Magic Angle Spinning Conditions Research Articles

Experimental aspects of 14N overtone RESPDOR solid-state NMR spectroscopy under MAS beyond 60 kHz

Nitrogen-14 (14N) overtone (OT) spectroscopy under fast magic angle spinning (MAS) conditions (>60 kHz) has emerged as a powerful technique for observing correlations and distances between 14N and 1H, owing to the absence of the first-order quadrupolar broadenings. In addition, 14NOT allows selective manipulation of 14N nuclei for each site. Despite extensive theoretical and experimental studies, the spin dynamics of 14NOT remains under debate. In this study, we conducted experimental investigations to assess the spin dynamics of 14NOT using the rotational-echo saturation-pulse double-resonance (RESPDOR) sequence, which monitors population transfer induced by a14NOT pulse. The 14NOT spin dynamics is well represented by a model of a two-energy-level system. Unlike spin-1/2, the maximum excitation efficiency of 14NOT coherences of powdered solids, denoted by p, depends on the radiofrequency field (rf-field) strength due to orientation dependence of effective nutation fields even when pulse lengths are optimized. It is also found that the p factor, contributing to the 14NOT spin dynamics, is nearly independent of the B0 field. Consequently, the filtering efficiency of RESPDOR experiments exhibits negligible dependence on B0 when the 14NOT pulse length is optimized. The study also identifies the optimal experimental conditions for 14NOT/1H RESPDOR correlation experiments.

Opportunities and Challenges in Applying Solid-State NMR Spectroscopy in Organic Mechanochemistry.

In recent years it is shown that mechanochemical strategies can be beneficial in directed conversions of organic compounds. Finding new reactions proved difficult, and due to the lack of mechanistic understanding of mechanochemical reaction events, respective efforts have mostly remained empirical. Spectroscopic techniques are crucial in shedding light on these questions. In this overview, the opportunities and challenges of solid-state nuclear magnetic resonance (NMR) spectroscopy in the field of organic mechanochemistry are discussed. After a brief discussion of the basics of high-resolution solid-state NMR under magic-angle spinning (MAS) conditions, seven opportunities for solid-state NMR in the field of organic mechanochemistry are presented, ranging from ex situ approaches to structurally elucidated reaction products obtained by milling to the potential and limitations of in situ solid-state NMR approaches. Particular strengths of solid-state NMR, for instance in differentiating polymorphs, in NMR-crystallographic structure-determination protocols, or in detecting weak noncovalent interactions in molecular-recognition events employing proton-detected solid-state NMR experiments at fast MAS frequencies, are discussed.

Three Decades of REDOR in Protein Science: A Solid-State NMR Technique for Distance Measurement and Spectral Editing.

Solid-state NMR (ss-NMR) is a powerful tool to investigate noncrystallizable, poorly soluble molecular systems, such as membrane proteins, amyloids, and cell walls, in environments that closely resemble their physical sites of action. Rotational-echo double resonance (REDOR) is an ss-NMR methodology, which by reintroducing heteronuclear dipolar coupling under magic angle spinning conditions provides intramolecular and intermolecular distance restraints at the atomic level. In addition, REDOR can be exploited as a selection tool to filter spectra based on dipolar couplings. Used extensively as a spectroscopic ruler between isolated spins in site-specifically labeled systems and more recently as a building block in multidimensional ss-NMR pulse sequences allowing the simultaneous measurement of multiple distances, REDOR yields atomic-scale information on the structure and interaction of proteins. By extending REDOR to the determination of 1H-X dipolar couplings in recent years, the limit of measurable distances has reached ~15-20 Å, making it an attractive method of choice for the study of complex biomolecular assemblies. Following a methodological introduction including the most recent implementations, examples are discussed to illustrate the versatility of REDOR in the study of biological systems.

