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Fluorine is becoming increasingly prevalent in medicinal chemistry, both in drug molecules and in molecular probes. The presence of fluorine allows convenient monitoring of such molecules in complex environments by NMR spectroscopy. However, sensitivity is a persistent limitation of NMR, especially when molecules are present at low concentrations. Here, sensitivity issues with 1H NMR are mitigated by sharing 19F photochemically-induced dynamic nuclear polarisation with 1H nuclei. Unlike direct 1H enhancement, this method enhances 1H signals without significantly distorting multiplet intensities, and has the potential to enable the use of suitable molecules as low-concentration probes. [ABSTRACT FROM AUTHOR]

Dynamic nuclear polarisation (DNP) is a process that transfers electron spin polarisation to nuclei by applying resonant microwave radiation, and has been widely used to improve the sensitivity of nuclear magnetic resonance (NMR). Here we demonstrate new levels of performance for static cross-effect proton DNP using high peak power chirped inversion pulses at 94 GHz to create a strong polarisation gradient across the inhomogeneously broadened line of the mono-radical 4-amino TEMPO. Enhancements of up to 340 are achieved at an average power of a few hundred mW, with fast build-up times (3 s). Experiments are performed using a home-built wideband kW pulsed electron paramagnetic resonance (EPR) spectrometer operating at 94 GHz, integrated with an NMR detection system. Simultaneous DNP and EPR characterisation of other mono-radicals and biradicals, as a function of temperature, leads to additional insights into limiting relaxation mechanisms and give further motivation for the development of wideband pulsed amplifiers for DNP at higher frequencies. Dynamic nuclear polarization is a technique to enhance NMR signals, often achieved by continuous-wave irradiation of the samples. Here, the authors create large polarization gradients by using high-power chirped inversion pulses at 94 GHz at an average power of a few hundred mW with fast build-up times, reaching NMR signal enhancements of up to 340. [ABSTRACT FROM AUTHOR]

A general theory is presented describing the photo-excitation dynamics of transient, paramagnetic triplet states of aromatic molecules including their absorption properties in a molecular host crystal when irradiated by a pulsed laser beam. It is applied to the system of a pentacene doped naphthalene single crystal, where the photo-excitation of pentacene results in a highly non-equilibrium population of intermediate electronic triplet states. These paramagnetic states can be addressed to align the proton spins of the naphthalene host at moderate experimental conditions using dynamic nuclear polarisation (DNP). The photo-excitation profile and the resulting triplet state distribution, which is determined by the material and the laser parameters, leads to a corresponding proton polarisation profile. Measurements of the resulting polarisation inhomogeneity by spatially resolved neutron transmission confirm the theory. The theoretical model then allows the extraction of crucial parameters that enable quantitative investigations of the dynamics of spin systems. For applications of triplet DNP with pentacene-doped naphthalene crystals, polarisation maximum, homogeneity and buildup rate are key factors, and the theory allows the identification of optimal experimental parameters depending on the priorities placed on these factors. [ABSTRACT FROM AUTHOR]

Hyperpolarization of 13C-enriched biomolecules via dissolution dynamic nuclear polarization (DNP) has enabled real-time metabolic imaging of a variety of diseases with superb specificity and sensitivity. The source of the unprecedented liquid-state nuclear magnetic resonance spectroscopic or imaging signal enhancements of >10 000-fold is the microwave-driven DNP process that occurs at a relatively high magnetic field and cryogenic temperature. Herein, we have methodically investigated the relative efficiencies of 13C DNP of single or double 13C-labeled sodium acetate with or without 2H-enrichment of the methyl group and using a 4-oxo-TEMPO free radical as the polarizing agent at 3.35 T and 1.4 K. The main finding of this work is that not all 13C spins in acetate are polarized with equal DNP efficiency using this relatively wide electron spin resonance linewidth free radical. In fact, the carbonyl 13C spins have about twice the solid-state 13C polarization level of methyl 13C spins. Deuteration of the methyl group provides a DNP signal improvement of methyl 13C spins on a par with that of carbonyl 13C spins. On the other hand, both the double 13C-labeled [1,2-13C2] acetate and [1,2-13C2, 2H3] acetate have a relative solid-state 13C polarization at the level of [2-13C] acetate. Meanwhile, the solid-state 13C T1 relaxation times at 3.35 T and 1.4 K were essentially the same for all six isotopomers of 13C acetate. These results suggest that the intramolecular environment of 13C spins plays a prominent role in determining the 13C DNP efficiency, while the solid phase 13C T1 relaxation of these samples is dominated by the paramagnetic effect due to the relatively high concentration of free radicals. [ABSTRACT FROM AUTHOR]

In contrast to the rapid and revolutionary impact of solution-state Nuclear Magnetic Resonance (NMR) on modern chemistry, the field of solid-state NMR has matured more slowly. This reflects the major technical challenges of much reduced spectral resolution and sensitivity in solid-state as compared to solution-state spectra, as well as the relative complexity of the solid state. In this perspective, we outline the technique developments that have pushed resolution to intrinsic limits and the approaches, including ongoing major developments in the field of Dynamic Nuclear Polarisation, that have enhanced spectral sensitivity. The information on local structure and dynamics that can be obtained using these gains in sensitivity and resolution is illustrated with a diverse range of examples from large biomolecules to energy materials and pharmaceuticals and from both ordered and highly disordered materials. We discuss how parallel developments in quantum chemical calculation, particularly density functional theory, have enabled experimental data to be translated directly into information on local structure and dynamics, giving rise to the developing field of "NMR crystallography." [ABSTRACT FROM AUTHOR]

