Intermolecular Single-quantum Coherences Research Articles

Tracking Metabolite Variations during the Degradation of Vegetables in Rice Bran Bed with Intact-State Nuclear Magnetic Resonance Spectroscopy.

Fermentation-a process of compound degradation by microorganisms-is a traditional food processing method utilized worldwide for the long-term preservation of fresh foods. In recent years, fermented foods have gained attention as health foods. Fermentation increases the nutritional value of ingredients, producing complex flavors and aromas. To identify unknown components in fermented foods, it is necessary to analyze compounds and conditions nondestructively and comprehensively. We performed intact-state nuclear magnetic resonance (NMR) spectroscopy using intermolecular single quantum coherence (iSQC) to detect the degradation of vegetables directly and nondestructively. We used two types of vegetables and a rice bran bed (nukazuke), which is used for traditional vegetable fermentation in Japan. Major metabolites such as saccharides, organic acids, and amino acids were identified in iSQC-sliced spectra. Comparing NMR signal intensities during degradation revealed the transition of metabolites characteristic of lactic acid fermentation. A pathway-based network analysis showed pathways involved in amino acid metabolism and lactic acid fermentation. Our analytical approach with intact-state NMR spectroscopy using iSQC demonstrated that it may be effective in other experimental systems, allowing for the evaluation of phenomena that have been conventionally overlooked in their true state.

Exploring Proton-Only NMR Experiments and Filters for Daphnia In Vivo: Potential and Limitations.

Environmental metabolomics provides insight into how anthropogenic activities have an impact on the health of an organism at the molecular level. Within this field, in vivo NMR stands out as a powerful tool for monitoring real-time changes in an organism's metabolome. Typically, these studies use 2D 13C-1H experiments on 13C-enriched organisms. Daphnia are the most studied species, given their widespread use in toxicity testing. However, with COVID-19 and other geopolitical factors, the cost of isotope enrichment increased ~6-7 fold over the last two years, making 13C-enriched cultures difficult to maintain. Thus, it is essential to revisit proton-only in vivo NMR and ask, "Can any metabolic information be obtained from Daphnia using proton-only experiments?". Two samples are considered here: living and whole reswollen organisms. A range of filters are tested, including relaxation, lipid suppression, multiple-quantum, J-coupling suppression, 2D 1H-1H experiments, selective experiments, and those exploiting intermolecular single-quantum coherence. While most filters improve the ex vivo spectra, only the most complex filters succeed in vivo. If non-enriched organisms must be used, then, DREAMTIME is recommended for targeted monitoring, while IP-iSQC was the only experiment that allowed non-targeted metabolite identification in vivo. This paper is critically important as it documents not just the experiments that succeed in vivo but also those that fail and demonstrates first-hand the difficulties associated with proton-only in vivo NMR.

In‐Phase Ultra High‐Resolution In Vivo NMR

AbstractAlthough current NMR techniques allow organisms to be studied in vivo, magnetic susceptibility distortions, which arise from inhomogeneous distributions of chemical moieties, prevent the acquisition of high‐resolution NMR spectra. Intermolecular single quantum coherence (iSQC) is a technique that breaks the sample's spatial isotropy to form long range dipolar couplings, which can be exploited to extract chemical shift information free of perturbations. While this approach holds vast potential, present practical limitations include radiation damping, relaxation losses, and non‐phase sensitive data. Herein, these drawbacks are addressed, and a new technique termed in‐phase iSQC (IP‐iSQC) is introduced. When applied to a living system, high‐resolution NMR spectra, nearly identical to a buffer extract, are obtained. The ability to look inside an organism and extract a high‐resolution metabolic profile is profound and should find applications in fields in which metabolism or in vivo processes are of interest.

In-Phase Ultra High-Resolution In Vivo NMR.

Although current NMR techniques allow organisms to be studied in vivo, magnetic susceptibility distortions, which arise from inhomogeneous distributions of chemical moieties, prevent the acquisition of high-resolution NMR spectra. Intermolecular single quantum coherence (iSQC) is a technique that breaks the sample's spatial isotropy to form long range dipolar couplings, which can be exploited to extract chemical shift information free of perturbations. While this approach holds vast potential, present practical limitations include radiation damping, relaxation losses, and non-phase sensitive data. Herein, these drawbacks are addressed, and a new technique termed in-phase iSQC (IP-iSQC) is introduced. When applied to a living system, high-resolution NMR spectra, nearly identical to a buffer extract, are obtained. The ability to look inside an organism and extract a high-resolution metabolic profile is profound and should find applications in fields in which metabolism or in vivo processes are of interest.