Identification of phosphorus sites in amorphous LiPON thin film by observing internuclear proximities

Amorphous lithium phosphorus oxynitrides (LiPON), prepared by reactive magnetron sputtering, have become the electrolytes of choice for all-solid-state thin film microbatteries since its discovery in early 1990s. Nevertheless, there is still a lack of understanding of their atomic-level structure and its influence on ionic conductivity. Solid-state NMR spectroscopy represents a promising technique to determine the atomic-level structure of LiPON glasses but is challenging owing to its low sensitivity in the case of thin film materials. Recently, 31P solid-state NMR spectra of LiPON thin films were acquired under magic-angle spinning (MAS) conditions and assigned with the help of density functional theory (DFT) calculations of NMR parameters. However, the identification of the different P local environments in these materials is still a challenge owing to their amorphous structure and the lack of resolution of the 31P MAS NMR spectra. We show herein how the NMR observation of internuclear proximities helps to establish the nature of P sites in LiPON thin films. The 31P-14N proximities are probed by a transfer of population in double resonance (TRAPDOR) experiment, whereas 31P-31P proximities are observed using one-dimensional (1D) 31P double-quantum (DQ)-filtered and two-dimensional (2D) 31P homonuclear correlation spectra as well as dipolar dephasing experiments using DQ-DRENAR (DQ-based dipolar-recoupling effects nuclear alignment reduction) technique. The obtained NMR data further support the recently proposed assignment of 31P NMR signals of LiPON thin films. With the help of this assignment, the simulation of the quantitative 1D 31P NMR spectrum indicates that PO43- orthophosphate anions prevail in LiPON thin films and N atoms are mainly incorporated in [O3PNPO3]5− dimeric anions. PO3N4− isolated tetrahedra and [O3POPO3]4− anions are also present but in smaller amounts.

Deuteron off-resonance rotating frame relaxation for the characterization of slow motions in rotating and static solid-state proteins

We demonstrate the feasibility of deuterium solid-state NMR off-resonance rotating frame relaxation measurements for studies of slow motions in biomolecular solids. The pulse sequence, which includes adiabatic pulses for magnetization alignment, is illustrated for static and magic-angle spinning conditions away from rotary resonances. We apply the measurements for three systems with selective deuterium labels at methyl groups: a) a model compound, Fluorenylmethyloxycarbonyl methionine-D3 amino acid, for which the principles of the measurements and corresponding motional modeling based on rotameric interconversions are demonstrated; b) amyloid-β1-40 fibrils labeled at a single alanine methyl group located in the disordered N-terminal domain. This system has been extensively studied in prior work and here serves as a test of the method for complex biological systems. The essential features of the dynamics consist of large-scale rearrangements of the disordered N-terminal domain and the conformational exchange between the free and bound forms of the domain, the latter one due to transient interactions with the structured core of the fibrils. and c) a 15-residue helical peptide which belongs to the predicted α-helical domain near the N-terminus of apolipoprotein B. The peptide is solvated with triolein and incorporates a selectively labeled leucine methyl groups. The method permits model refinement, indicating rotameric interconversions with a distribution of rate constants.

In-cell 31P solid-state NMR measurements of the lipid dynamics and influence of exogeneous β-amyloid peptides on live neuroblastoma neuro-2a cells

Non-specific disruption of cellular membranes induced by aggregation of exogeneous β-amyloid (Aβ) peptides is considered a viable pathological mechanism in Alzheimer's disease (AD). The solid-state nuclear magnetic resonance (ssNMR) spectroscopy has been widely applied in model liposomes to provide important insights on the molecular interactions between membranes and Aβ aggregates. Yet, the feasibility of in-cell ssNMR spectroscopy to probe Aβ-membrane interactions in native cellular environments has rarely been tested. Here we report the application of in-cell31P ssNMR spectroscopy on live mouse neuroblastoma Neuro-2a (N2a) cells under moderate magic angle spinning (MAS) conditions. Both cell viability and cytoplasmic membrane integrity are retained for up to six hours under 5 kHz MAS frequency at 277 K, which allow applications of direct-polarization 31P spectroscopy and 31P spin-spin (T2) relaxation measurements. The 31P T2 relaxation time constant of N2a cells is significantly increased compared with the model liposome prepared with comparable major phospholipid compositions. With the addition of 5 μM 40-residue Aβ (Aβ1–40) peptides, the 31P T2 relaxation is instantly accelerated. This work demonstrates the feasibility of using in-cell31P ssNMR to investigate the Aβ-membrane interactions in the biologically relevant cellular system.

CURD: a Single-Shot Strategy to Obtain Assignments and Distance Restraints for Proteins Using Solid-State MAS NMR Spectroscopy.