We present a pulsed dynamic nuclear polarization (DNP) study using a ramped-amplitude nuclear orientation via electron spin locking (RA-NOVEL) sequence that utilizes a fast arbitrary waveform generator (AWG) to modulate the microwave pulses together with samples doped with narrow-line radicals such as 1,3-bisdiphenylene-2-phenylallyl (BDPA), sulfonated-BDPA (SA-BDPA), and trityl-OX063. Similar to ramped-amplitude cross polarization in solid-state nuclear magnetic resonance, RA-NOVEL improves the DNP efficiency by a factor of up to 1.6 compared to constant-amplitude NOVEL (CA-NOVEL) but requires a longer mixing time. For example, at τmix = 8 μs, the DNP efficiency reaches a plateau at a ramp amplitude of ~20 MHz for both SA-BDPA and trityl-OX063, regardless of the ramp profile (linear vs. tangent). At shorter mixing times (τ mix = 0.8 μs), we found that the tangent ramp is superior to its linear counterpart and in both cases there exists an optimum ramp size and therefore ramp rate. Our results suggest that RA-NOVEL should be used instead of CA-NOVEL as long as the electronic spin lattice relaxation T1e is sufficiently long and/or the duty cycle of the microwave amplifier is not exceeded. To the best of our knowledge, this is the first example of a time domain DNP experiment that utilizes modulated microwave pulses. Our results also suggest that a precise modulation of the microwave pulses can play an important role in optimizing the efficiency of pulsed DNP experiments and an AWG is an elegant instrumental solution for this purpose. [ABSTRACT FROM AUTHOR]

In situ illumination of liquid-state nuclear magnetic resonance (NMR) samples makes it possible for a wide range of light-dependent chemical and biological phenomena to be studied by the powerful analytical technique. However, the position of an NMR sample deep within the bore of the spectrometer magnet renders such illumination challenging. Here, we demonstrate the working principles of a sample illumination device (NMRtorch) where a lighthead containing an LED array is positioned directly at the top of an NMRtorch tube which is inserted into the NMR spectrometer. The wall of the tube itself acts as a light guide, illuminating the sample from the outside. We explore how this new setup performs in a number of photo-NMR applications, including photoisomerisation and photo-chemically induced dynamic nuclear polarisation (photo-CIDNP), and demonstrate the potential for ultraviolet (UV) degradation studies with continuous online NMR assessment. This setup enables users of any typical liquid-state spectrometer to easily perform in situ photo-NMR experiments, using a wide range of wavelengths. In situ illumination of liquid-state NMR samples allows to characterise light-dependent chemical and biological phenomena, but, in practice, the position of an NMR sample deep within the bore of a spectrometer magnet renders such illumination challenging. Here, the authors demonstrate the working principles of a sample illumination device, with an LED array positioned directly at the top of special sample tube, which is inserted into the NMR spectrometer. [ABSTRACT FROM AUTHOR]

Polarisation transfer schemes and indirect detection are central to magnetic resonance. Using the trityl radical OX063 and a pulse electron paramagnetic resonance spectrometer operating in the Q-band (35 GHz, 1.2 T), we show here that it is possible to use pulsed dynamic nuclear polarisation (DNP) to transfer polarisation from electrons to protons and back. The latter is achieved by first saturating the electrons and then simply using a reverse DNP step. A variable mixing time between DNP and reverse DNP allows us to investigate the decay of polarisation on protons in the vicinity of the electrons. We qualitatively investigate the influence of solvent deuteration, temperature, and electron concentration. We expect reverse DNP to be useful in the investigation of nuclear spin diffusion and envisage its use in electron–nuclear double-resonance (ENDOR) experiments. [ABSTRACT FROM AUTHOR]

Dissolution dynamic nuclear polarisation (dDNP) of 13C‐labelled pyruvate in magnetic resonance spectroscopy/imaging (MRS/MRSI) has the potential for monitoring tumour progression and treatment response. Pyruvate delivery, its metabolism to lactate and efflux were investigated in rat P22 sarcomas following simultaneous intravenous administration of hyperpolarised 13C‐labelled pyruvate (13C1‐pyruvate) and urea (13C‐urea), a nonmetabolised marker. A general mathematical model of pyruvate‐lactate exchange, incorporating an arterial input function (AIF), enabled the losses of pyruvate and lactate from tumour to be estimated, in addition to the clearance rate of pyruvate signal from blood into tumour, Kip, and the forward and reverse fractional rate constants for pyruvate‐lactate signal exchange, kpl and klp. An analogous model was developed for urea, enabling estimation of urea tumour losses and the blood clearance parameter, Kiu. A spectral fitting procedure to blood time‐course data proved superior to assuming a gamma‐variate form for the AIFs. Mean arterial blood pressure marginally correlated with clearance rates. Kiu equalled Kip, indicating equivalent permeability of the tumour vasculature to urea and pyruvate. Fractional loss rate constants due to effluxes of pyruvate, lactate and urea from tumour tissue into blood (kpo, klo and kuo, respectively) indicated that T1s and the average flip angle, θ, obtained from arterial blood were poor surrogates for these parameters in tumour tissue. A precursor‐product model, using the tumour pyruvate signal time‐course as the input for the corresponding lactate signal time‐course, was modified to account for the observed delay between them. The corresponding fractional rate constant, kavail, most likely reflected heterogeneous tumour microcirculation. Loss parameters, estimated from this model with different TRs, provided a lower limit on the estimates of tumour T1 for lactate and urea. The results do not support use of hyperpolarised urea for providing information on the tumour microcirculation over and above what can be obtained from pyruvate alone. The results also highlight the need for rigorous processes controlling signal quantitation, if absolute estimations of biological parameters are required. [ABSTRACT FROM AUTHOR]

We report the generation and observation of long-lived spin states in deuterated methyl groups by dissolution DNP. These states are based on population imbalances between manifolds of spin states corresponding to irreducible representations of the C3v point group and feature strongly dampened quadrupolar relaxation. Their lifetime depends on the activation energies of methyl group rotation. With dissolution DNP, we can reduce the deuterium relaxation rate by a factor up to 20, thereby extending the experimentally available time window. The intrinsic limitation of NMR spectroscopy of quadrupolar spins by short relaxation times can thus be alleviated. [ABSTRACT FROM AUTHOR]

