8 ppm) and H4 (δ = 7 ppm) protons of the imidazole ring of Cs exhibit second-order spectra at 1.5 T. This effect is the consequence of incomplete transition from Zeeman to Paschen-Back regime and allows a determination of SD₀ from H2 and H4 of Cs as an alternative to evaluating the multiplet splitting which can be measured directly in high-resolution ¹H NMR spectra. Experimental data showed striking differences in the mobility of the metabolites when the dipolar coupling constant D₀ (calculated with the internuclear distance known from molecular geometry in the case of complete absence of molecular dynamics and motion) is used for comparison. The aliphatic signals involve very small order parameters S ≈ (1.4 − 3) × 10⁻⁴ indicating rapid reorientation of the corresponding subgroups in these metabolites. In contrast, analysis of the Cs resonances yielded S ≈ (113 − 137) × 10⁻⁴. Thus, the immobilization of the Cs imidazole ring owing to an anisotropic cellular substructure in human m. gastrocnemius is much more effective than for (P)Cr and Tau subgroups. Furthermore, ¹H NMR experiments on aqueous model solutions of histidine and N-acetyl-l-aspartate (NAA) enabled the assignment of an additional signal component at δ = 8 ppm of Cs in vivo to the amide group at the peptide bond. The visibility of this proton could result from hydrogen bonding which would agree with the anticipated stronger motional restriction of Cs. Referring to the observation that all dipolar-coupled multiplets resolved in localized in vivo ¹H NMR spectra of human m. gastrocnemius collapse simultaneously when the fibre structure is tilted towards the magic angle (θ ≈ 55°), a common model for molecular confinement in muscle tissue is proposed on the basis of an interaction of the studied metabolites with myocellular membrane phospholipids.     

Reinvestigation of the (4, 20) band of the O2 second negative system

High resolution absorption spectra of the (4, 20) band in the second negative system (A²Π_u–X²Π_g) of O₂⁺ cation were measured and analyzed in the range of 11 900–12 300 cm^–1 via optical heterodyne velocity modulation spectroscopy. Precise molecular constants of the levels involved were obtained by a nonlinear least-squares fitting procedure combining with our previous spectra of the (4, 19) and (6, 20) bands.     

In Vivo Detection of N-Coupled Protons in Rat Brain by ISIS Localization and Multiple-Quantum Editing

Three-dimensional image-selected in vivo spectroscopy (ISIS) was combined with phase-cycled ¹H–¹⁵N heteronuclear multiple-quantum coherence (HMQC) transfer NMR for localized selective observation of protons J-coupled to ¹⁵N in phantoms and in vivo. The ISIS–HMQC sequence, supplemented by jump–return water suppression, permitted localized selective observation of 2–5 μmol of [¹⁵N_indole]tryptophan, a precursor of the neurotransmitter serotonin, through the ¹⁵N-coupled proton in 20–40 min of acquisition in vitro at 4.7 T. In vivo, the amide proton of [5-¹⁵N]glutamine was selectively observed in the brain of spontaneously breathing ¹⁵NH₄⁺-infused rats, using a volume probe with homogeneous ¹H and ¹⁵N fields. Signal recovery after three-dimensional localization was 72–82% in phantoms and 59 ± 4% in vivo. The result demonstrates that localized selective observation of ¹⁵N-coupled protons, with complete cancellation of all other protons except water, can be achieved in spontaneously breathing animals by the ISIS–HMQC sequence. This sequence performs both volume selection and heteronuclear editing through an addition/subtraction scheme and predicts the highest intrinsic sensitivity for detection of ¹⁵N-coupled protons in the selected volume. The advantages and limitations of this method for in vivo application are compared to those of other localized editing techniques currently in use for non-exchanging protons.     

Distributions of transverse relaxation times for soft-solids measured in strongly inhomogeneous magnetic fields

The single-sided NMR-MOUSE sensor that operates in highly inhomogeneous magnetic fields is used to record a CPMG ¹H transverse relaxation decay by CPMG echo trains for a series of cross-linked natural rubber samples. Effective transverse relaxation rates 1/T_2,short and 1/T_2,long were determined by a bi-exponential fit. A linear dependence of transverse relaxation rates on cross-link density is observed for medium to large values of cross-link density. As an alternative to multi-exponential fits the possibility to analyze the dynamics of soft polymer network in terms of multi-exponential decays via the inverse Laplace transformation was studied. The transient regime and the effect of the T₁/T₂ ratio in inhomogeneous static and radiofrequency magnetic fields on the CPMG decays were studied numerically using a dedicated C++ program to simulate the temporal and spatial dependence of the CPMG response. A correction factor T₂/T_2,eff is derived as a function of the T₁/T₂ ratio from numerical simulations and compared with earlier results from two different well logging devices. High-resolution T₁–T₂ correlations maps are obtained by two-dimensional Laplace inversion of CPMG detected saturation recovery curves. The T₁–T₂ experimental correlations maps were corrected for the T₁/T₂ effect using the derived T₂/T_2,eff correction factor.     