We present a strategy dubbed CURD (correlations using recycle delays) to acquire chemical-shift assignments and distance restraints for proteins in a single experimental block under slow-moderate magic-angle spinning conditions. This is done by concatenating the 3D-CCC and 3D-NNC experiments, both of which individually require long experimental times for sufficient resolution and sensitivity to be realized. Unlike previous approaches, the CURD strategy does not increase the amount of radio-frequency deposition on the sample and does not require lengthy procedures to optimize any of the pulse sequence elements. Instead, time savings is obtained by using the hitherto unused recycle delay of one of the experiments (2D-CC/3D-CCC) to establish inter-residue correlations for the second experiment (2D-NN/3D-NNC). Experiments are demonstrated on a model protein at the MAS frequency of 12.5 kHz and are shown to result in time savings of the order of days for most of the routine cases.

NMR Assignment of Methyl Groups in Immobilized Proteins Using Multiple-Bond 13C Homonuclear Transfers, Proton Detection, and Very Fast MAS.

In nuclear magnetic resonance spectroscopy of proteins, methyl protons play a particular role as extremely sensitive reporters on dynamics, allosteric effects, and protein–protein interactions, accessible even in high-molecular-weight systems approaching 1 MDa. The notorious issue of their chemical shift assignment is addressed here by a joint use of solid-state 1H-detected methods at very fast (nearly 100 kHz) magic-angle spinning, partial deuteration, and high-magnetic fields. The suitability of a series of RF schemes is evaluated for the efficient coherence transfer across entire 13C side chains of methyl-containing residues, which is key for establishing connection between methyl and backbone 1H resonances. The performance of ten methods for recoupling of either isotropic 13C–13C scalar or anisotropic dipolar interactions (five variants of TOBSY, FLOPSY, DIPSI, WALTZ, RFDR, and DREAM) is evaluated experimentally at two state-of-the-art magic-angle spinning (55 and 94.5 kHz) and static magnetic field conditions (18.8 and 23.5 T). Model isotopically labeled compounds (alanine and Met-Leu-Phe tripeptide) and ILV-methyl and amide-selectively protonated, and otherwise deuterated chicken α-spectrin SH3 protein are used as convenient reference systems. Spin dynamics simulations in SIMPSON are performed to determine optimal parameters of these RF schemes, up to recently experimentally attained spinning frequencies (200 kHz) and B 0 field strengths (28.2 T). The concept of linearization of 13C side chain by appropriate isotope labeling is revisited and showed to significantly increase sensitivity of methyl-to-backbone correlations. A resolution enhancement provided by 4D spectroscopy with non-uniform (sparse) sampling is demonstrated to remove ambiguities in simultaneous resonance assignment of methyl proton and carbon chemical shifts.

You cannot fight the pressure: Structural rearrangements of active pharmaceutical ingredients under magic angle spinning.

Although solid-state nuclear magnetic resonance (NMR) is a versatile analytical tool to study polymorphs and phase transitions of pharmaceutical molecules and products, this work summarizes examples of spontaneous and unexpected (and unwanted) structural rearrangements and phase transitions (amorphous-to-crystalline and crystalline-to-crystalline) under magic angle spinning (MAS) conditions, some of them clearly being due to the pressure experienced by the samples. It is widely known that such changes can often be detected by X-ray powder diffraction (XRPD); here, the capability of solid-state NMR experiments with a special focus on 1 H-13 C frequency-switched Lee-Goldburg heteronuclear correlation (FSLG HETCOR)/MAS NMR experiments to detect even subtle changes on a molecular level not observable by conventional 1D NMR experiments or XRPD is presented. Furthermore, it is shown that a polymorphic impurity combined with MAS can induce a crystalline-to-crystalline phase transition. This showcases that solid-state NMR is not always noninvasive and such changes upon MAS should be considered in particular when compounds are studied over longer time spans.

