Heteronuclear Multiple Quantum Coherence Research Articles

High-resolution heteronuclear correlations between spin-1/2 and half-integer quadrupolar nuclei under fast MAS solid-state NMR

High isotropic resolution is essential for the structural elucidation of samples with multiple sites. In this study, utilizing the benefits of TRAPDOR-based heteronuclear multiple quantum coherence (T-HMQC) and a pair of one rotor period long cosine amplitude modulated low-power (cos-lp) pulse-based symmetric-split-t1 multiple-quantum magic angle spinning (MQMAS) methods, we have developed a proton-detected 2D 35Cl/1H T-HMQC-MQMAS pulse sequence under fast MAS (70 kHz) to achieve high-resolution in the indirect dimension of the spin-3/2 (35Cl) nuclei connected via protons. As T-HMQC polarizes not only single-quantum central transition (SQCT) but also triple-quantum (TQ) coherences, the proposed 2D pulse sequence is implemented via selection of two coherence pathways (SQCT→TQ →SQCT and TQ → SQCT→TQ) resulting in the 35Cl isotropic dimension and is superior to the existing double-quantum satellite-transition (DQST) T-HMQC in terms of resolution.

A new oxime synthesized from Senecio nutans SCh. Bip (chachacoma) reduces calcium influx in the vascular contractile response in rat aorta.

Senecio nutans Sch. Bip is an endemic plant commonly employed in the Andes culture to counteract the effects of mountain sickness, and its bioactive molecules could provide new drugs for treating hypertension. The purpose was to determine whether the vascular response of the plant bioactive molecules, such as (5-acetyl-6-hydroxy-2-isopropenyl-2,3-dihydrobenzofurane; Sn-I), could be improved by a simple structural modification to synthesize oximes (Ox-Sn-I). We characterized both compounds using IR and NMR spectroscopy and Heteronuclear Multiple Quantum Coherence (HMQC). We investigated vascular relaxation mechanisms in response to Sn-I and Ox-Sn-I using rat aorta and vascular smooth muscle cells (A7r5) as experimental models. Preincubation of aortic rings with Sn-I (10-5 M) significantly (p < 0.001) decreased the contractile effect in response to phenylephrine (PE) and potassium chloride (KCl). The sensitivity (EC50) to PE significantly (p < 0.01) decreased in the presence of Sn-I (10-5 M), but not with Ox-Sn-I. Sn-I significantly (p < 0.001) reduced the PE-induced contraction under calcium-free conditions. When A7r5 cells were preincubated with Sn-I and Ox-Sn-I (10-5 M), both compounds blunted the increase in intracellular Ca2+ induced by KCl. 2,3-Dihydrobenzofurane derived from S. nutans (Sn-I) reduces the contractile response probably by blocking Ca2+ entry through voltage-gated Ca2+ channels (VGCC) in vascular smooth cells. This effect also causes relaxation in rat aorta mediated by reduction of intracellular Ca2+ concentration, rather than an increase of NO generation in endothelial vascular cells.

AI-designed RF pulses enable fast pulsing heteronuclear multiple quantum coherence NMR experiment at high and ultra-high magnetic fields.

We present a new AI-optimized 2D heteronuclear multiple quantum coherence (RAPID-HMQC) pulse sequence for NMR spectroscopy. RAPID-HMQC is a longitudinal 1H relaxation-optimized experiment with new AI-designed band-selective pulses to accelerate the analysis of organic compounds, metabolites, biopolymers, and real-time monitoring of dynamic processes at high- and ultra-high magnetic fields.

Development of Low-Magnetic Susceptibility Microcoils via 5-Axis Machining for Analysis of Biological and Environmental Samples.