Dynamic nuclear polarization (DNP) is a technique that uses a microwave-driven transfer of high spin alignment from electrons to nuclear spins. This is most effective at low temperature and high magnetic field, and with the invention of the dissolution method, the amplified nuclear magnetic resonance (NMR) signals in the frozen state in DNP can be harnessed in the liquid-state at physiologically acceptable temperature for in vitro and in vivo metabolic studies. A current optimization practice in dissolution DNP is to dope the sample with trace amounts of lanthanides such as Gd3+ or Ho3+, which further improves the polarization. While Gd3+ and Ho3+ have been optimized for use in dissolution DNP, other lanthanides have not been exhaustively studied for use in 13C DNP applications. In this work, two additional lanthanides with relatively high magnetic moments, Dy3+ and Tb3+, were extensively optimized and tested as doping additives for 13C DNP at 3.35 T and 1.2 K. We have found that both of these lanthanides are also beneficial additives, to a varying degree, for 13C DNP. The optimal concentrations of Dy3+ (1.5 mM) and Tb3+ (0.25 mM) for 13C DNP were found to be less than that of Gd3+ (2 mM). W-band electron paramagnetic resonance shows that these enhancements due to Dy3+ and Tb3+ doping are accompanied by shortening of electron T1 of trityl OX063 free radical. Furthermore, when dissolutionwas employed, Tb3+-doped samples were found to have similar liquid-state 13C NMRsignal enhancements compared to samples doped with Gd3+, and both Tb3+ and Dy3+ had a negligible liquid-state nuclear T1 shortening effect which contrasts with the significant reduction in T1 when using Gd3+. Our results showthat Dy3+ doping and Tb3+ doping have a beneficial impact on 13C DNP both in the solid and liquid states, and that Tb3+ in particular could be used as a potential alternative to Gd3+ in 13C dissolution DNP experiments. [ABSTRACT FROM AUTHOR]

We present results of a pulsed dynamic nuclear polarization (DNP) study at 0.35 T (9.7 GHz/ 14.7 MHz for electron/¹H Larmor frequency) using a lab frame-rotating frame cross polarization experiment that employs electron spin locking fields that match the ¹H nuclear Larmor frequency, the so called NOVEL (nuclear orientation via electron spin locking) condition. We apply the method to a series of DNP samples including a single crystal of diphenyl nitroxide (DPNO) doped benzophenone (BzP), 1,3-bisdiphenylene-2-phenylallyl (BDPA) doped polystyrene (PS), and sulfonated-BDPA (SA-BDPA) doped glycerol/water glassy matrices. The optimal Hartman-Hahn matching condition is achieved when the nutation frequency of the electron matches the Larmor frequency of the proton, ω1S = ω0I, together with possible higher order matching conditions at lower efficiencies. The magnetization transfer from electron to protons occurs on the time scale of ~100 ns, consistent with the electron-proton couplings on the order of 1-10 MHz in these samples. In a fully protonated single crystal DPNO/BzP, at 270 K, we obtained a maximum signal enhancement of e = 165 and the corresponding gain in sensitivity of e(T1/TB)1/2 = 230 due to the reduction in the buildup time under DNP. In a sample of partially deuterated PS doped with BDPA, we obtained an enhancement of 323 which is a factor of ~3.2 higher compared to the protonated version of the same sample and accounts for 49% of the theoretical limit. For the SA-BDPA doped glycerol/water glassy matrix at 80 K, the sample condition used in most applications of DNP in nuclear magnetic resonance, we also observed a significant enhancement. Our findings demonstrate that pulsed DNP via the NOVEL sequence is highly efficient and can potentially surpass continuous wave DNP mechanisms such as the solid effect and cross effect which scale unfavorably with increasing magnetic field. Furthermore, pulsed DNP is also a promising avenue for DNP at high temperature. [ABSTRACT FROM AUTHOR]

We develop a theoretical description of Dynamic Nuclear Polarization (DNP) in solids under Magic Angle Spinning (MAS) to describe the magnetic field dependence of the DNP effect. The treatment is based on an efficient scheme for numerical solution of the Liouville-von Neumann equation, which explicitly takes into account the variation of magnetic interactions during the sample spinning. The dependence of the cross-effect MAS-DNP on various parameters, such as the hyperfine interaction, electron-electron dipolar interaction, microwave field strength, and electron spin relaxation rates, is analyzed. Electron spin relaxation rates are determined by electron paramagnetic resonance measurements, and calculations are compared to experimental data. Our results suggest that the observed nuclear magnetic resonance signal enhancements provided by MAS-DNP can be explained by discriminating between "bulk" and "core" nuclei and by taking into account the slow DNP build-up rate for the bulk nuclei. [ABSTRACT FROM AUTHOR]

Signal amplification by reversible exchange (SABRE) offers a cost‐effective route to boost nuclear magnetic resonance (NMR) signal by several orders of magnitude by employing readily available para‐hydrogen as a source of hyperpolarisation. Although 1H spins have been the natural choice of SABRE hyperpolarisation since its inception due to its simplicity and accessibility, limited spin lifetimes of 1H makes it harder to employ them in a range of time‐dependent NMR experiments. Heteronuclear spins, for example, 13C and 15N, in general have much longer T1 lifetimes and thereby are found to be more suitable for hyperpolarised biological applications as demonstrated previously by para‐hydrogen induced polarisation (PHIP) and dynamic nuclear polarisation (DNP). In this study we demonstrate a simple procedure to enhance 15N signal of an antibiotic drug ornidazole by up to 71,000‐folds with net 15N polarisation reaching ~23%. Further, the effect of co‐ligand strategy is studied in conjunction with the optimum field transfer protocols and consequently achieving 15N hyperpolarised spin lifetime of >3 min at low field. Finally, we present a convenient route to harness the hyperpolarised solution in aqueous medium free from catalyst contamination leading to a strong 15N signal detection for an extended duration of time. [ABSTRACT FROM AUTHOR]

Signal-enhancement techniques for NMR spectroscopy are important to amplify the weak resonances provided by nuclear spins. Recently, 'hyperpolarization' techniques have been intensively investigated. These provide nuclear spin states far from equilibrium yielding strong signal boosts up to four orders of magnitude. Here we propose a method for real-time NMR of 'hyperpolarized' proteins at residue resolution. The approach is based on dissolution dynamic nuclear polarization (d-DNP), which enables the use of hyperpolarized buffers that selectively boost NMR signals of solvent-exposed protein residues. The resulting spectral sparseness and signal enhancements enable recording of residue-resolved spectra at a 2 Hz sampling rate. Thus, we monitor the hyperpolarization level of different protein residues simultaneously under near-physiological conditions. We aim to address two points: 1) NMR experiments are often performed under conditions that increase sensitivity but are physiologically irrelevant; 2) long signal accumulation impedes fast real-time monitoring. Both limitations are of fundamental relevance to ascertain pharmacological relevance and study protein kinetics. Dissolution dynamic nuclear polarisation allows for dramatic signal enhancement in protein NMR spectroscopy, but loss of polarisation limits temporal and structural resolution. Here polarisation of the solvent and subsequent transfer to the target molecule enables selective detection of hyperpolarised residues. [ABSTRACT FROM AUTHOR]