A heteronuclear intermolecular single-quantum coherence scheme for high-resolution 2D J-resolved 1H NMR spectra in inhomogeneous magnetic fields

ABSTRACTBased on heteronuclear intermolecular single-quantum coherences between proton (1H) and quadrupolar nuclei (i.e. deuterium 2H), a three-dimensional nuclear magnetic resonance (NMR) pulse sequence is proposed for recovering high-resolution two-dimensional J-resolved NMR spectra from samples mixed with a deuterated solvent in the presence of large magnetic field inhomogeneities. Benefitting from excitation of spins via two different radio frequency (RF) transmit channels, this sequence is suitable for applications in randomly large inhomogeneous fields and the solvent suppression generally required in homonuclear intermolecular multiple-quantum coherence approaches is no longer necessary. Systematic theoretical analyses are given based on the distant dipolar field treatment. Experiment on a sample of corn oil in deuterated acetone and ethyl 3-bromopropionate and acetone dissolved in DMSO-d6 in a deshimmed field with severe inhomogeneous broadening is performed to show the feasibility and applicability of this sequence.

Usage of the ultrafast intermolecular single-quantum coherence (UF iSQC) sequence for NMR spectroscopy of ex vivo tissue

High-resolution nuclear magnetic resonance (NMR) spectroscopy has been considered as a powerful tool for noninvasive, rapid, and accurate composition analyses on solution samples. However, because tissues are in semisolid phase, the high-resolution spectra cannot be obtained by conventional NMR spectroscopy. In the present study, an ultrafast intermolecular single quantum coherence (UF iSQC) NMR spectroscopic method based on spatial encoding is used for direct analyses of intact salmon muscle tissues, intact eggs from shishamo smelt, and a whole zebra fish. Most of free amino acid and fatty acid signals can be directly detected within 5min in their original state without tedious sample preparation processes. The experimental results show that not only can the UF iSQC method overcome the inhomogeneity of biological tissues, but also it can greatly reduce experimental times owing to the introduction of ultrafast acquisition schemes. Changes occurring in amino acids and fatty acids of salmon muscle tissues, eggs from shishamo smelt, and the whole zebra fish during storage are investigated. Similar variation patterns of metabolic profiles have been deduced for three fish samples, yielding good agreements with literature results.

Spatially-encoded intermolecular single-quantum coherence method for high-resolution NMR spectra in inhomogeneous fields

A spatially-encoded intermolecular single-quantum coherence pulse sequence is utilized to obtain high-resolution NMR spectra in inhomogeneous fields. To enhance sensitivities, a soft pulse was employed to solely excite solvent spins. Only inhomogeneous field information is exhibited along the indirect dimension, and spectral-width narrowing in the indirect dimension not only enhances the spectral sensitivity, but also increases measurable spectral widths in the direct dimension. The sequence shortens the acquisition time from minutes to seconds. A detailed theoretical analysis is provided. The sensitivity and the resolution of the method are proved by the studies of chemical and biological materials.

Feasibility of Ultrafast Intermolecular Single-Quantum Coherence Spectroscopy in Analysis of Viscous-Liquid Foods

A newly developed nuclear magnetic resonance (NMR) technique, ultrafast intermolecular single-quantum coherence (UF iSQC) spectroscopy, demonstrated its potentials to obtain spectra of viscous-liquid samples in deuterium-free environment. UF iSQC not only inherits the advantage of ultrafast acquisition efficiency of spatial encoding but also keeps the intrinsic merit of immunity of iSQC in inhomogeneous magnetic field. In the present study, UF iSQC spectroscopy is used for the direct analysis of three typical viscous-liquid foods, namely, honey, yogurt, and tomato sauce, and its feasibility is verified by ultrafast acquisition of NMR spectra of these viscous-liquid foods. UF iSQC spectroscopy presents an advantageous alternative to directly detect most components of these samples in favorable spectral information within 1 min, thus saving the trouble of complicated sample pretreatments and tedious shimming processes. Due to the immunity to field inhomogeneity, the UF iSQC technique can be a proper supplement to dilution, extraction NMR, and magic angle spinning. In addition, in comparison with the conventional iSQC method, the acquisition time of the proposed method is greatly reduced due to the introduction of the spatial encoding technique. The lower sensitivity and resolution of UF iSOC spectra than those of dilutions acquired by the conventional NMR method could be improved by some newly developed strategies. Results obtained here demonstrate the potential application of the UF iSQC spectroscopy in the quality control of food.

