1H Magic Angle Spinning Nuclear Magnetic Resonance Research Articles

1H NMR spectroscopic characterisation of HepG2 cells as a model metabolic system for toxicology studies

The immortalised human hepatocellular HepG2 cell line is commonly used for toxicology studies as an alternative to animal testing due to its characteristic liver-distinctive functions. However, little is known about the baseline metabolic changes within these cells upon toxin exposure. We have applied 1H Nuclear Magnetic Resonance (NMR) spectroscopy to characterise the biochemical composition of HepG2 cells at baseline and post-exposure to hydrogen peroxide (H2O2). Metabolic profiles of live cells, cell extracts, and their spent media supernatants were obtained using 1H high-resolution magic angle spinning (HR-MAS) NMR and 1H NMR spectroscopic techniques. Orthogonal partial least squares discriminant analysis (O-PLS-DA) was used to characterise the metabolites that differed between the baseline and H2O2 treated groups. The results showed that H2O2 caused alterations to 10 metabolites, including acetate, glutamate, lipids, phosphocholine, and creatine in the live cells; 25 metabolites, including acetate, alanine, adenosine diphosphate (ADP), aspartate, citrate, creatine, glucose, glutamine, glutathione, and lactate in the cell extracts, and 22 metabolites, including acetate, alanine, formate, glucose, pyruvate, phenylalanine, threonine, tryptophan, tyrosine, and valine in the cell supernatants. At least 10 biochemical pathways associated with these metabolites were disrupted upon toxin exposure, including those involved in energy, lipid, and amino acid metabolism. Our findings illustrate the ability of NMR-based metabolic profiling of immortalised human cells to detect metabolic effects on central metabolism due to toxin exposure. The established data sets will enable more subtle biochemical changes in the HepG2 model cell system to be identified in future toxicity testing.

Molecular Profiling of Peanut under Raw, Roasting, and Autoclaving Conditions Using High-Resolution Magic Angle Spinning and Solution 1H NMR Spectroscopy.

Higher rates of peanut allergy have been observed in countries that commonly roast peanuts prior to consumption. Despite the importance of understanding the role of thermal processing in allergy and on peanut composition, studies toward generating signatures that identify molecular contents following processing are scant. Here, we identified spectral signatures to track changes and differences in the molecular composition of peanuts under raw, roasted, and high-pressure and high-temperature autoclaved conditions. We analyzed both the solid flesh of the seed and solutions derived from soaking peanuts using High-Resolution Magic Angle Spinning (HR-MAS) and solution 1H Nuclear Magnetic Resonance (NMR) spectroscopy, respectively. The NMR spectra of intact peanuts revealed triglycerides as the dominant species, assigned on the basis of multiplets at 4.1 and 4.3 ppm, and corresponding defatted flours revealed the presence of sugars. Sucrose assigned based on a doublet at 5.4 ppm (anomeric proton), and triglycerides were the most abundant small molecules observed, with little variation between conditions. Soaked peanut solutions were devoid of lipids, and their resulting spectra matched the profiles of defatted peanuts. Spectral signatures resulting from autoclaving differed strikingly between those from raw and roasted peanuts, with considerable line-broadening in regions corresponding to proteins and amino-acid side chains, from 0.5 to 2.0 ppm and 6.5 to 8.5 ppm. Taken together, by using complementary NMR methods to obtain a fingerprint of the molecular components in peanuts, we demonstrated that autoclaving led to a distinct composition, likely resulting from the hydrolytic cleavage of proteins, the most important molecule of the allergic reaction.

Magnetic Resonance-Based Analytical Tools to Study Polyvinylpyrrolidone-Hydroxyapatite Composites.