In environmental research, it is critical to understand how toxins impact invertebrate eggs and egg banks, which, due to their tiny size, are very challenging to study by conventional nuclear magnetic resonance (NMR) spectroscopy. Microcoil technology has been extensively utilized to enhance the mass-sensitivity of NMR. In a previous study, 5-axis computer numerical control (CNC) micromilling (shown to be a viable alternative to traditional microcoil production methods) was used to create a prototype copper slotted-tube resonator (STR). Despite the excellent limit of detection (LOD) of the resonator, the quality of the line shape was very poor due to the magnetic susceptibility of the copper resonator itself. This is best solved using magnetic susceptibility-matched materials. In this study, approaches are investigated that improve the susceptibility while retaining the versatility of coil milling. One method involves machining STRs from various copper/aluminum alloys, while the other involves machining ones from an aluminum 2011 alloy and electroplating them with copper. In all cases, combining copper and aluminum to produce resonators resulted in improved line shape and SNR compared to pure copper resonators due to their reduced magnetic susceptibility. However, the copper-plated aluminum resonators showed optimal performance from the devices tested. The enhanced LOD of these STRs allowed for the first 1H-13C heteronuclear multiple quantum coherence (HMQC) of a single intact 13C-labeled Daphnia magna egg (∼4 μg total biomass). This is a key step toward future screening programs that aim to elucidate the toxic processes in aquatic eggs.

Alkali-Activated Brick Aggregates as Industrial Valorized Wastes: Synthesis and Properties

In recent works, many industrial by-products were employed as solid precursors for the synthesis of alkali-activated binders and as alternatives to Portland cement for the immobilization of hazardous, toxic and nuclear wastes. Among industrial wastes, alkali-activated brick was found to be an interesting porous composite for removing very toxic heavy metals (Pb2+, Cd2+, Co2+) and radio-nuclides (Sr2+, Cs+, Rb+) from aqueous solutions. The starting material is very attractive due to the presence of metakaolinite as a geo-polymer precursor and silica for increasing material permeability and facilitating water filtration. The alkaline reaction gave rise to geo-polymerization followed by partial zeolitization. Elemental surface micro-analysis was performed by Scanning Electron Microscopy (SEM) equipped with an Energy-Dispersive X-ray Spectrometer (EDS). The formation of crystalline phases was corroborated by X-ray diffraction (XRD) analysis. Information about 29Si, 27Al and 1H nuclei environments in crystallized and amorphous aluminosilicates was obtained by 29Si, 27Al and 1H MAS NMR. 27Al–1H dipolar-mediated correlations were investigated by employing dipolar hetero-nuclear multiple quantum coherence (D-HMQC) NMR, highlighting Al–O–H bonds in bridging hydroxyl groups (Si–OH–Al) that are at the origin of adsorptive properties. Aqueous structural stability and cationic immobilization characteristics before and after material calcination were investigated from acid-leaching experiments.

Identification of hexahydrocannabinol (HHC), dihydro-iso-tetrahydrocannabinol (dihydro-iso-THC) and hexahydrocannabiphorol (HHCP) in electronic cigarette cartridge products.

Since 2021, products claiming to contain hexahydrocannabinol (HHC) and hexahydrocannabiphorol (HHCP), which are tetrahydrocannabinol (THC) analogs, have been distributed via the Internet. Owing to the presence of three asymmetric carbons in their structure, HHC and HHCP have multiple stereoisomers. This study aimed to identify the actual stereoisomers of HHC and HHCP isolated from electronic cigarette cartridge products using nuclear magnetic resonance (NMR) spectroscopy. Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-photodiode array-mass spectrometry (LC-PDA-MS) were used for the analyses of two major peaks and one minor peak in product A and two major peaks in product B. These five compounds were isolated by silica gel column chromatography, and their structures were analyzed by 1H, 13C-NMR and various two-dimensional NMR techniques, i.e., H-H correlation spectroscopy, heteronuclear multiple quantum coherence, heteronuclear multiple-bond correlation, and nuclear Overhauser effect spectroscopy. Three compounds isolated from product A were identified as rel-(6aR,9R,10aR)-hexahydrocannabinol (11β-hexahydrocannabinol; 11β-HHC), rel-(6aR,9S,10aR)-hexahydrocannabinol (11α-hexahydrocannabinol, 11α-HHC), and a minor compound (2R,5S,6R)-dihydro-iso-tetrahydrocannabinol (dihydro-iso-THC). Meanwhile, the structural isomers of the major compound isolated from product B were identified as rel-(6aR, 9R, 10aR)-hexahydrocannabiphorol (11β-hexahydrocannabiphorol; 11β-HHCP) and rel-(6aR, 9S, 10aR)-hexahydrocannabiphorol (11α-hexahydrocannabiphorol; 11α-HHCP). The presence of both 11β-HHC and 11α-HHC in the HHC products analyzed in this study suggests that they were most likely synthesized via the reduction reaction of Δ8-THC or Δ9-THC. Dihydro-iso-THC was probably obtained as a byproduct of the synthesis of Δ8-THC or Δ9-THC from cannabidiol. Similarly, 11β-HHCP and 11α-HHCP in the HHCP product could stem from Δ9-tetrahydrocannabiphorol.