We show that TOCSY and multiple-quantum (MQ) 2D NMR spectra can be obtained for mixtures of substrates hyperpolarised by dissolution dynamic nuclear polarisation (D-DNP). This is achieved by combining optimised transfer settings for D-DNP, with ultrafast 2D NMR experiments based on spatiotemporal encoding. TOCSY and MQ experiments are particularly well suited for mixture analysis, and this approach opens the way to significant sensitivity gains for analytical applications of NMR, such as authentication and metabolomics. [ABSTRACT FROM AUTHOR]

In dissolution-dynamic nuclear polarization, a hyperpolarized solid is dissolved with a jet of hot solvent. The solution is then transferred to a secondary magnet, where spectra can be recorded with improved sensitivity. In bullet-dynamic nuclear polarization this order is reversed. Pressurized gas is used to rapidly transfer the hyperpolarized solid to the secondary magnet, and the hyperpolarized solid is dissolved only upon arrival. A potential advantage of this approach is that it may avoid excessive dilution and the associated signal loss, in particular for small sample quantities. Previously, we have shown that liquid-state NMR spectra with polarization levels of up to 30% may be recorded within less than 1 s after the departure of the hyperpolarized solid from the polarizing magnet. The resolution of the recorded spectra however was limited. The system consumed significant amounts of liquid helium, and substantial manual work was required in between experiments to prepare for the next shot. Here, we present a new bullet-DNP (dynamic nuclear polarization) system that addresses these limitations. [ABSTRACT FROM AUTHOR]

We report magic angle spinning, dynamic nuclear polarization (DNP) experiments at magnetic fields of 9.4 T, 14.1 T, and 18.8 T using the narrow line polarizing agents 1,3-bisdiphenylene-2-phenylallyl (BDPA) dispersed in polystyrene, and sulfonated-BDPA (SA-BDPA) and trityl OX063 in glassy glycerol/water matrices. The ¹H DNP enhancement field profiles of the BDPA radicals exhibit a significant DNP Overhauser effect (OE) as well as a solid effect (SE) despite the fact that these samples are insulating solids. In contrast, trityl exhibits only a SE enhancement. Data suggest that the appearance of the OE is due to rather strong electron-nuclear hyperfine couplings present in BDPA and SA-BDPA, which are absent in trityl and perdeuterated BDPA (d21 -BDPA). In addition, and in contrast to other DNP mechanisms such as the solid effect or cross effect, the experimental data suggest that the OE in non-conducting solids scales favorably with magnetic field, increasing in magnitude in going from 5 T, to 9.4 T, to 14.1 T, and to 18.8 T. Simulations using a model two spin system consisting of an electron hyperfine coupled to a ¹H reproduce the essential features of the field profiles and indicate that the OE in these samples originates from the zero and double quantum cross relaxation induced by fluctuating hyperfine interactions between the intramolecular delocalized unpaired electrons and their neighboring nuclei, and that the size of these hyperfine couplings is crucial to the magnitude of the enhancements. Microwave power dependent studies show that the OE saturates at considerably lower power levels than the solid effect in the same samples. Our results provide new insights into the mechanism of the Overhauser effect, and also provide a new approach to perform DNP experiments in chemical, biophysical, and physical systems at high magnetic fields. [ABSTRACT FROM AUTHOR]

Ionic liquids, and their solid-state equivalents organic ionic plastic crystals, show many useful and tailorable properties that make them interesting for a wide range of applications including as electrolytes for energy storage devices. Nuclear magnetic resonance spectroscopy and related techniques offer a powerful and versatile toolkit for the characterisation of structure, interactions and dynamics in these materials. This article summarises both commonly used methods and some recent advances in this area, including solution- and solid-state methods, dynamic nuclear polarisation, imaging, diffusion and relaxation measurements, and example applications of some less commonly studied nuclei. [ABSTRACT FROM AUTHOR]

Objectives: To enhance detection of the products of hyperpolarized [2-13C]dihydroxyacetone metabolism for assessment of three metabolic pathways in the liver in vivo. Hyperpolarized [2-13C]DHAc emerged as a promising substrate to follow gluconeogenesis, glycolysis and the glycerol pathways. However, the use of [2-13C]DHAc in vivo has not taken off because (i) the chemical shift range of [2-13C]DHAc and its metabolic products span over 144 ppm, and (ii) 1H decoupling is required to increase spectral resolution and sensitivity. While these issues are trivial for high-field vertical-bore NMR spectrometers, horizontal-bore small-animal MR scanners are seldom equipped for such experiments. Methods: Real-time hepatic metabolism of three fed mice was probed by 1H-decoupled 13C-MR following injection of hyperpolarized [2-13C]DHAc. The spectra of [2-13C]DHAc and its metabolic products were acquired in a 7 T small-animal MR scanner using three purpose-designed spectral-spatial radiofrequency pulses that excited a spatial bandwidth of 8 mm with varying spectral bandwidths and central frequencies (chemical shifts). Results: The metabolic products detected in vivo include glycerol 3-phosphate, glycerol, phosphoenolpyruvate, lactate, alanine, glyceraldehyde 3-phosphate and glucose 6-phosphate. The metabolite-to-substrate ratios were comparable to those reported previously in perfused liver. Discussion: Three metabolic pathways can be probed simultaneously in the mouse liver in vivo, in real time, using hyperpolarized DHAc. [ABSTRACT FROM AUTHOR]

While lithium metal represents the ultimate high-energy-density battery anode material, its use is limited by dendrite formation and associated safety risks, motivating studies of the solid–electrolyte interphase layer that forms on the lithium, which is key in controlling lithium metal deposition. Dynamic nuclear polarisation enhanced NMR can provide important structural information; however, typical exogenous dynamic nuclear polarisation experiments, in which organic radicals are added to the sample, require cryogenic sample cooling and are not selective for the interface between the metal and the solid–electrolyte interphase. Here we instead exploit the conduction electrons of lithium metal to achieve an order of magnitude hyperpolarisation at room temperature. We enhance the 7Li, 1H and 19F NMR spectra of solid–electrolyte interphase species selectively, revealing their chemical nature and spatial distribution. These experiments pave the way for more ambitious room temperature in situ dynamic nuclear polarisation studies of batteries and the selective enhancement of metal–solid interfaces in a wider range of systems. Understanding the solid–electrolyte interphase (SEI) is key to developing safe dendrite-free lithium batteries. Here, by exploiting the electrons in lithium metal to selectively hyperpolarise the NMR signals, the authors reveal the chemistry and spatial distribution of species at the metal–SEI interface. [ABSTRACT FROM AUTHOR]