Hadamard-encoded high-resolution NMR spectroscopy via intermolecular single-quantum coherences

NMR spectroscopy plays an important role in metabolite studies because it can provide atomic level information critical for understanding biological systems. Nevertheless, NMR investigations on biological tissues are hampered by the magnetic field inhomogeneities originating from variations in macroscopic magnetic susceptibility, which lead to broad spectral lines and subsequently obscure metabolite signals. A new pulse sequence based on intermolecular single-quantum coherences was proposed to obtain one-dimensional high-resolution NMR spectra in inhomogeneous magnetic fields via Hadamard encoding. The new method can provide resolution-improved spectra directly through one-dimensional acquisition within a relatively short acquisition time. Theoretical derivation was performed and the conclusion was tested by solution samples in purposely de-shimmed magnetic fields and pig brain tissue sample. The experimental results show that this sequence can yield useful structural information, even when the field inhomogeneity is sufficiently severe to erase almost all spectral information with conventional one-dimensional single-quantum coherence techniques. Moreover, good solvent suppression efficiency can be achieved by this sequence. This sequence may provide a promising way for high-resolution NMR spectroscopy of biological tissue.

In vivo spatially localized high resolution 1H MRS via intermolecular single‐quantum coherence of rat brain at 7 T

To compare the conventional localized point-resolved spectroscopy (PRESS) with localized 2D intermolecular single-quantum coherence (iSQC) magnetic resonance spectroscopy (MRS) and obtain in vivo MRS spectrum of rat brain using the latter technique. A brain phantom, an intact pig brain tissue, and mature Sprague-Dawley rat were studied by PRESS, Nano magic-angle spinning spectroscopy, and iSQC MRS. Using PRESS, high-resolution MRS can be obtained from the brain phantom and pig brain tissue with a small voxel in a relatively homogeneous field. When a large voxel is selected, the field homogeneity is distinctly reduced. No useful information is obtained from the PRESS spectra. However, using the iSQC MRS, high-resolution spectra can be obtained from the two samples with a relatively large voxel. In the same way, an iSQC MRS spectrum can be obtained from a relatively large voxel of in vivo rat brain with a comparable resolution to the PRESS spectrum with a small voxel. Compared to PRESS, the iSQC MRS may be more feasible and promising for detection of strongly structured tissues with relatively large voxels.

An Intermolecular Single-Quantum Coherence Detection Scheme for High-Resolution Two-Dimensional J-resolved Spectroscopy in Inhomogeneous Fields

A new pulse sequence based on intermolecular single-quantum coherences (iSQCs) is proposed to achieve high-resolution two-dimensional (2D) J-resolved spectra in inhomogeneous fields via three-dimensional (3D) acquisition. Since the iSQC evolution period and spin echo evolution period in this sequence are intrinsically insensitive to magnetic field inhomogeneities, high-resolution 2D J-resolved spectra can be recovered from nuclei in inhomogeneous fields by projecting the 3D data onto the 2D plane. Analytical expressions of the resulting signals were derived assuming the secular dipole-dipole interaction. Analyses of a solution sample placed in a deliberately unshimmed magnetic field and of a biological sample with intrinsic field inhomogeneities were performed. The results show that this sequence provides an attractive and efficient way to eliminate the influence of field inhomogeneities on 2D J-resolved spectra, which is potentially useful for characterizing complex chemical materials and studying biological metabolites in inhomogeneous fields.

Intermolecular single-quantum coherence sequences for high-resolution NMR spectra in inhomogeneous fields

A new pulse sequence based on intermolecular single-quantum coherences (iSQCs) is proposed to obtain high-resolution NMR spectroscopy in inhomogeneous magnetic fields via fast 2D acquisition. Taking the intrinsic properties of iSQCs, the sequence is time-efficient with a narrow spectral width in the indirect dimension. It can recover useful information of chemical shifts, relative peak areas, J coupling constants, and multiplet patterns even when the field inhomogeneity is severe enough to erase almost all spectroscopic information. Moreover, good solvent suppression efficiency can be achieved by this sequence even with imperfect radio-frequency pulse flip angles. Spatially localized iSQC spectroscopy was performed on a sample packed with pig brain tissue and cucumber to show the feasibility of the sequence in in vivo magnetic resonance spectroscopy (MRS). This sequence may provide a promising way for the applications on in vivo and in situ high-resolution NMR spectroscopy.

Heteronuclear intermolecular single-quantum coherences in liquid nuclear magnetic resonance

This paper analyses the heteronuclear Cosy Revamped by Asymmetric Z-gradient Echo Detection pulse sequence. General theoretical expressions of the pulse sequence with arbitrary flip angles were derived by using dipolar field treatment and signals originating from heteronuclear intermolecular single-quantum coherences (iSQCs) in highly-polarized two spin-1/2 systems were mainly discussed in order to find the optimal flip angles. The results show that signals from heteronuclear iSQCs decay slower than those from intermolecular double-quantum coherences or intermolecular zero-quantum coherences. Magical angle experiments validate that the signals are from heteronuclear iSQCs and insensitive to the imperfection of radio-frequency flip angles. All experimental observations are in excellent agreement with theoretical predictions. The quantum-mechanical treatment leads to similar predictions to the dipolar field treatment.