Reducing the effects of weak homonuclear dipolar coupling with CPMG pulse sequences for static and spinning solids

The Carr-Purcell/Meiboom-Gill (CPMG) pulse sequence, initially introduced for measuring transverse relaxation time constants (T2), can provide significant signal enhancements for solid-state NMR (SSNMR) spectra. The proper implementation of CPMG for acquiring spectra influenced by chemical shift anisotropies (CSAs), first and/or second order quadrupolar interactions, or paramagnetic broadening has been well documented to date, as have the effects of heteronuclear dipolar coupling on CPMG echo trains and T2 lifetimes. Homonuclear dipolar coupling can also impact T2 lifetimes and CPMG echo trains; these effects have been thoroughly investigated for spectra of homonuclear dipolar coupled spin-1/2 nuclei typically acquired under static conditions that are predominantly influenced by dipolar broadening (e.g., 1H, 19F, etc.). In particular, it has been shown that short refocusing pulses with small flip angles can extend the effective T2 (T2eff, the observed T2 constant as impacted by experimental conditions) measured by CPMG sequences for strong homonuclear dipolar coupled spin-1/2 pairs under static conditions. To date, these effects have not been explored for (i) spin-1/2 nuclei that have significant CSAs and simultaneously feature weak homonuclear dipolar couplings, (ii) for quadrupolar nuclei that are also weakly homonuclear dipolar coupled, and (iii) for either of these cases under magic-angle spinning (MAS) conditions. Herein, we demonstrate that short refocusing pulses that cause small flip angles can reduce the attenuation of signal in CPMG echo trains resulting from dipolar dephasing caused by the weak homonuclear dipolar couplings. For both spin-1/2 and quadrupolar nuclei, this can lead to significant extensions in T2eff and signal enhancements of up to three times compared to conventional CPMG in favourable cases. These phenomena can occur under both static and magic-angle spinning (MAS) conditions, in the latter of which homonuclear couplings are reintroduced by rotational resonance (R2) recoupling. Experimental examples of 13C (I = 1/2), 2H (I = 1), 87Rb (I = 3/2), 23Na (I = 3/2), and 35Cl (I = 3/2) NMR under static and MAS conditions, as well as simulations of these phenomena, are shown and discussed.

Deuteron rotating frame relaxation for the detection of slow motions in rotating solids

We demonstrate experimental and computational approaches for measuring 2H rotating frame NMR relaxation for solid samples under magic angle spinning (MAS) conditions. The relaxation properties of the deuterium spin-1 system are dominated by the reorientation of the anisotropic quadrupolar tensors, with the effective quadrupolar coupling constant around 55 kHz for methyl groups. The technique is demonstrated using the model compound dimethyl-sulfone at MAS rates of 10 and 60 kHz as well as for an amyloid fibril sample comprising an amyloid-β (1–40) protein with a selective methyl group labeled in the disordered domain of the fibrils, at an MAS rate of 8 kHz. For both systems, the motional parameters fall well within the ranges determined by other techniques, thus validating its feasibility. Experimental and computational factors such as i) the probe’s radio frequency inhomogeneity profiles, ii) rotary resonances at conditions for which the spin-lock field strength matches the half- or full-integer of the MAS rate, iii) the choice of MAS rates and spin-lock field strengths, and iv) simulations that account for the interconversion of multiple coherences for the spin-1 system under MAS and deviations from the analytical Redfield treatment are thoroughly considered.

On the potential of Fourier-encoded saturation transfers for sensitizing solid-state magic-angle spinning NMR experiments.

Chemical exchange saturation transfer (CEST) is widely used for enhancing the solution nuclear magnetic resonance (NMR) signatures of magnetically dilute spin pools, in particular, species at low concentrations undergoing chemical exchanges with an abundant spin pool. CEST's main feature involves encoding and then detecting weak NMR signals of the magnetically dilute spin pools on a magnetically abundant spin pool of much easier detection, for instance, the protons of H2O. Inspired by this method, we propose and exemplify a methodology to enhance the sensitivity of magic-angle spinning (MAS) solid-state NMR spectra. Our proposal uses the abundant 1H reservoir arising in organic solids as the magnetically abundant spin pool and relies on proton spin diffusion in lieu of chemical exchange to mediate polarization transfer between a magnetically dilute spin pool and this magnetically abundant spin reporter. As an initial test of this idea, we target the spectroscopy of naturally abundant 13C and rely on a Fourier-encoded version of the CEST experiment for achieving broadbandness in coordination with both MAS and heteronuclear decoupling, features normally absent in CEST. Arbitrary evolutions of multiple 13C sites can, thus, be imprinted on the entire 1H reservoir, which is subsequently detected. Theoretical predictions suggest that orders-of-magnitude signal enhancements should be achievable in this manner, on the order of the ratio between the 13C and the 1H reservoirs' abundances. Experiments carried out under magic-angle spinning conditions evidenced 5-10× gains in signal amplitudes. Further opportunities and challenges arising in this Fourier-encoded saturation transfer MAS NMR approach are briefly discussed.