Comparison of methods for 14N-1H recoupling in 14N-1H HMQC MAS NMR

1H-detected 14N heteronuclear multiple-quantum coherence (HMQC) magic-angle-spinning (MAS) NMR experiments performed at fast magic-angle spinning (≥50 kHz) are finding increasing application, e.g., to pharmaceuticals. Of importance to the efficacy of these techniques is the recoupling technique applied to reintroduce the 1H-14N dipolar coupling. In this paper, we compare, by experiment and 2-spin density matrix simulations, two classes of recoupling scheme: first, those based on n = 2 rotary resonance, namely R3 and spin-polarisation inversion SPI-R3, and the symmetry based SR412 method and, second, the TRAPDOR method. Both classes require optimisation depending on the magnitude of the quadrupolar interaction, and thus there is a compromise choice for samples with more than one nitrogen site, as is the case for the studied dipeptide β-AspAla that contains two nitrogen sites with a small and large quadrupolar coupling constant. Considering this, we observe better sensitivity for the TRAPDOR method, though noting the marked sensitivity of TRAPDOR to the 14N transmitter offset, with both SPI-R3 and SR412 giving similar recoupling performance.

Advanced Solid-State NMR Analysis of Two Crystal Forms of Ranitidine Hydrochloride: Detection of 1H-14N Intra-/Intermolecular Correlations.

Understanding the characteristics of crystal polymorphism of active pharmaceutical ingredients and analyzing them with high sensitivity is important for quality of drug products, appropriate characterization strategies, and appropriate screening and selection processes. However, there are few methods to measure intra- and intermolecular correlations in crystals other than X-ray crystallography, for which it is sometimes difficult to obtain suitable single crystals. Recently, solid-state NMR has been recognized as a straightforward method for measuring molecular correlations. In this study, we selected ranitidine hydrochloride, which is known to exist in two forms, 1 and 2, as the model drug and investigated each form using solid-state NMR. In conducting the analysis, rotating the sample tube, which had a 1-mm inner diameter, increased the solid-state NMR resolution at 70 kHz. The 1H-14N dipolar-based heteronuclear multiple quantum coherence (D-HMQC) analysis revealed the intermolecular correlation of Form 1 between the N atom of the nitro group and a proton of the furan moiety, which were closer than those of the intramolecular correlation reported using single X-ray crystal analysis. Thus, 1H-14N D-HMQC analysis could be useful for characterizing intermolecular interaction in ranitidine hydrochloride crystals. In addition, we reassigned the 13C solid-state NMR signals of ranitidine hydrochloride according to the liquid-state and multiple solid-state NMR experiments.

1H and 13C NMR Study of Oligosuccinimide Prepared by Thermal Condensation and Evaluation of Its Scale Inhibition

&lt;p&gt;The presence of scale is a serious problem in the petroleum industry, and some efforts should be made to control scale formation. Oligosuccinimide (OSI) is an oligomer with several repeating units of succinimide. The structure of the OSI main chain and end groups has never been comprehensively analyzed. Meanwhile, OSI is potentially a scale inhibitor due to chelating properties, high polarity, and high solubility. This study investigates the molecular structure of oligosuccinimide, focusing on the main chain and end groups. Synthesis was carried out by thermal condensation between maleic anhydride and ammonium carbonate using a &lt;sup&gt;1&lt;/sup&gt;H and &lt;sup&gt;13&lt;/sup&gt;C NMR spectroscope equipped with Distortionless Enhancement by Polarization Transfer 135, Heteronuclear Multiple Quantum Coherence, and Single Quantum Coherence. The NMR analysis results detected the presence of the main chain and several synthesized OSI end groups, such as the amino, succinimide, and maleimide end groups. However, the dicarboxylic acid end group and other irregular structures, as in polysuccinimide (various synthesis methods), were not identified. It confirmed that our thermal condensation method produced OSI with less irregular structures than previous methods. Evaluation of OSI as CaCO&lt;sub&gt;3&lt;/sub&gt; and CaSO&lt;sub&gt;4&lt;/sub&gt; scales inhibitor showed reasonably good performance in very small concentrations. The inhibition efficiency was 73.20% for the CaCO&lt;sub&gt;3&lt;/sub&gt; scale with 10 mg.L&lt;sup&gt;-1&lt;/sup&gt; and 55.29% for the CaSO&lt;sub&gt;4&lt;/sub&gt; scale with 10 mg.L&lt;sup&gt;-1&lt;/sup&gt; inhibitor concentration. Analysis of thermal stability informed OSI has good thermal stability because it started to be degraded at 353.38 °C.&lt;/p&gt;