Double-quantum-filtered intermolecular single-quantum coherences in nuclear magnetic resonance spectroscopy and imaging

Based on the double-quantum-filtered technique, a three-pulse sequence was designed to effectively eliminate contamination of residual conventional single-quantum coherences (SQC) from intermolecular SQC signals in single-resonance spin-1/2 samples such as water. Analytical expressions were derived from the modified Bloch equations with dipolar fields. All experimental observations and numerical simulation results are in excellent agreement with the theoretical predictions. Compared to the CRAZED sequence, the sequence can effectively suppress residual conventional SQC signals even with great imperfection of radio-frequency pulse flip angles. For the first time, signals from pure intermolecular SQCs were selectively observed in forms of spectrum or image.

Sensitivity to local dipole fields in the CRAZED experiment: An approach to bright spot MRI

Local dipole fields such as those created by small iron-oxide particles are used to produce regions of low intensity (dark contrast) in many molecular magnetic resonance imaging applications. We have investigated, with computer simulations and experiments at 17.6 T, how the COSY revamped with asymmetric z-gradient echo detection (CRAZED) experiment that selects intermolecular double-quantum coherences can also be used to visualize such local dipole fields. Application of the coherence-selection gradient pulses parallel to the main magnetic field produced similar, dark contrast as conventional gradient echo imaging. Application of the gradient along the magic angle leads to total loss of signal intensity in homogeneous samples. In the presence of local dipole fields, the contrast was inverted and bright signals from the dipoles were observed over a very low background. Both simulations and experiments showed that the signal strongly decreased when a phase-cycle suppressing single-quantum coherences was employed. Therefore, we conclude that most of the signal comes from directly refocused magnetization or intermolecular single-quantum coherences. Finally, we demonstrate that bright contrast from local dipole fields can also be obtained, when the pair of coherence-selection gradient pulses is deliberately mismatched. Both methods allowed visualization of local dipole fields in phantoms in experimental times of about 3 min.

Observation and characterization of intermolecular homonuclear single-quantum coherences in liquid nuclear magnetic resonance

Intermolecular zero-quantum and double-quantum coherences (iZQCs and iDQCs) are frequently discussed in literature since they may provide novel contrast mechanisms in magnetic resonance imaging and possibilities for high-resolution spectra in an inhomogeneous and unstable magnetic field. In a previous paper [J. Chem. Phys. 115, 10769 (2001)], we have studied both theoretically and experimentally the properties of iZQC and iDQC nuclear magnetic resonance (NMR) signals related to intermolecular dipolar interactions in two-component systems. In this paper, the investigation is extended to homonuclear intermolecular single-quantum coherences (iSQCs) from the second-order spin interactions, which have not been observed and studied previously. Selective excitation was used to suppress the strong conventional single-spin single-quantum signals. A combination of dipolar field treatment and Torrey equation was used to derive a general theoretical expression for the time evolution of spins with arbitrary flip angles of rf pulses. The expression was used to predict the optimal conditions for iSQCs among highly polarized spins in liquid. Dependence of the iSQC signals on the experimental parameters was measured and analyzed to verify the theoretical predictions. For the first time, signals from pure homonuclear two-spin iSQCs free of much larger conventional single-spin single-quantum signals, and intermolecular iSQC cross peaks in homonuclear pulsed-field gradient COSY experiments were observed and characterized, in one- and two-dimensional (1D and 2D) experiments, respectively. The use of coherence-selection gradients tilted at the magic angle results in the suppression of iSQC cross peaks. It provides strong evidence that the observed signals originate from distant dipolar interactions. Relaxation and diffusion properties of iSQCs in multiple-component samples were characterized and analyzed as well as the optimal rf flip angles. Theoretical and experimental results presented herein demonstrate that the signals from the homonuclear second-order iSQCs not only have a similar signal intensity as iZQCs or iDQCs, all of which are much stronger than that from three-spin iSQCs reported previously, but also provide spatial information related to dipolar correlation scales similar to iZQCs and iDQCs, which is not present in conventional SQC experiments. All 1D and 2D NMR experimental observations based on single- and multiple-component samples are in excellent agreement with the theoretical predictions. The quantitative study of iSQCs provides a better understanding of their unique mechanisms, and may find useful applications in NMR analyses such as sample purification and/or preparation of metabolites, biofluids, and natural compounds dissolved in nondeuterated solvents.