Trace Level Detection and Quantification of Crystalline Silica in an Amorphous Silica Matrix with Natural Abundance 29Si NMR.

A protocol for the detection of trace amounts of quartz in amorphous silica gels by NMR spectroscopy was developed and tested on commercially available samples. Using natural abundance 29Si MAS NMR spectroscopy with CPMG acquisition and standard addition of crystalline quartz, quantitative detection of quartz concentrations down to 0.1% wt. was achieved. CPMG permitted to suppress the amorphous silica-derived signal, benefitting from the extremely long T2 relaxation time of quartz in 29Si and hence dramatically increasing the sensitivity. Dedicated post-processing exploiting the known CPMG spikelet frequencies allowed to probe the near-absence of quartz in commercial, 100% silica samples, enabling assessment of conformity of unknown samples to EU legislation (REACH).

Sensitivity improvement in 5QMAS NMR experiments using FAM-N pulses.

The multiple-quantum magic-angle spinning (MQMAS) experiment is a popular choice for obtaining high-resolution solid-state NMR spectra of quadrupolar nuclei with half-integer spin quantum number. However, its inherently poor sensitivity limits its application in more challenging systems. In particular, the use of higher-order multiple-quantum coherences, which have the potential to provide higher resolution in the isotropic spectrum, results in a further decrease in sensitivity. Here we extend our recent work, which introduced an automated, high-throughput approach to generate amplitude-modulated composite pulses (termed FAM-N) to improve the efficiency of the conversion of three-quantum coherences, and explore the use of similar pulses in five-quantum MAS experiments. We consider three different approaches, and are able to demonstrate that all three provide good enhancements over single pulse conversion in all but the most extreme cases, and work well at a range of spinning rates. We show that FAM-N pulses are robust to variation in the quadrupolar coupling and rf nutation rate, demonstrating their applicability in multisite systems and systems where direct experimental optimisation of complex composite pulses is not feasible. This work will ease the implementation of higher-order MQMAS experiments and enable their application to materials and systems that were previously deemed too difficult to study.

Resolution enhancement in proton double quantum magic-angle spinning spectra by constant-time acquisition.

Although very fast MAS rate (>60 kHz) paves a way to obtain a sufficient resolution in the 1H double-quantum magic-angle spinning (DQMAS) experiments to probe 1H proximities, the 1H resolution still limits wider applications below its potential use. Here, the combination of the DQMAS experiment with the constant-time (CT) acquisition approach is demonstrated, giving an increased peak-separation power in the DQ dimension. The advantages and disadvantages in terms of sensitivity and resolution of the conventional and CT approaches are discussed.

Investigating FAM-N pulses for signal enhancement in MQMAS NMR of quadrupolar nuclei.

Although a popular choice for obtaining high-resolution solid-state NMR spectra of quadrupolar nuclei, the inherently low sensitivity of the multiple-quantum magic-angle spinning (MQMAS) experiment has limited its application for nuclei with low receptivity or when the available sample volume is limited. A number of methods have been introduced in the literature to attempt to address this problem. Recently, we have introduced an alternative, automated approach, based on numerical simulations, for generating amplitude-modulated pulses (termed FAM-N pulses) to enhance the efficiency of the triple- to single-quantum conversion step within MQMAS. This results in efficient pulses that can be used without experimental reoptimisation, ensuring that this method is particularly suitable for challenging nuclei and systems. In this work, we investigate the applicability of FAM-N pulses to a wider variety of systems, and their robustness under more challenging experimental conditions. These include experiments performed under fast MAS, nuclei with higher spin quantum numbers, samples with multiple distinct sites, low-γ nuclei and nuclei subject to large quadrupolar interactions.

Nuclear overhauser spectroscopy of chiral CHD methylene groups.

Nuclear magnetic resonance spectroscopy (NMR) can provide a great deal of information about structure and dynamics of biomolecules. The quality of an NMR structure strongly depends on the number of experimental observables and on their accurate conversion into geometric restraints. When distance restraints are derived from nuclear Overhauser effect spectroscopy (NOESY), stereo-specific assignments of prochiral atoms can contribute significantly to the accuracy of NMR structures of proteins and nucleic acids. Here we introduce a series of NOESY-based pulse sequences that can assist in the assignment of chiral CHD methylene protons in random fractionally deuterated proteins. Partial deuteration suppresses spin-diffusion between the two protons of CH2 groups that normally impedes the distinction of cross-relaxation networks for these two protons in NOESY spectra. Three and four-dimensional spectra allow one to distinguish cross-relaxation pathways involving either of the two methylene protons so that one can obtain stereospecific assignments. In addition, the analysis provides a large number of stereospecific distance restraints. Non-uniform sampling was used to ensure optimal signal resolution in 4D spectra and reduce ambiguities of the assignments. Automatic assignment procedures were modified for efficient and accurate stereospecific assignments during automated structure calculations based on 3D spectra. The protocol was applied to calcium-loaded calbindin D9k. A large number of stereospecific assignments lead to a significant improvement of the accuracy of the structure.

Efficient amplitude-modulated pulses for triple- to single-quantum coherence conversion in MQMAS NMR.

The conversion between multiple- and single-quantum coherences is integral to many nuclear magnetic resonance (NMR) experiments of quadrupolar nuclei. This conversion is relatively inefficient when effected by a single pulse, and many composite pulse schemes have been developed to improve this efficiency. To provide the maximum improvement, such schemes typically require time-consuming experimental optimization. Here, we demonstrate an approach for generating amplitude-modulated pulses to enhance the efficiency of the triple- to single-quantum conversion. The optimization is performed using the SIMPSON and MATLAB packages and results in efficient pulses that can be used without experimental reoptimisation. Most significant signal enhancements are obtained when good estimates of the inherent radio-frequency nutation rate and the magnitude of the quadrupolar coupling are used as input to the optimization, but the pulses appear robust to reasonable variations in either parameter, producing significant enhancements compared to a single-pulse conversion, and also comparable or improved efficiency over other commonly used approaches. In all cases, the ease of implementation of our method is advantageous, particularly for cases with low sensitivity, where the improvement is most needed (e.g., low gyromagnetic ratio or high quadrupolar coupling). Our approach offers the potential to routinely improve the sensitivity of high-resolution NMR spectra of nuclei and systems that would, perhaps, otherwise be deemed "too challenging".

Understanding hydrogen-bonding structures of molecular crystals via electron and NMR nanocrystallography.

Understanding hydrogen-bonding networks in nanocrystals and microcrystals that are too small for X-ray diffractometry is a challenge. Although electron diffraction (ED) or electron 3D crystallography are applicable to determining the structures of such nanocrystals owing to their strong scattering power, these techniques still lead to ambiguities in the hydrogen atom positions and misassignments of atoms with similar atomic numbers such as carbon, nitrogen, and oxygen. Here, we propose a technique combining ED, solid-state NMR (SSNMR), and first-principles quantum calculations to overcome these limitations. The rotational ED method is first used to determine the positions of the non-hydrogen atoms, and SSNMR is then applied to ascertain the hydrogen atom positions and assign the carbon, nitrogen, and oxygen atoms via the NMR signals for 1H, 13C, 14N, and 15N with the aid of quantum computations. This approach elucidates the hydrogen-bonding networks in L-histidine and cimetidine form B whose structure was previously unknown.

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.