Hyperpolarised [1‐13C]pyruvate MRI has shown promise in monitoring therapeutic efficacy in a number of cancers including glioma. In this study, we assessed the pyruvate response to the lentiviral suicide gene therapy of herpes simplex virus‐1 thymidine kinase with the prodrug ganciclovir (HSV‐TK/GCV) in C6 rat glioma and compared it with traditional MR therapy markers. Female Wistar rats were inoculated with 106 C6 glioma cells. Treated animals received intratumoural lentiviral HSV‐TK gene transfers on days 7 and 8 followed by 2‐week GCV therapy starting on day 10. Animals were repeatedly imaged during therapy using volumetric MRI, diffusion and relaxation mapping, as well as metabolic [1‐13C]pyruvate MRS imaging. Survival (measured as time before animals reached a humane endpoint and were euthanised) was assessed up to day 30 posttherapy. HSV‐TK/GCV gene therapy lengthened the median survival time from 12 to 25 days. This was accompanied by an apparent tumour growth arrest, but no changes in diffusion or relaxation parameters in treated animals. The metabolic response was more evident in the case‐by‐case analysis than in the group‐level analysis. Treated animals also showed a 37 ± 15% decrease (P < 0.05, n = 5) in lactate‐to‐pyruvate ratio between therapy weeks, whereas a 44 ± 18% increase (P < 0.05, n = 6) was observed in control animals. Hyperpolarised [1‐13C]pyruvate MRI can offer complementary metabolic information to traditional MR methods to give a more comprehensive picture of the slowly developing gene therapy response. This may benefit the detection of the successful therapy response in patients. [ABSTRACT FROM AUTHOR]

Electron spectral diffusion (eSD) plays an important role in solid-state, static dynamic nuclear polarization (DNP) with polarizers that have inhomogeneously broadened EPR spectra, such as nitroxide radicals. It affects the electron spin polarization gradient within the EPR spectrum during microwave irradiation and thereby determines the effectiveness of the DNP process via the so-called indirect cross-effect (iCE) mechanism. The electron depolarization profile can be measured by electron-electron double resonance (ELDOR) experiments, and a theoretical framework for deriving eSD parameters from ELDOR spectra and employing them to calculate DNP profiles has been developed. The inclusion of electron depolarization arising from the 14N solid effect (SE) has not yet been taken into account in this theoretical framework and is the subject of the present work. The 14N SE depolarization was studied using W-band ELDOR of a 0.5mM TEMPOL solution, where eSD is negligible, taking into account the hyperfine interaction of both 14N and 1H nuclei, the long microwave irradiation applied under DNP conditions, and electron and nuclear relaxation. The results of this analysis were then used in simulations of ELDOR spectra of 10 and 20mM TEMPOL solutions, where eSD is significant using the eSD model and the SE contributions were added ad hoc employing the 1H and 14N frequencies and their combinations, as found from the analysis of the 0.5mM sample. This approach worked well for the 20mM solution, where a good fit for all ELDOR spectra recorded along the EPR spectrum was obtained and the inclusion of the 14N SE mechanism improved the agreement with the experimental spectra. For the 10mM solution, simulations of the ELDOR spectra recorded along the gz position gave a lower-quality fit than for spectra recorded in the center of the EPR spectrum. This indicates that the simple approach we used to describe the 14N SE is limited when its contribution is relatively high as the anisotropy of its magnetic interactions was not considered explicitly. [ABSTRACT FROM AUTHOR]

Purpose: The objective of the present work was to develop and implement an efficient approach to hyperpolarize [1‐13C]acetate and apply it to in vivo cardiac spectroscopy and imaging. Methods: Rapid hydrogen peroxide induced decarboxylation was used to convert hyperpolarized [2‐13C]pyruvate into highly polarized [1‐13C]acetate employing an additional step following rapid dissolution of [2‐13C]pyruvate in a home‐built multi‐sample dissolution dynamic nuclear polarization system. Phantom dissolution experiments were conducted to determine optimal parameters of the decarboxylation reaction, retaining polarization and T1 of [1‐13C]acetate. In vivo feasibility of detecting [1‐13C]acetate metabolism is demonstrated using slice‐selective spectroscopy and multi‐echo imaging of [1‐13C]acetate and [1‐13C]acetylcarnitine in the healthy rat heart. Results: The first in vivo signal was observed ~23 s after dissolution. At the corresponding time point in the phantom experiments, 97.9 ± 0.4% of [2‐13C]pyruvate were converted into [1‐13C]acetate by the decarboxylation reaction. T1 and polarization of [1‐13C]acetate was determined to be 29.7 ± 1.9% and a 47.7 ± 0.5 s. Polarization levels of [2‐13C]pyruvate and [1‐13C]acetate were not significantly different after transfer to the scanner. In vivo, [1‐13C]acetate and [1‐13C]acetylcarnitine could be detected using spectroscopy and imaging. Conclusion: Decarboxylation of hyperpolarized [2‐13C]pyruvate enables the efficient production of highly polarized [1‐13C]acetate that is applicable to study short‐chain fatty acid metabolism in the in vivo heart. [ABSTRACT FROM AUTHOR]

• Circularly polarised RF selectively induces either positive or negative enhancements. • O-DNP with circularly polarised RF renders an enhancement factor of around 150,000 at 1.2 μT. • Images with resolution of close to 1 mm were obtained using O-DNP in microtesla fields. Magnetic resonance imaging in ultra-low fields is often limited by mediocre signal-to-noise ratio hindering a higher resolution. Overhauser dynamic nuclear polarisation (O-DNP) using nitroxide radicals has been an efficient solution for enhancing the thermal nuclear polarisation. However, the concurrence of positive and negative polarisation enhancements arises in ultra-low fields resulting in a significantly reduced net enhancement, making O-DNP far less attractive. Here, we address this issue by applying circularly polarised RF. O-DNP with circularly polarised RF renders a considerably improved enhancement factor of around 150,000 at 1.2 μ T. A birdcage coil was adopted into an ultra-low field MRI system to generate the circularly polarised RF field homogeneously over a large volume. We acquired an MR image of a nitroxide radical solution with an average in-plane resolution of 1 mm. De-noising through compressive sensing further improved the image quality. [ABSTRACT FROM AUTHOR]