Improved resolution for spin-3/2 isotopes in solids via the indirect NMR detection of triple-quantum coherences using the T-HMQC sequence

The indirect NMR detection of quadrupolar nuclei in solids under magic-angle spinning (MAS) is possible with the through-space HMQC (heteronuclear multiple-quantum coherence) scheme incorporating the TRAPDOR (transfer of population in double-resonance) dipolar recoupling. This sequence, called T-HMQC, exhibits limited t1-noise. In this contribution, with the help of numerical simulations of spin dynamics, we show that most of the time, the fastest coherence transfer in the T-HMQC scheme is achieved when TRAPDOR recoupling employs the highest radiofrequency (rf) field compatible with the probe specifications. We also demonstrate how the indirect detection of the triple-quantum (3Q) coherences of spin-3/2 quadrupolar nuclei in solids improves the spectral resolution for these isotopes. The sequence is then called T-HMQC3. We demonstrate the gain in resolution provided by this sequence for the indirect proton detection of 35Cl nuclei in l-histidine∙HCl and l-cysteine∙HCl, as well as that of 23Na isotope in NaH2PO4. These experiments indicate that the gain in resolution depends on the relative values of the chemical and quadrupolar-induced shifts (QIS) for the different spin-3/2 species. In the case of NaH2PO4, we show that the transfer efficiency of the T-HMQC3 sequence employing an rf-field of 80 kHz with a MAS frequency of 62.5 kHz reaches 75% of that of the t1-noise eliminated (TONE) dipolar-mediated HMQC (D-HMQC) scheme.

Characterization of kraft lignin prepared from mixed hardwoods by 2D HMQC and 31P NMR analyses

This study was conducted to determine the lignin substructures, hydroxyl (phenolic + aliphatic) contents, and carboxyl contents in kraft lignin (KL) prepared from mixed hardwoods by using 2D Heteronuclear Multiple Quantum Coherence Nuclear Magnetic Resonance (HMQC NMR) and 31P NMR techniques. Based on 2D HMQC NMR analysis of KL, stilbene and vanillin substructures were present in the aromatic region, while trace amounts of β-O-4 and β-β moieties were detected in the oxygenated aliphatic region. The total hydroxyl content calculated from 31P NMR was 5.24 mmol/g KL. The aliphatic hydroxyl content was 1.04 mmol/g KL, and phenolic hydroxyl content was 4.20 mmol/g KL. Of the phenolic hydroxyl groups, the contribution of syringyl (S) and guaiacyl (G) was 1.02 and 0.97 mmol/g KL, respectively. The S/G molar ratio of KL calculated from 31P NMR was 1.05. The carboxyl content was 0.44 mmol/g KL.

19F fast MAS (60–111 kHz) dipolar and scalar based correlation spectroscopy of organic molecules and pharmaceutical formulations

19F magic angle spinning (MAS) NMR spectroscopy is a powerful tool for characterization of fluorinated solids. The recent development of 19F MAS NMR probes, operating at spinning frequencies of 60–111 kHz, enabled analysis of systems spanning from organic molecules to pharmaceutical formulations to biological assemblies, with unprecedented resolution. Herein, we systematically evaluate the benefits of high MAS frequencies (60–111 kHz) for 1D and 2D 19F-detected experiments in two pharmaceuticals, the antimalarial drug mefloquine and a formulation of the cholesterol-lowering drug atorvastatin calcium. We demonstrate that 1H decoupling is essential and that scalar-based, heteronuclear single quantum coherence (HSQC) and heteronuclear multiple quantum coherence (HMQC) correlation experiments become feasible and efficient at the MAS frequency of 100 kHz. This study opens doors for the applications of high frequency 19F MAS NMR to a wide range of problems in chemistry and biology.