The synthesis of biocompatible and bioresorbable composite materials, such as a "polymer matrix-mineral constituent," stimulating the natural growth of living tissues and the restoration of damaged parts of the body, is one of the challenging problems in regenerative medicine and materials science. Composite films of bioresorbable polymer of polyvinylpyrrolidone (PVP) and hydroxyapatite (HA) were obtained. HA was synthesized in situ in the polymer solution. We applied electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) approaches to study the composite films' properties. The application of EPR in two frequency ranges allowed us to derive spectroscopic parameters of the nitrogen-based light and radiation-induced paramagnetic centers in HA, PVP and PVP-HA with high accuracy. It was shown that PVP did not significantly affect the EPR spectral and relaxation parameters of the radiation-induced paramagnetic centers in HA, while light-induced centers were detected only in PVP. Magic angle spinning (MAS) 1H NMR showed the presence of two signals at 4.7 ppm and -2.15 ppm, attributed to "free" water and hydroxyl groups, while the single line was attributed to 31P. NMR relaxation measurements for 1H and 31P showed that the relaxation decays were multicomponent processes that can be described by three components of the transverse relaxation times. The obtained results demonstrated that the applied magnetic resonance methods can be used for the quality control of PVP-HA composites and, potentially, for the development of analytical tools to follow the processes of sample treatment, resorption, and degradation.

Effects of glycerol content on structure and molecular motion in thermoplastic starch-based nanocomposites during long storage

Thermoplastic starch-based nanocomposites with varying glycerol content and montmorillonite as a nanofiller were studied using dynamic-mechanical analysis (DMA), X-ray diffraction (XRD) and nuclear magnetic resonance (NMR) during one-year storage. DMA results showed that starch-rich and glycerol-rich domains were present in the samples and during storage for up to one year the content of the amorphous phase decreased and molecular mobility changed. 13C NMR and XRD measurements confirmed that ordered structures were formed during storage and its content was larger for samples with higher glycerol content and increased with the storage time. The data obtained from deconvolutions of 1H broad line NMR spectra indicate increased overall molecular mobility in the samples up to four months of storage, while after nine months the trends were opposite. Lower free water content compared to the total water content in the samples determined according to deconvoluted 1H MAS (magic-angle spinning) NMR spectra indicated that a part of water molecules was immobilized in the ordered structures.

1H NMR Study of the HCa2Nb3O10 Photocatalyst with Different Hydration Levels.

The photocatalytic activity of layered perovskite-like oxides in water splitting reaction is dependent on the hydration level and species located in the interlayer slab: simple or complex cations as well as hydrogen-bonded or non-hydrogen-bonded H2O. To study proton localization and dynamics in the HCa2Nb3O10·yH2O photocatalyst with different hydration levels (hydrated—α-form, dehydrated—γ-form, and intermediate—β-form), complementary Nuclear Magnetic Resonance (NMR) techniques were applied. 1H Magic Angle Spinning NMR evidences the presence of different proton containing species in the interlayer slab depending on the hydration level. For α-form, HCa2Nb3O10·1.6H2O, 1H MAS NMR spectra reveal H3O+. Its molecular motion parameters were determined from 1H spin-lattice relaxation time in the rotating frame (T1ρ) using the Kohlrausch-Williams-Watts (KWW) correlation function with stretching exponent β = 0.28: eV, s. For the β-form, HCa2Nb3O10·0.8H2O, the only 1H NMR line is the result of an exchange between lattice and non-hydrogen-bonded water protons. T1ρ(1/T) indicates the presence of two characteristic points (224 and 176 K), at which proton dynamics change. The γ-form, HCa2Nb3O10·0.1H2O, contains bulk water and interlayer H+ in regular sites. 1H NMR spectra suggest two inequivalent cation positions. The parameters of the proton motion, found within the KWW model, are as follows: eV, s.

Compositional, structural, and surface characterization of heat-treated halloysite samples: Influence of surface treatment on photochemical activity