Overhauser dynamic nuclear polarisation (DNP) represents a potentially outstanding tool to increase the sensitivity of solution and solid state NMR experiments, as well as of magnetic resonance imaging. DNP signal enhancements are strongly linked to the spin relaxation properties of the system under investigation, which must contain a paramagnetic molecule used as DNP polariser. In turn, nuclear spin relaxation can be monitored through NMR relaxometry, which reports on the field dependence of the nuclear relaxation rates, opening a route to understand the physical processes at the origin of the Overhauser DNP in solution. The contributions of dipole-dipole and Fermi-contact interactions to paramagnetic relaxation are here described and shown to be responsible to both the relaxometry profiles and the DNP enhancements, so that the experimental access to the former can allow for predictions of the latter. [ABSTRACT FROM AUTHOR]

Abstract Selenate-loaded selenium water remediation materials based on polymer fibres have been investigated by dynamic nuclear polarization (DNP) enhanced solid-state NMR. For carbon-13 a significant reduction in experiment time is obtained with DNP even when compared with conventional carbon-13 NMR spectra recorded using larger samples. For the selenium remediation materials studied here this reduction allows efficient acquisition of {1H}-77Se heteronuclear correlation spectra which give information about the nature of the binding of the remediated selenate ions with the grafted side chains which provide the required ion exchange functionality. Graphical abstract Image 1 Highlights • DNP allows efficient acquisition of {1H}-77Se heteronuclear correlation NMR spectra. • Even at relatively low Se concentrations in polymer water remediation materials. • Suggest a supramolecular interaction with the functionalizing polymer side chain. [ABSTRACT FROM AUTHOR]

Abstract In this article we give an overview over the use of DNP-enhanced solid-state NMR spectroscopy for the investigation of unfolded, disordered and misfolded proteins. We first provide an overview over studies in which DNP spectroscopy has successfully been applied for the structural investigation of well-folded amyloid fibrils formed by short peptides as well as full-length proteins. Sample cooling to cryogenic temperatures often leads to severe line broadening of resonance signals and thus a loss in resolution. However, inhomogeneous line broadening at low temperatures provides valuable information about residual dynamics and flexibility in proteins, and, in combination with appropriate selective isotope labeling techniques, inhomogeneous linewidths in disordered proteins or protein regions may be exploited for evaluation of conformational ensembles. In the last paragraph we highlight some recent studies where DNP-enhanced MAS-NMR-spectroscopy was applied to the study of disordered proteins/protein regions and inhomogeneous sample preparations. Graphical abstract Image 1 [ABSTRACT FROM AUTHOR]

Biological membranes define the interface of life and its basic unit, the cell. Membrane proteins play key roles in membrane functions, yet their structure and mechanisms remain poorly understood. Breakthroughs in crystallography and electron microscopy have invigorated structural analysis while failing to characterise key functional interactions with lipids, small molecules and membrane modulators, as well as their conformational polymorphism and dynamics. NMR is uniquely suited to resolving atomic environments within complex molecular assemblies and reporting on membrane organisation, protein structure, lipid and polysaccharide composition, conformational variations and molecular interactions. The main challenge in membrane protein studies at the atomic level remains the need for a membrane environment to support their fold. NMR studies in membrane mimetics and membranes of increasing complexity offer close to native environments for structural and molecular studies of membrane proteins. Solution NMR inherits high resolution from small molecule analysis, providing insights from detergent solubilised proteins and small molecular assemblies. Solid-state NMR achieves high resolution in membrane samples through fast sample spinning or sample alignment. Recent developments in dynamic nuclear polarisation NMR allow signal enhancement by orders of magnitude opening new opportunities for expanding the applications of NMR to studies of native membranes and whole cells.

The polarization enhancement of $\rm ^{19}F$ in $\rm LaF_{3}$ nanoparticles by dynamic nuclear polarization was investigated. A sample was prepared with nanoparticles that were produced by mechanical milling and dispersed in an ethanol matrix containing a free radical, 2,2,6,6,-Tetramethyl-1-piperidinyloxy. At a magnetic field of 2.5 T and temperature of 0.9 K, $\rm ^{19}F$ polarizations of $2.9\%$ and $-3.0\%$ were achieved. The observed polarizations are about 2/3 as high as those for $\rm ^{1}H$ in ethanol. This method can be applied for polarizing various nuclei in other materials, thus allowing a wide range of applications for polarized nuclear targets. [ABSTRACT FROM AUTHOR]

Complex mixtures, commonly encountered in metabolomics and food analytics, are now routinely measured by nuclear magnetic resonance (NMR) spectroscopy. Since many samples must be measured, one-dimensional proton (1D 1H) spectroscopy is the experiment of choice. A common challenge in complex mixture 1H NMR spectroscopy is spectral crowding, which limits the assignment of molecular components to those molecules in relatively high abundance. This limitation is exacerbated when the sample quantity itself is limited and concentrations are reduced even further during sample preparation for routine measurement. To address these challenges, we report a novel microfluidic NMR platform integrating signal enhancement via parahydrogen induced hyperpolarisation. The platform simultaneously addresses the challenges of handling small sample quantities through microfluidics, the associated decrease in signal given the reduced sample quantity by Signal Amplification by Reversible Exchange (SABRE), and overcoming spectral crowding by taking advantage of the chemosensing aspect of the SABRE effect. SABRE at the microscale is enabled by an integrated PDMS membrane alveolus, which provides bubble-free hydrogen gas contact with the sample solution. With this platform, we demonstrate high field NMR chemosensing of microliter sample volumes, nanoliter detection volumes, and micromolar concentrations corresponding to picomole molecular sensitivity. [ABSTRACT FROM AUTHOR]

Nuclear magnetic resonance (NMR) techniques play an essential role in natural science and medicine. In spite of the tremendous utility associated with the small energies detected, the most severe limitation is the low signal‐to‐noise ratio. Dynamic nuclear polarization (DNP), a technique based on transfer of polarization from electron to nuclear spins, has emerged as a tool to enhance sensitivity of NMR. However, the approach in liquids still faces several challenges. Herein we report the observation of room‐temperature, liquid DNP 13C signal enhancements in organic small molecules as high as 600 at 9.4 Tesla and 800 at 1.2 Tesla. A mechanistic investigation of the 13C‐DNP field dependence shows that DNP efficiency is raised by proper choice of the polarizing agent (paramagnetic center) and by halogen atoms as mediators of scalar hyperfine interaction. Observation of sizable DNP of 13CH2 and 13CH3 groups in organic molecules at 9.4 T opens perspective for a broader application of this method. [ABSTRACT FROM AUTHOR]