Local Structure and Protons in Non-Stoichiometric Pseudo-Cubic Pollucite Mineral by Multinuclear NMR

The pollucite structure is considered as a candidate ceramic crystalline matrix for the ceramic immobilization and long-term storage of 135Cs and 137Cs fission products, and thus, their structural characteristics have particular importance. However, its local structure has not been fully resolved from reciprocal-space techniques and infrared spectroscopy, and important discrepancies exist in the available literature. Two birefringent and non-stoichiometric pollucite specimens from Tanco pegmatite (Cs0.83Na0.20Al1.13Si2.56O6) and from Mt. Mica pegmatite (Cs0.94Na018Al1.23Si2.78O6), with powder X-ray diffraction patterns fully consistent with the cubic Ia-3d space-group symmetry, and with a very different degree of hydrothermal alteration, were used in this work. High-resolution magic-angle spinning multinuclear magnetic resonance (MAS NMR) spectroscopy, including 29Si, 27Al, 23Na, 133Cs, and 1H spectra at 9.4 T, as well as 1H, 27Al, 27Al{1H} dipolar evolutions and 27Al{29Si} Heteronuclear Multiple Quantum Coherence (HMCQ) spectra at 17.6 T, has been used to investigate the local structure of pollucite and the role of protons. The 29Si spectra suggest a local structure with a disordered Si/Al distribution in only one tetrahedral T site, but with a preference of Si atoms for Q41 (3Si,1Al) and Q42 (2Si,2Al) environments, in comparison with random and Loewenstein distributions, due to charge dispersion effects. However, the 27Al{1H} dipolar evolutions suggest two spectroscopically distinct T sites for Al atoms. The 23Na and 133Cs spectra indicate broad site distributions for these cavity cations. The anisotropic character of the long-range disordered pollucite structure, with a pseudo-cubic symmetry and lack of strict periodicity, can be explained from an incipient displacive transition to lower symmetry. These pollucite specimens are essentially anhydrous minerals despite the 1H and the cross-polarization experiments suggesting that some protons exist in the structure as -OH groups, whereas water molecules were only found in relation to the phyllosilicate impurities from alteration in specimen Tanco and perhaps also as liquid water in fluid inclusions.

Molecular and Electronic Structure of Isolated Platinum Sites Enabled by the Expedient Measurement of 195Pt Chemical Shift Anisotropy.

Techniques that can characterize the molecular structures of dilute surface species are required to facilitate the rational synthesis and improvement of Pt-based heterogeneous catalysts. 195Pt solid-state NMR spectroscopy could be an ideal tool for this task because 195Pt isotropic chemical shifts and chemical shift anisotropy (CSA) are highly sensitive probes of the local chemical environment and electronic structure. However, the characterization of Pt surface-sites is complicated by the typical low Pt loadings that are between 0.2 and 5 wt% and broadening of 195Pt solid-state NMR spectra by CSA. Here, we introduce a set of solid-state NMR methods that exploit fast MAS and indirect detection using a sensitive spy nucleus (1H or 31P) to enable the rapid acquisition of 195Pt MAS NMR spectra. We demonstrate that high-resolution wideline 195Pt MAS NMR spectra can be acquired in minutes to a few hours for a series of molecular and single-site Pt species grafted on silica with Pt loading of only 3-5 wt%. Low-power, long-duration, sideband-selective excitation, and saturation pulses are incorporated into t1-noise eliminated dipolar heteronuclear multiple quantum coherence, perfect echo resonance echo saturation pulse double resonance, or J-resolved pulse sequences. The complete 195Pt MAS NMR spectrum is then reconstructed by recording a series of 1D NMR spectra where the offset of the 195Pt pulses is varied in increments of the MAS frequency. Analysis of the 195Pt MAS NMR spectra yields the 195Pt chemical shift tensor parameters. Zeroth order approximation density functional theory calculations accurately predict 195Pt CS tensor parameters. Simple and predictive orbital models relate the CS tensor parameters to the Pt electronic structure and coordination environment. The methodology developed here paves the way for the detailed structural and electronic analysis of dilute platinum surface-sites.