Quasi-homogeneous nanoparticulate halloysite clay with layered scroll morphology is an ideal material for photochemical activity due to the lack of interference from Fe contaminants and mixed mineral phases. In this work, halloysite was used as a model system for studying the influence of compositional defect sites on the structure and photochemical activity as a result of thermal modifications in the range of 400–480 °C and with 30–180 min exposure. The change in the Al-coordination environment was studied by 27Al, 1H magic-angle spinning (MAS) and cross-polarization (CP)/MAS nuclear magnetic resonance (NMR), and infrared (IR) spectroscopic techniques. The structural and morphological changes were followed by powder X-ray diffraction (XRD) and transmission electron microscope (TEM) techniques. Pore structure description was obtained from nitrogen gas adsorption measurements. The amount of residual hydroxide groups was determined by thermogravimetric analysis (TGA). Temperature-controlled desorption (TPD) and 1H MAS NMR techniques were used for the characterization of acid-base properties of thermally created defect sites. These defect sites showed a decrease in Al-coordination number and an increase in the number of acidic sites. However, evidence for the formation of basic sites during long exposure experiments can be seen, which emphasises the impact of the duration of the heat treatment. The emergence of acidic sites was correlated with the increase in photochemical activity. A significantly different photochemical activity was found for thermally treated halloysite than expected from the semiconductor-type photocatalysis model on the basis of the number of hydroxyl radicals generated as a function of the irradiation time.

Direct observation of the active sites in methane dehydroaromatization by NMR

Direct observation of the active sites in methane dehydroaromatization by NMR

Structural dynamics of CH3NH3+ and PbBr3\u2212 in tetragonal and cubic phases of CH3NH3PbBr3 hybrid perovskite by nuclear magnetic resonance

Understanding the structural dynamics of lead-halide perovskites is essential for their advanced use as photovoltaics.Here, the structural dynamics of the CH3NH3 cation and PbBr6 octahedra in the perovskite CH3NH3PbBr3 were studied via nuclear magnetic resonance (NMR) to determine the mechanism of the transition from the tetragonal to cubic phase. The chemical shifts were obtained by 1H, 13C, and 207Pb magic angle spinning NMR and 14N static NMR. The chemical shifts of the 1H nuclei in CH3 and NH3 remained constant with increasing temperature, whereas those of the 13C and 207Pb nuclei varied near the phase transition temperature (TC = 236 K), indicating that the structural environments of 13C and 207Pb change near TC. The spin–lattice relaxation time T1ρ values for 1H, 13C, and 207Pb nuclei increased with increasing temperature and did not exhibit an abrupt change near TC. In addition, the two lines in the 14N NMR spectra superposed into one line near TC, indicating the occurrence of a phase transition to a cubic phase with higher symmetry than tetragonal. Consequently, the main factor causing the phase transition from the tetragonal to cubic phase near TC is a change in the surroundings of the 207Pb nuclei in the PbBr6 octahedra and of the C–N groups in the CH3NH3 cations.

Spatially-resolved metabolic profiling of living Drosophila in neurodegenerative conditions using 1H magic angle spinning NMR

Drosophila flies are versatile animal models for the study of gene mutations in neuronal pathologies. Their small size allows performing in vivo Magic Angle Spinning (MAS) experiments to obtain high-resolution 1H nuclear magnetic resonance (NMR) spectra. Here, we use spatially-resolved 1H high-resolution MAS NMR to investigate in vivo metabolite contents in different segments of the fly body. A comparative study of metabolic changes was performed for three neurodegenerative disorders: two cell-specific neuronal and glial models of Huntington disease (HD) and a model of glutamate excitotoxicity. It is shown that these pathologies are characterized by specific and sometimes anatomically localized variations in metabolite concentrations. In two cases, the modifications of 1H MAS NMR spectra localized in fly heads were significant enough to allow the creation of a predictive model.

Assessing mesoscopic organization in copolymer-templated silica hybrid films via solid-state nuclear magnetic resonance

In the context of increasing use of nanostructured materials, finding innovative characterization methods able to assess precisely the level of ordering is essential. To this end, a range of model organic-inorganic copolymer-silica films is synthesized using poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) amphiphilic triblock copolymer as supramolecular template. Upon increasing copolymer concentration (10–100 wt%), the order can be gradually enhanced from short-range to long-range as proved by conventional techniques such as X-ray diffraction and electron microscopy. To evaluate the level of mesoscopic organization, the same series of samples is also analyzed by solid-state nuclear magnetic resonance (NMR) spectroscopy. The disorder-to-order transition is probed by 1H and 13C magic angle spinning NMR spectra through the increase in chemical environment uniformity and chain mobility respectively, which both result from the self-assembly mechanism. 1H NMR relaxation measurements (T2) using low-field NMR spectroscopy are instrumental to identify the threshold template concentration where ordering takes place.