Abstract: The insufficient resolution of conventional methods has long limited the structural elucidation of cellulose and its derivatives, especially for those with relatively low crystallinities or in native cell walls. Recent 2D/3D solid-state NMR studies of 13C uniformly labeled plant biomaterials have initiated a re-investigation of our existing knowledge in cellulose structure and its interactions with matrix polymers but for unlabeled materials, this spectroscopic method becomes impractical due to limitations in sensitivity. Here, we investigate the molecular structure of unlabeled cotton cellulose by combining natural abundance 13C-13C 2D correlation solid-state NMR spectroscopy, as enabled by the sensitivity-enhancing technique of dynamic nuclear polarization, with statistical analysis of the observed and literature-reported chemical shifts. The atomic resolution allows us to monitor the loss of Iα and Iβ allomorphs and the generation of a novel structure during ball-milling, which reveals the importance of large crystallite size for maintaining the Iα and Iβ model structures. Partial order has been identified in the "disordered" domains, as evidenced by a discrete distribution of well-resolved peaks. This study not only provides heretofore unavailable high-resolution insights into cotton cellulose but also presents a widely applicable strategy for analyzing the structure of cellulose-rich materials without isotope-labeling. This work was part of a multi-technique study of ball-milled cotton described in the previous article in the same issue.Graphical abstract: [ABSTRACT FROM AUTHOR]

Since reaching the clinic, magnetic resonance imaging (MRI) has become an irreplaceable radiological tool because of the macroscopic information it provides across almost all organs and soft tissues within the human body, all without the need for ionising radiation. The sensitivity of MR, however, is too low to take full advantage of the rich chemical information contained in the MR signal. Hyperpolarisation techniques have recently emerged as methods to overcome the sensitivity limitations by enhancing the MR signal by many orders of magnitude compared to the thermal equilibrium, enabling a new class of metabolic and molecular X-nuclei based MR tracers capable of reporting on metabolic processes at the cellular level. These hyperpolarised (HP) tracers have the potential to elucidate the complex metabolic processes of many organs and pathologies, with studies so far focusing on the fields of oncology and cardiology. This review presents an overview of hyperpolarisation techniques that appear most promising for clinical use today, such as dissolution dynamic nuclear polarisation (d-DNP), parahydrogen-induced hyperpolarisation (PHIP), Brute force hyperpolarisation and spin-exchange optical pumping (SEOP), before discussing methods for tracer detection, emerging metabolic tracers and applications and progress in preclinical and clinical application. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • A frequency-agile 200 GHz/300 MHz DNP NMR spectrometer was constructed. • The spectrometer is based on solid-state mm-wave devices and quasioptical components. • 200 GHz photonic band gap resonator was integrated into double-tuned NMR saddle coil. • 13C DNP enhancement of ca. 1500 was obtained in synthetic diamond crystals at room temperature. • Resonator probehead provides the same electronic B 1 at up to 11 dB lower incident power. Abstract The most critical condition for performing Dynamic Nuclear Polarization (DNP) NMR experiments is achieving sufficiently high electronic B 1e fields over the sample at the matched EPR frequencies, which for modern high-resolution NMR instruments fall into the millimeter wave (mmW) range. Typically, mmWs are generated by powerful gyrotrons and/or extended interaction klystrons (EIKs) sources and then focused onto the sample by dielectric lenses. However, further development of DNP methods including new DNP pulse sequences may require B 1e fields higher than one could achieve with the current mmW technology. In order to address the challenge of significantly enhancing the mmW field at the sample, we have constructed and tested one-dimensional photonic band-gap (PBG) mmW resonator that was incorporated inside a double-tuned radiofrequency (rf) NMR saddle coil. The photonic crystal is formed by stacking ceramic discs with alternating high and low dielectric constants and thicknesses of λ/4 or 3λ/4, where λ is the wavelength of the incident mmW field in the corresponding dielectric material. When the mmW frequency is within the band gap of the photonic crystal, a defect created in the middle of the crystal confines the mmW energy, thus forming a resonant structure. An aluminum mirror in the middle of the defect has been used to substitute one-half of the structure with its mirror image in order to reduce the resonator size and simplify its tuning. The latter is achieved by adjusting the width of the defect by moving the aluminum mirror with respect to the dielectric stack using a gear mechanism. The 1D PBG resonator was the key element for constructing a multi-resonant integrated DNP/NMR probehead operating at 190–199 GHz EPR/300 MHz 1H/75.5 MHz 13C NMR frequencies. Initial tests of the multi-resonant DNP/NMR probehead were carried out using a quasioptical mmW bridge and a Bruker Biospin Avance II spectrometer equipped with a standard Bruker 7 T wide-bore 89 mm magnet parked at 300.13 MHz 1H NMR frequency. The mmW bridge built with all solid-state active components allows for the frequency tuning between ca. 190 and ca. 199 GHz with the output power up to 27 dBm (0.5 W) at 192 GHz and up to 23 dBm (0.2 W) at 197.5 GHz. Room temperature DNP experiments with a synthetic single crystal high-pressure high-temperature (HPHT) diamond (0.3 × 0.3 × 3.0 mm3) demonstrated dramatic 1500-fold enhancement of 13C natural abundance NMR signal at full incident mmW power. Significant 13C DNP enhancement (of about 90) have been obtained at incident mmW powers of as low as <100 μW. Further tests of the resonator performance have been carried out with a thin (ca. 100 μm thickness) composite polystyrene-microdiamond film by controlling the average mmW power at the optimal DNP conditions via a gated mode of operation. From these experiments, the PBG resonator with loaded Q ≃ 250 and finesse F ≈ 75 provides up to 12-fold or 11 db gain in the average mmW power vs. the non-resonant probehead configuration employing only a reflective mirror. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • A cryostat designed for dynamic nuclear polarization supports three probes. • Magic angle spinning experiments are performed at temperatures as low as 4.2 K. • Instrumentation development for dynamic nuclear polarization is described. Abstract Dynamic nuclear polarization (DNP) with cryogenic magic angle spinning (MAS) provides significant improvements in NMR sensitivity, yet presents unique technical challenges. Here we describe a custom cryostat and suite of NMR probes capable of manipulating nuclear spins with multi-resonant radiofrequency circuits, cryogenic spinning below 6 K, sample exchange, and microwave coupling for DNP. The corrugated waveguide and six transfer lines needed for DNP and cryogenic spinning functionality are coupled to the probe from the top of the magnet. Transfer lines are vacuum-jacketed and provide bearing and drive gas, variable temperature fluid, two exhaust pathways, and a sample ejection port. The cryostat thermally isolates the magnet bore, thereby protecting the magnet and increasing cryogen efficiency. This novel design supports cryogenic MAS-DNP performance over an array of probes without altering DNP functionality. We present three MAS probes (two supporting 3.2 mm rotors and one supporting 9.5 mm rotors) interfacing with the single cryostat. Mechanical details, transmission line radio frequency design, and performance of the cryostat and three probes are described. [ABSTRACT FROM AUTHOR]