Quantitative NMR analysis of the kinetics of prenucleation oligomerization and aggregation of pathogenic huntingtin exon-1 protein

The N-terminal region of the huntingtin protein, encoded by exon-1 (httex1) and containing an expanded polyglutamine tract, forms fibrils that accumulate in neuronal inclusion bodies, resulting in Huntington's disease. We previously showed that reversible formation of a sparsely populated tetramer of the N-terminal amphiphilic domain, comprising a dimer of dimers in a four-helix bundle configuration, occurs on the microsecond timescale and is an essential prerequisite for subsequent nucleation and fibril formation that takes place orders of magnitude slower on a timescale of hours. For pathogenic httex1, such as httex1Q35 with 35 glutamines, NMR signals decay too rapidly to permit measurement of time-intensive exchange-based experiments. Here, we show that quantitative analysis of both the kinetics and mechanism of prenucleation tetramerization and aggregation can be obtained simultaneously from a series of 1H-15N band-selective optimized flip-angle short-transient heteronuclear multiple quantum coherence (SOFAST-HMQC) correlation spectra. The equilibria and kinetics of tetramerization are derived from the time dependence of the 15N chemical shifts and 1H-15N cross-peak volume/intensity ratios, while the kinetics of irreversible fibril formation are afforded by the decay curves of 1H-15N cross-peak intensities and volumes. Analysis of data on httex1Q35 over a series of concentrations ranging from 200 to 750 μM and containing variable (7 to 20%) amounts of the Met7O sulfoxide species, which does not tetramerize, shows that aggregation of native httex1Q35 proceeds via fourth-order primary nucleation, consistent with the critical role of prenucleation tetramerization, coupled with first-order secondary nucleation. The Met7O sulfoxide species does not nucleate but is still incorporated into fibrils by elongation.

Combining heteronuclear correlation NMR with spin-diffusion to detect relayed Cl–H–H and N–H–H proximities in molecular solids

Analysis of short-to-intermediate range intermolecular interactions offers a great way of characterizing the solid-state organization of small molecules and materials. This can be achieved by two-dimensional (2D) homo- and heteronuclear correlation NMR spectroscopy, for example, by carrying out experiments at high magnetic fields in conjunction with fast magic-angle spinning (MAS) techniques. But, detecting 2D peaks for heteronuclear dipolar coupled spin pairs separated by greater than 3 Å is not always straightforward, particularly when low-gamma quadrupolar nuclei are involved. Here, we present a 2D correlation NMR experiment that combines the advantages of heteronuclear-multiple quantum coherence (HMQC) and proton-based spin-diffusion (SD) pulse sequences using radio-frequency-driven-recouping (RFDR) to probe inter and intramolecular 1H-X (X = 14N, 35Cl) interactions. This experiment can be used to acquire 2D 1H{X}-HMQC filtered 1H–1H correlation as well as 2D 1H-X HMQC spectra. Powder forms of dopamine·HCl and l-histidine·HCl·H2O are characterized at high fields (21.1 T and 18.8 T) with fast MAS (60 kHz) using the 2D HMQC-SD-RFDR approach. Solid-state NMR results are complemented with NMR crystallography analyses using the gauge-including projector augmented wave (GIPAW) approach. For histidine·HCl·H2O, 2D peaks associated with 14N–1H–1H and 35Cl–1H–1H distances of up to 4.4 and 3.9 Å have been detected. This is further corroborated by the observation of 2D peaks corresponding to 14N–1H–1H and 35Cl–1H–1H distances of up to 4.2 and 3.7 Å in dopamine·HCl, indicating the suitability of the HMQC-SD-RFDR experiments for detecting medium-range proximities in molecular solids.

Novel Oxime Synthesized from a Natural Product of Senecio nutans SCh. Bip. (Asteraceae) Enhances Vascular Relaxation in Rats by an Endothelium-Independent Mechanism.