Effects of sorbitol and formamide plasticizers on molecular motion in corn starch studied using NMR and DMTA

ABSTRACTMechanical and thermal properties of composite polymer materials strongly depend on their local structure and molecular dynamics which can be effectively studied by the solid‐state nuclear magnetic resonance (NMR) techniques. In the present paper, the influence of formamide (F) and sorbitol (S) plasticizers on molecular motion in thermoplastic starch (TPS) was studied using solid‐state NMR spectroscopy and dynamic mechanical thermal analysis (DMTA). DMTA measurements carried out for formamide‐ (F‐TPS) and sorbitol‐plasticized (S‐TPS) corn starches indicated heterogeneous plasticizer distribution of plasticizer‐rich and starch‐rich phases within the samples. The single pulse and cross‐polarization13C NMR spectra measured for both plasticized starches confirmed the amorphous character of their structure and distinctly different chain mobility supported by the values of13C spin–lattice relaxation times. The results of the analysis of broad line and magic angle spinning1H NMR spectra are in accordance with the results of DMTA measurements, revealing lower mobility of starch chains within S‐TPS in comparison to F‐TPS. Crosslinking of the starch chains with sorbitol molecules was suggested as being responsible for the lower mobility of the starch chains in S‐TPS. © 2020 Wiley Periodicals, Inc. J. Appl. Polym. Sci.2020,137, 48964.

Study on Paramagnetic Interactions of (CH3NH3)2CoBr4 Hybrid Perovskites Based on Nuclear Magnetic Resonance (NMR) Relaxation Time.

The thermal properties of organic–inorganic (CH3NH3)2CoBr4 crystals were investigated using differential scanning calorimetry and thermogravimetric analysis. The phase transition and partial decomposition temperatures were observed at 460 K and 572 K. Nuclear magnetic resonance (NMR) chemical shifts depend on the local field at the site of the resonating nucleus. In addition, temperature-dependent spin–lattice relaxation times (T1ρ) were measured using 1H and 13C magic angle spinning NMR to elucidate the paramagnetic interactions of the (CH3NH3)+ cations. The shortening of 1H and 13C T1ρ of the (CH3NH3)2CoBr4 crystals are due to the paramagnetic Co2+ effect. Moreover, the physical properties of (CH3NH3)2CoBr4 with paramagnetic ions and those of (CH3NH3)2CdBr4 without paramagnetic ions are reported and compared.

Capping Structure of Ligand-Cysteine on CdSe Magic-Sized Clusters.

Ligand molecules capping on clusters largely affect the formation and stabilization mechanism and the property of clusters. In semiconductor CdSe clusters, cysteine is used as one of the ligands and allows the formation of ultrastable (CdSe)34 magic-sized clusters. Cysteine has sulfhydryl, amine, and carboxylate groups, all of which have coordination ability to the CdSe surface, and the bonding states of the three functional groups of ligand–cysteine on the CdSe core have not been determined. In this work, the capping structure of ligand–cysteine is examined by performing Fourier transform infrared (FT-IR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and multinuclear solid-state nuclear magnetic resonance (NMR) spectroscopy. FT-IR, XPS, and 1H, 13C, and 23Na magic-angle spinning NMR show that the sulfhydryl group of ligand–cysteine forms a sulfur–cadmium bond with a cadmium atom at the CdSe surface, while the carboxylate group does not contribute to the protection of the CdSe core and binds to a sodium ion contained as a counterion. 15N–{77Se} through-bond J-single quantum filtered NMR experiment reveals that the amine group of ligand–cysteine has no coordination to selenium atoms. By considering the N–Cd bond forming ratio (∼43%) revealed in our previous work, which is confirmed in this work by analyzing 13Cα signal intensity (∼42%), we concluded that cysteine capping on (CdSe)34 occurs in two ways: one involves both the sulfur–cadmium and nitrogen–cadmium bonds, and the other bears only the sulfur–cadmium bond.

Hydration and Hydroxylation of MgO in Solution: NMR Identification of Proton-Containing Intermediate Phases.