Double echo symmetry-based REDOR and RESPDOR pulse sequences for proton detected measurements of heteronuclear dipolar coupling constants

1H{X} symmetry-based rotational echo double resonance pulse sequences (S-REDOR) and symmetry-based rotational echo saturation pulse double resonance (S-RESPDOR) solid-state NMR experiments have found widespread application for 1H detected measurements of difference NMR spectra, dipolar coupling constants, and internuclear distances under conditions of fast magic angle spinning (MAS). In these experiments the supercycled R412 (SR412) symmetry-based recoupling pulse sequence is typically applied to the 1H spins to reintroduce heteronuclear dipolar couplings. However, the timing of SR412 and other symmetry-based pulse sequences must be precisely synchronized with the rotation of the sample, otherwise, the evolution of 1H CSA and other interactions will not be properly refocused. For this reason, significant distortions are often observed in experimental dipolar dephasing difference curves obtained with S-REDOR or S-RESPDOR pulse sequences. Here we introduce a family of double echo (DE) S-REDOR/S-RESPDOR pulse sequences that function in an analogous manner to the recently introduced t1-noise eliminated (TONE) family of dipolar heteronuclear multiple quantum coherence (D-HMQC) pulse sequences. Through numerical simulations and experiments the DE S-REDOR/S-RESPDOR sequences are shown to provide dephasing difference curves similar to those obtained with S-REDOR/S-RESPDOR. However, the DE sequences are more robust to the deviations of the MAS frequency from the ideal value that occurs during typical solid-state NMR experiments. The DE sequences are shown to provide more reliable 1H detected dipolar dephasing difference curves for nuclei such as 15N (with isotopic labelling), 183W and 35Cl. The double echo sequences are therefore recommended to be used in place of conventional S-REDOR/S-RESPDOR sequences for measurement of weak dipolar coupling constants and long-range distances.

Silicon-29 echo train coherence lifetimes and geminal [formula omitted]-couplings in network modified silicate glasses

The natural abundance 29Si echo-train coherence lifetimes in network-modified silicate glasses were examined under static and magic-angle spinning (MAS) conditions. The nuclear magnetic properties of modifier cations were found to play a major role in determining 29Si coherence lifetimes, leading to differences as large as three orders of magnitude. In compositions with abundant NMR active nuclei, such as alkali silicates, the 29Si coherence lifetimes are dominated by coherent dephasing due to residual heteronuclear dipolar couplings, whereas in compositions dilute in NMR active nuclei, such as alkaline earth silicates, the 29Si coherence lifetimes are dominated by incoherent dephasing due to paramagnetic impurities. Expressing the inverse of the coherence lifetime as a residual full width at half maximum (FWHM), we found that increasing rates of both MAS and a π-pulse train are effective in removing the residual 29Si heteronuclear broadenings, with a near-linear relationship between FWHM and MAS rotor period and π-pulse spacing. Based on these results, we conclude that accurate 29Si J coupling measurements will be the most challenging in lithium silicate glasses due to strong homonuclear dipolar couplings among 7Li nuclei, requiring MAS speeds up to 100 kHz, and be the least challenging in the alkaline earth silicate glasses. At a modest MAS speed of 14kHz, distributions of geminal J couplings across Si-O-Si linkages were measured in alkali and alkaline earth silicate glasses giving mean values of 4.2Hz and 5.1Hz in 0.4 CaO·0.6 SiO2 and 0.33 Ba2O·0.67 SiO2 glasses, respectively, and 5.2Hz and 5.3Hz in 0.33 Na2O·0.67 SiO2 and 0.33 K2O·0.67 SiO2 glasses, respectively. We also observe greater variance in the J distributions of alkaline earth silicate glasses consistent with greater structural disorder due to increased modifier cation potential, i.e., the charge-to-radius ratio, Z/r of the cation.