Optimization of Three-Dimensional TROSY-Type HCCH NMR Correlation of Aromatic H–C Groups in Proteins

Improved methods for three-dimensional TROSY-Type HCCH correlation involving protons of negligible CSA are presented. The TROSY approach differs from the conventional approach of heteronuclear decoupling in evolution and detection periods by not mixing fast and slowly relaxing coherences and usually suppressing the former. Pervushin et al. (J. Am. Chem. Soc. 120, 6394–6400 (1998)) have proposed a 3D TROSY-type HCCH experiment where the TROSY approach is applied only in one of the ¹³C dimensions. A new pulse sequence applying the TROSY approach in both indirect dimensions is advantageous when the TROSY effect of the carbons is large or when a relatively high resolution is required. For lower resolutions or moderate TROSY effects we show that it is possible to combine the best of both worlds, namely to suppress heteronuclear couplings without mixing fast and slowly relaxing coherences while at the same time superimpose the two components and thus have both contribute to the detected signal. That is possible using the novel technique of Spin-State-Selective Time-Proportional Phase Incrementation (S³ TPPI). The new 3D S³ TPPI TROSY HCCH method is demonstrated on a ¹³C,¹⁵N-labeled protein sample, RAP 18–112 (N-terminal domain of α₂-macroglobulin receptor associated protein), at 750 MHz and average sensitivity enhancements of 10% are obtained for the cross peaks in comparison to methods based on conventional decoupling on one of the carbons or on TROSY on both carbons.     

An ab initio calculation of O and Si NMR parameters for SiO2 polymorphs

Ab initio molecular orbital calculations (Hartree–Fock, HF and density functional theories, DFTs) have been carried out for SiO₂ polymorphs coesite, low cristobalite, and α-quartz, in order to investigate the reliability of this method for predicting ²⁹Si and ¹⁷O nuclear magnetic resonance (NMR) properties of silicates. Oxygen- and silicon-centered clusters consisting of one (1T) to three tetrahedral (3T) shells (one to four atomic shells), taken from real crystal structure, have been investigated. It is found that for reasonable predication of both the ²⁹Si and ¹⁷O chemical shifts (δ_i^Si and δ_i^O), the minimum cluster is one that gives the correct second neighbors to the nucleus of interest. Both the δ_i^Si and δ_i^O have reached convergence with respect to cluster size at the OH-terminated two tetrahedral (2T) shell (three atomic shells around Si and four atomic shells around O) model. At convergence, the calculated δ_i^Si values agree well (within ±1 ppm) with experimental data. The calculated ¹⁷O electric field gradient (EFG)-related parameters also agree with experimental data within experimental uncertainties. The calculation also reproduces small differences in δ_i^O for O sites with similar tetrahedral connectivities, but shows deviations up to about 10 ppm in relative difference for O sites with different tetrahedral connectivities. The poor performance for the latter is mainly due to the approximations of the HF method. Our study thus suggests that the ab initio calculation method is a reliable mean for predicting ²⁹Si and ¹⁷O NMR parameters for silicates. Such an approach should find application not only to well-ordered crystalline phases, but also to disordered materials, by combining with other techniques, such as the molecular dynamics simulation method.     

Sensitivity of nuclear-quadrupole double-resonance detection of half-integer spin nuclei

The sensitivity of the Slusher and Hahn’s nuclear quadrupole double resonance technique is calculated in general for an arbitrary nuclear spin S of the quadrupole nuclei and for an arbitrary asymmetry parameter η of the electric field gradient tensor. The nuclear spin S = 5/2 (¹⁷O, ²⁵Mg, …) is treated in details. The influence of the cross-relaxation rate between the quadrupole nuclei and the abundant spin system on the sensitivity of double resonance is discussed. The results of the theoretical analysis are applied in the analysis of the ¹H–¹⁷O nuclear quadrupole double resonance spectra in p-toluenesulfonamide and 2-nitrobenzoic acid. The 17O nuclear quadrupole resonance frequencies from a sulfonamide group are determined for the first time. The proton–oxygen cross-relaxation rates and the proton local frequency in zero external magnetic field are experimentally determined from the nuclear quadrupole double resonance spectra.     

Sensitive, quantitative carbon-13 NMR spectra by mechanical sample translation

Collecting a truly quantitative carbon-13 spectrum is a time-consuming chore. Very long relaxation delays, required between transients to allow the z-magnetization, M_z, of the spin with the longest T₁ to return to the equilibrium value, M₀, must precede each transient. These long delays also reduce sensitivity, as fewer transients per unit time can be acquired. In addition, sometimes T₁ is not known to within even a factor of two: a conservative guess for the relaxation delay then leads to very low sensitivity. We demonstrate a fresh method to bypass these problems and collect quantitative carbon-13 spectra by swapping the sample volume after each acquisition with a different portion where the magnetization is already equilibrated to M₀. Loading larger sample volumes of 10–20 mL into an unusually long (1520 mm) 5 mm OD. NMR tube and vertically sliding the tube between acquisitions accomplishes the swap. The relaxation delay can then be skipped altogether. The spectra are thus both quantitative, and far more sensitive. We demonstrate the moving tube technique on two small molecules (thymol and butylhydroxytoluene) and show good carbon-13 quantification. The gain in sensitivity can be as much as 10-fold for slowly-relaxing ¹³C resonances. These experiments show that quantitative, sensitive carbon-13 spectra are possible whenever sufficient sample volumes are available. The method is applicable to any slow-relaxing nuclear spin species, such as ²⁹Si, ¹⁵N and other low-γ nuclei.     

Evolution of chirality from γ soft Ru to triaxial Ru

Rotational bands in ^108,110,112Ru have been investigated by means of γ–γ–γ and γ–γ(θ) coincidences of prompt γ rays emitted in the spontaneous fission of ²⁵²Cf. The positive parity bands are described by different versions of IBA, where ¹⁰⁸Ru is best described as a γ-soft nucleus whereas ^110,112Ru are more like rigid triaxial rotors. New ΔI=1 negative parity doublet bands are found. In case of ^110,112Ru, these are interpreted as soft chiral vibrations. Many of the experimental findings can be explained by microscopic calculations that combine the TAC mean-field with random phase approximation but a simple geometrical explanation is not apparent.