Magnesium oxide (MgO)-engineered barriers used in subsurface applications will be exposed to high concentration brine environments and may form stable intermediate phases that can alter the effectiveness of the barrier. To explore the formation of these secondary intermediate phases, MgO was aged in water and three different brine solutions and characterized with X-ray diffraction (XRD) and 1H magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy. After aging, there is ∼4% molar equivalent of a hydrogen-containing species formed. The 1H MAS NMR spectra resolved multiple minor phases not visible in XRD, indicating that diverse disordered proton-containing environments are present in addition to crystalline Mg(OH)2 brucite. Density functional theory (DFT) simulations for the proposed Mg–O–H-, Mg–Cl–O–H-, and Na–O–H-containing phases were performed to index resonances observed in the experimental 1H MAS NMR spectra. Although the intermediate crystal structures exhibited overlapping 1H NMR resonances in the spectra, Mg–O–H intermediates were attributed to the growth of resonances in the δ +1.0 to 0.0 ppm region, and Mg–Cl–O–H structures produced the increasing contributions of the δ = +2.5 to 5.0 ppm resonances in the chloride-containing brines. Overall, 1H NMR analysis of aged MgO indicates the formation of a wide range of possible intermediate structures that cannot be observed or resolved in the XRD analysis.

NMR-Based Metabolomics in Cardiac Research.

Proton (1H) nuclear magnetic resonance (NMR) spectroscopy can measure a range of metabolites in both cardiac tissue and blood plasma for following cardiovascular disease. For solution-state NMR spectroscopy, it is necessary to create a tissue extract, with perchloric acid, acetonitrile/water, and chloroform/methanol being popular extraction media. Alternatively, high-resolution magic angle spinning (HRMAS) 1H NMR spectroscopy can be used to derive a metabolic profile directly from intact cardiac tissue. This chapter will discuss the practical methods used for 1H NMR spectroscopy to follow cardiovascular diseases both in terms of metabolic changes in cardiac tissue and changes in blood plasma.

Proton mobility in Ruddlesden–Popper phase H2La2Ti3O10 studied by 1H NMR

To study protons localization in H1.83K0.17La2Ti3O10·0.17H2O and their motional characteristics, complementary Nuclear Magnetic Resonance (NMR) techniques have been applied. 1H Magic Angle Spinning NMR evidences the presence of different proton containing species. By analyzing the temperature dependence of the 1H MAS NMR spectrum we attribute the observed lines to interlayer H+ in regular sites (isolated and in water rich environment), water protons and protons from various defects. The temperature behaviors of the spectral lines intensities and widths point out that intercalated water molecules are involved in translational motion that is confirmed by spin lattice relaxation rate (R1) and spin-lattice relaxation rate in rotating frame (R1ρ) measurements. It has been shown that for a correct determination of the proton motional parameters the Kohlrausch-Williams-Watts correlation function must be used. Its application results in the following parameters of proton motion in the interlayer space of H1.83K0.17La2Ti3O10·0.17H2O: Ea = 0.194(2) eV, β = 0.28(1), τ0=6.2(1)×10−10 s.

Revealing long- and short-range structural modifications within phosphorus-treated HZSM-5 zeolites by atom probe tomography, nuclear magnetic resonance and powder X-ray diffraction.

The average and the local structure of phosphorus-treated HZSM-5 zeolites were investigated by means of atom probe tomography, powder X-ray diffraction (at ambient and cryogenic temperatures) and 1H, 29Si, 27Al, and 31P magic angle spinning (MAS) solid state nuclear magnetic resonance (NMR) spectroscopy. Phosphatation to yield a product with P/Al ≤ 1 followed by thermal treatment leads to breaking of the Si-OH-Al bridging groups, and subsequent partial dealumination of the zeolite framework, as shown by the contraction of the orthorhombic unit-cell volume and by the loss of tetrahedral framework Al, as observed in the 27Al Multiple Quantum (MQ) MAS NMR spectrum. Most of the framework Al is present in an electronic environment distorted by the presence of phosphorus and appears not to be involved in classic Si-OH-Al Brønsted acid sites. The 31P MAS NMR signals indicate that phosphorus interacts with the zeolitic framework to locally form silico-aluminophosphate (SAPO) domains and the presence of a new kind of acidic site was confirmed by the resonance at ∼8.6 ppm in the 1H MAS NMR spectra, attributed to P-OH groups. Increasing the phosphorus loading (P/Al ≫ 1) promotes further dealumination of the framework and cross-dehydroxylation between P-OH and Si-OH species, leading to the formation of a crystalline silicon orthophosphate phase. With decreasing Al content, the monoclinic HZSM-5 structure becomes preferred, especially at 85 K where the strain relaxation is higher. However, the presence of a higher amount of silicophosphate impurities hinders the low-temperature strain release of the framework, indicating that some of these species are localized in the zeolite pores and contribute to the strain build up.