Two- and Three-Dimensional 13C-17O Heteronuclear Correlation NMR Spectroscopy for Studying Organic and Biological Solids.

We report two- and three-dimensional (2D and 3D) 13C-17O heteronuclear correlation solid-state NMR experiments under magic-angle spinning (MAS) conditions. These experiments utilize the D-RINEPT (Dipolar-mediated Refocused Insensitive Nuclei Enhanced by Polarization Transfer) scheme with symmetry-based SR412 recoupling blocks for coherence transfer between 13C and 17O nuclei. First, a 2D 17O → 13C correlation experiment was performed for the [1-13C,17O]-Gly/Gly·HCl cocrystal and [U-13C, 1-17O]-α/β-d-glucose samples. Second, a 2D 17O → 13C MQ-D-RINEPT correlation experiment where the indirect dimension incorporates the multiple-quantum MAS (MQMAS) scheme was tested for obtaining isotropic 17O resolution with [U-13C, 1-17O]-α/β-d-glucose. Third, a new 3D 17O → 13C → 13C correlation experiment was demonstrated where 17O → 13C and 13C → 13C correlations are achieved by D-RINEPT and DARR (Dipolar Assisted Rotational Resonance) sequences, respectively (thus termed as a 3D D-RINEPT/DARR OCC experiment). This new 3D 17O NMR experiment is implemented with the aim for site-resolved solid-state 17O NMR studies.

Deuteron Chemical Exchange Saturation Transfer for the Detection of Slow Motions in Rotating Solids.

We utilized the 2H Chemical Exchange Saturation Transfer (CEST) technique under magic angle spinning (MAS) conditions to demonstrate the feasibility of the method for studies of slow motions in the solid state. For the quadrupolar anisotropic interaction, the essence of CEST is to scan the saturation pattern over a range of offsets corresponding to the entire spectral region(s) for all conformational states involved, which translates into a range of −60–+ 60 kHz for methyl groups. Rotary resonances occur when the offsets are at half-and full-integer of the MAS rates. The choice of the optimal MAS rate is governed by the condition to reduce the number of rotary resonances in the CEST profile patterns and retain a sufficiently large quadrupolar interaction active under MAS to maintain sensitivity to motions. As examples, we applied this technique to a well-known model compound dimethyl-sulfone (DMS) as well as amyloid-β fibrils selectively deuterated at a single methyl group of A2 belonging to the disordered domain. It is demonstrated that the obtained exchange rate between the two rotameric states of DMS at elevated temperatures fell within known ranges and the fitted model parameters for the fibrils agree well with the previously obtained value using static 2H NMR techniques. Additionally, for the fibrils we have observed characteristic broadening of rotary resonances in the presence of conformational exchange, which provides implications for model selection and refinement. This work sets the stage for future potential extensions of the 2H CEST under MAS technique to multiple-labeled samples in small molecules and proteins.

Frequency-Swept Ultra-Wideline Magic-Angle Spinning NMR Spectroscopy.

Recent years have witnessed the development of solid-state NMR techniques that allow the direct investigation of extremely wide inhomogeneously broadened resonance lines. To date, this typically involves the application of frequency sweeps as offered by wideband uniform rate smooth truncation (WURST) pulses. While the effects of such advanced irradiation schemes on static samples are well understood, the interference between the varying carrier frequency and the time-dependent evolution of the spin system under magic-angle spinning (MAS) conditions is more complex. Herein, we introduce the well-known WURST-Carr-Purcell-Meiboom-Gill (WCPMG) pulse sequence for spinning samples. Using numerical spin-density matrix analysis, an ideal design based on fast frequency sweeps and high truncation of the incorporated WURST pulses is presented that enables uniform excitation/refocusing under MAS conditions with low-to-moderate radio-frequency power requirements. This permits the acquisition of ultra-wideline MAS NMR lines exceeding 500 kHz with chemical shift resolution in a single transmitter step.

Long-Chain Fatty Acids in Bones and Their Link to Submicroscopic Vascularization Network: NMR Assignment and Relaxation Studies under Magic Angle Spinning Conditions in Intramuscular Bones of Atlantic Herring Fish.