Dynamic nuclear polarization (DNP) has enabled enormous gains in magnetic resonance signals and led to vastly accelerated NMR/MRI imaging and spectroscopy. Unlike conventional cw-techniques, DNP methods that exploit the full electron spectrum are appealing since they allow direct participation of all electrons in the hyperpolarization process. Such methods typically entail sweeps of microwave radiation over the broad electron linewidth to excite DNP but are often inefficient because the sweeps, constrained by adiabaticity requirements, are slow. In this paper, we develop a technique to overcome the DNP bottlenecks set by the slow sweeps, using a swept microwave frequency comb that increases the effective number of polarization transfer events while respecting adiabaticity constraints. This allows a multiplicative gain in DNP enhancement, scaling with the number of comb frequencies and limited only by the hyperfine-mediated electron linewidth. We demonstrate the technique for the optical hyperpolarization of 13C nuclei in powdered microdiamonds at low fields, increasing the DNP enhancement from 30 to 100 measured with respect to the thermal signal at 7T. For low concentrations of broad linewidth electron radicals [e.g., TEMPO ((2,2,6,6- tetramethylpiperidin-1-yl)oxyl)], these multiplicative gains could exceed an order of magnitude. [ABSTRACT FROM AUTHOR]

Gyrotrons have been developed as terahertz wave sources in dynamic nuclear polarisation (DNP) system to increase the sensitivity of nuclear magnetic resonance (NMR) in the high-magnetic field regime. However, the transmission of the Gaussian beam from the gyrotron output window to the NMR system proves to be a critical challenge. In this study, a terahertz transmission system, composed of a rectangular corrugated waveguide and a back-to-back rectangular corrugated horn for Gaussian mode filtering, is proposed. The attenuation in the optimised 0.33 THz corrugated waveguide remains lower than 0.01 dB/m in a broad frequency range from 280 GHz to 450 GHz. Besides, the coupling coefficient from the electric field in the 0.33 THz mode-filtering horn to the fundamental HE11 mode in the rectangular corrugated waveguide is obtained nearly 99.5%. This terahertz transmission system is potential to be applied in DNP-NMR spectrometer based on continuous wave gyrotrons. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • A 4-channel NMR probe and spectrometer with split excitation channel were developed. • Throughput for hyperpolarized NMR experiments was increased. • Protein-ligand binding affinity was determined from a single four-point experiment. Abstract Dissolution dynamic nuclear polarization (D-DNP) is a powerful technique to improve NMR sensitivity by a factor of thousands. Combining D-DNP with NMR-based screening enables to mitigate solubility or availability problems of ligands and target proteins in drug discovery as it can lower the concentration requirements into the sub-micromolar range. One of the challenges that D-DNP assisted NMR screening methods face for broad application, however, is a reduced throughput due to additional procedures and time required to create hyperpolarization. These requirements result in a delay of several tens of minutes in-between each NMR measurement. To solve this problem, we have developed a simultaneous 4-channel detection method for hyperpolarized 19F NMR, which can increase throughput fourfold by utilizing a purpose-built multiplexed NMR spectrometer and probe. With this system, the concentration-dependent binding interactions were observed for benzamidine and benzylamine with the serine protease trypsin. A T 2 relaxation measurement of a hyperpolarized reporter ligand (TFBC; CF 3 C 6 H 4 CNHNH 2), which competes for the same binding site on trypsin with the other ligands, was used. The hyperpolarized TFBC was mixed with trypsin and the ligand of interest, and injected into four flow cells inside the NMR probe. Across the set of four channels, a concentration gradient was created. From the simultaneously acquired relaxation datasets, it was possible to determine the dissociation constant (K D) of benzamidine and benzylamine without the requirement for individually optimizing experimental conditions for different affinities. A simulation showed that this 4-channel detection method applied to D-DNP NMR extends the screenable K D range to up to three orders of magnitude in a single experiment. [ABSTRACT FROM AUTHOR]

We introduce a novel design for millimeter wave electromagnetic structures within magic angle spinning (MAS) rotors. In this demonstration, a copper coating is vacuum deposited onto the outside surface of a sapphire rotor at a thickness of 50 nm. This thickness is sufficient to reflect 197‐GHz microwaves, yet not too thick as to interfere with radiofrequency fields at 300 MHz or prevent sample spinning due to eddy currents. Electromagnetic simulations of an idealized rotor geometry show a microwave quality factor of 148. MAS experiments with sample rotation frequencies of ωr/2π = 5.4 kHz demonstrate that the drag force due to eddy currents within the copper does not prevent sample spinning. Spectra of sodium acetate show resolved 13C J-couplings of 60 Hz and no appreciable broadening between coated and uncoated sapphire rotors, demonstrating that the copper coating does not prevent shimming and high-resolution nuclear magnetic resonance spectroscopy. Additionally, 13C Rabi nutation curves of ω1/2π = 103 kHz for both coated and uncoated rotors indicate no detrimental impact of the copper coating on radio frequency coupling of the nuclear spins to the sample coil. We present this metal coated rotor as a first step towards an MAS resonator. MAS resonators are expected to have a significant impact on developments in electron decoupling, pulsed dynamic nuclear polarization (DNP), room temperature DNP, DNP with low-power microwave sources, and electron paramagnetic resonance detection. [ABSTRACT FROM AUTHOR]