Senecio nutans Sch. Bip. and its constituents are reported to have antihypertensive effects. We isolated metabolite–1, a natural compound from S. nutans (4-hydroxy-3-(isopenten-2-yl)-acetophenone), and synthesized novel oxime – 1 (4-hydroxy-3-(isopenten-2-yl)-acetophenoxime) to evaluate their effect on vascular reactivity. Compounds were purified (metabolite–1) or synthetized (oxime–1) and characterized using IR and NMR spectroscopy and Heteronuclear Multiple Quantum Coherence (HMQC). Using pharmacological agents such as phenylephrine (PE) and KCl (enhancing contraction), acetylcholine (ACh), L-NAME (nitric oxide (NO) and endothelial function), Bay K8644-induced CaV1.2 channel (calcium channel modulator), and isolated aortic rings in an organ bath setup, the possible mechanisms of vascular action were determined. Pre-incubation of aortic rings with 10−5 M oxime–1 significantly (p < 0.001) decreased the contractile response to 30 mM KCl. EC50 to KCl significantly (p < 0.01) increased in the presence of oxime–1 (37.72 ± 2.10 mM) compared to that obtained under control conditions (22.37 ± 1.40 mM). Oxime–1 significantly reduced (p < 0.001) the contractile response to different concentrations of PE (10−7 to 10−5 M) by a mechanism that decreases Cav1.2-mediated Ca2+ influx from the extracellular space and reduces Ca2+ release from intracellular stores. At a submaximal concentration (10−5 M), oxime–1 caused a significant relaxation in rat aorta even without vascular endothelium or after pre-incubate the tissue with L-NAME. Oxime–1 decreases the contractile response to PE by blunting the release of Ca2+ from intracellular stores and blocking of Ca2+ influx by channels. Metabolite–1 reduces the contractile response to KCl, apparently by reducing the plasma membrane depolarization and Ca2+ influx from the extracellular space. These acetophenone derivates from S. nutans (metabolite–1 and oxime–1) cause vasorelaxation through pathways involving an increase of the endothelial NO generation or a higher bioavailability, further highlighting that structural modification of naturally occurring metabolites can enhance their intended pharmacological functions.

Revisiting Edge Sites of γ-Al2O3 Using Needle-Shaped Nanocrystals and Recoupling-Time-Encoded {27Al}-1H D-HMQC NMR Spectroscopy

Despite being widely used in numerous catalytic applications, our understanding of reactive surface sites of high-surface-area γ-Al2O3 remains limited to date. Recent contributions have pointed toward the potential role of highly reactive edge sites contained in the high-field signal (−0.5 to 0 ppm) of the 1H NMR spectrum of γ-Al2O3 materials. This work combines the development of well-defined, needle-shaped γ-Al2O3 nanocrystals having a high relative fraction of edge sites with the use of state-of-the-art solid-state NMR to significantly deepen our understanding of this specific signal. We are able to resolve two hydroxyl sites with distinct isotropic chemical shifts of −0.2 and −0.4 ppm and different positions within the dipole–dipole network from 1H–1H single-quantum double-quantum NMR. Moreover, the use of recoupling-time-encoded arbitrary-indirect-dwell dipolar heteronuclear multiple quantum coherence allows us to partially revise previous assignments for surface-aluminum sites in the proximity of these hydroxyl sites. Although previous work has ascribed the high-field signal to be correlated with a single four-coordinate Al-site with a substantial quadrupolar broadening of >10 MHz, we can identify the presence of two four-coordinate Al-sites with similar isotropic chemical shifts but different quadrupolar coupling constants of approximately 7 and >10 MHz, respectively. Recoupling-time-encoded data are thus able to differentiate sites that would otherwise only be achievable with access to multiple fields or usage of highly advanced NMR techniques.

Selective 1H-13C heteronuclear multiple quantum coherence experiments for 1H-13C pairs near 19F

Variants of the heteronuclear multiple quantum coherence (HMQC) experiment are presented in which only 1H-13C pairs that interact with a 19F nucleus report. One version of this experiment selects for scalar coupling between the 1H and 19F nucleus (dubbed the SELFPRO-J-HMQC), the other for nuclear Overhauser effect-based interactions between 1H and 19F (SELFPRO-nOe-HMQC). The experiment can be executed such that only directly bound 1H-13C pairs report, or such that longer range 1H-13C correlations report as well. A selective shaped pulse in 19F was inserted in the SELFPRO-J-HMQC; in this version, only 1H-13C pairs that interact with the single chosen 19F nucleus will report. Example spectra are shown of trifluorotoluene and two more complex pharmaceutical molecules.

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

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