The long-lasting proton signals in bones are identified as long-chain fatty acids, including saturated, mono-, and di-unsaturated fatty acids, with direct nuclear magnetic resonance evidence. We used intramuscular bones from Atlantic Herring fish to avoid interference from lipid-rich marrows. The key is to recognize that these signals are from mobile phase materials and study them with J-coupled correlation spectroscopies under magic angle spinning conditions. We kept extensive 1H-spin-echo records that allowed us to examine the effect of magic angle spinning on the transverse relaxation time of water and lipids over time. While it is impossible to distinguish based on chemical shifts, the relaxation data suggest that the signals are more consistent with the interpretation of phospholipid membranes than triglycerides in lipid droplets. In particular, the simultaneous T2 changes in water and lipids suggest that the centrifugal impact of magic angle spinning alters the lipid's structure in very tight spaces. Additional evidence of phospholipid membranes came from the choline-γ resonance at 3.2 ppm in fresh samples, which disappears with magic angle spinning. Thus, the fatty acid signals are at least partially from membrane bilayer structures, and we propose that they are linked to the submicroscopic vascularization channels similar to the dense canaliculi network in mammalian bones. Our detection of phospholipids from bones depended critically on two factors: (1) the elimination of the overwhelming triglyceride signals from marrows and (2) the preservation of water that biomembranes require. The relaxation data reveal aspects of lipid fluidity that have not been elucidated by previous order parameter studies on model membranes. Relaxation times have long been considered difficult to interpret. A robust and renewed understanding may be beneficial.

Through-space 11 B-27 Al correlation: Influence of the recoupling channel.

Through-space heteronuclear correlation (D-HETCOR) experiments based on heteronuclear multiple-quantum correlation (D-HMQC) and refocused insensitive nuclei enhanced by polarization transfer (D-RINEPT) sequences have been proven to be useful approaches for the detection of the spatial proximity between half-integer quadrupolar nuclei in solids under magic-angle spinning (MAS) conditions. The corresponding pulse sequences employ coherence transfers mediated by heteronuclear dipolar interactions, which are reintroduced under MAS by radiofrequency irradiation of only one of the two correlated nuclei. We investigate herein using numerical simulations of spin dynamics and solid-state NMR experiments on magnesium aluminoborate glass how the choice of the channel to which the heteronuclear dipolar recoupling is applied affects the transfer efficiency of D-HMQC and D-RINEPT sequences between 11 B and 27 Al nuclei. Experimental results show that maximum transfer efficiency is achieved when the recoupling scheme is applied to the channel, for which the spin magnetization is parallel to the B0 axis in average.

Broadband adiabatic inversion experiments for the measurement of longitudinal relaxation time constants.

Accurate measurements of longitudinal relaxation time constants (T1) in solid-state nuclear magnetic resonance (SSNMR) experiments are important for the study of molecular-level structure and dynamics. Such measurements are often made under magic-angle spinning conditions; however, there are numerous instances where they must be made on stationary samples, which often give rise to broad powder patterns arising from large anisotropic NMR interactions. In this work, we explore the use of wideband uniform-rate smooth-truncation pulses for the measurement of T1 constants. Two experiments are introduced: (i) BRAIN-CPT1, a modification of the BRAIN-CP (BRoadband Adiabatic-INversion-Cross Polarization) sequence, for broadband CP-based T1 measurements and (ii) WCPMG-IR, a modification of the WURST-CPMG sequence, for direct-excitation (DE) inversion-recovery experiments. A series of T1 constants are measured for spin-1/2 and quadrupolar nuclei with broad powder patterns, such as 119Sn (I = 1/2), 35Cl (I = 3/2), 2H (I = 1), and 195Pt (I = 1/2). High signal-to-noise spectra with uniform patterns can be obtained due to signal enhancements from T2 eff-weighted echo trains, and in favorable cases, BRAIN-CPT1 allows for the rapid measurement of T1 in comparison to DE experiments. Protocols for spectral acquisition, processing, and analysis of relaxation data are discussed. In most cases, relaxation behavior can be modeled with either monoexponential or biexponential functions based upon measurements of integrated powder pattern intensity; however, it is also demonstrated that one must interpret such T1 values with caution, as demonstrated by measurements of T1 anisotropy in 119Sn, 2H, and 195Pt NMR spectra.