Nuclear Magnetic Resonance Study of O-ring Polymer Exposed to High-Pressure Hydrogen

Magic angle spinning (MAS) nuclear magnetic resonance (NMR) for <SUP>19</SUP>F and 1H nuclei was used how exposure to high-pressure (70 MPa) H₂ gas affected an O-ring material. <SUP>19</SUP>F MAS NMR revealed that poly(vinylidenefluoride-co-hexafluoroprophylene) in VDF/HFP = 76.1/23.9 mol % (FKM type 1) was involved in the O-ring material. This copolymer may undergo main-chain scission and termination to -CF₂H end groups during depressurization of high-pressure H₂. Evidence for formation of additional -CF₂H end group includes a small increase in the <SUP>19</SUP>F MAS NMR signal at -96 ppm, and a corresponding ¹H MAS NMR signal at 7.1 ppm in comparison with those in the material that had not been exposed to high-pressure H₂. Neither the <SUP>19</SUP>F nor the ¹H MAS NMR showed chemical shift after exposure to H₂; this absence of response means that exposure to H₂ did not affect the structure of the polymer.

Structural phase transitions and ferroelastic properties of perovskite-type layered (CH3NH3)2CdCl4

Herein, we perform a structural characterization of (CH3NH3)2CdCl4 by 1H and 13C magic-angle spinning (MAS) nuclear magnetic resonance (NMR) spectrometry, discussing the geometry around CH3NH3 cations. 1H MAS NMR determined two sets of protons in hydrogen-bonded CH3NH3 groups, exhibiting large T1ρ (L) and small T1ρ (S) values corresponding to long and short C-H/N-H bonds, respectively. The spin-lattice relaxation time of 113Cd in CdCl6 octahedra is shown to exhibit an anomaly near 283 K (TC2) due to the occurrence of a phase transition that is not governed by changes in the motion of CH3NH3 groups but is accompanied by the corresponding changes for Cd in CdCl6 units. The observed domain pattern corresponds to an exchange of a- and c-axes of the orthorhombic structure, with the resulting domain orientations viewed as the main reason for the occurrence of phase transitions, and changes in CdCl6 group motion observed by 113Cd NMR.

Fast MAS 1H NMR Study of Water Adsorption and Dissociation on the (100) Surface of Ceria Nanocubes: A Fully Hydroxylated, Hydrophobic Ceria Surface

1H nuclear magnetic resonance (NMR) spectroscopy was used to study hydroxylic surface species on ceria nanocubes, a crystalline, high-surface-area CeO2 that presents mostly (100) facets. Water adsorption and desorption experiments in combination with fast magic angle spinning (MAS, 20–40 kHz) 1H NMR provide high-resolution 1H spectra that allow the observation of ten resonance bands (water or hydroxyl) on or under the (100) surface. Assignments were made using a combination of adsorption and temperature-programmed desorption, quantitative spin counting, deuterium exchange, spin–lattice (T1) and spin–spin (T2) relaxation, and DFT calculations. In air, the (100) surface exists as a fully hydroxylated surface. Water adsorption and dissociation on dry ceria surfaces occur first at oxygen vacancies, but Ce3+ centers are not required since water dissociation is barrier-less on the fully oxidized surface. Surface −OH functionality occurs in two resolved bands representing isolated −OH (1 ppm) and hydrogen-bonded...