Two-dimensional Nuclear Magnetic Resonance Spectra Research Articles

Relaxation Editing in NMR Using a New Two-Dimensional Long-Lived Coherence Method for Mixture Analysis.

The exploration of metabolomics and targeted segments of proteins stands as a pivotal facet of Nuclear Magnetic Resonance (NMR) analysis, furnishing valuable insights into molecular architectures and potential therapeutic applications. The issue of spectral congestion frequently presents challenges in ascribing distinct peaks within the confines of both one-dimensional (1D) and two-dimensional (2D) NMR spectra. Numerous strategies have been proposed to resolve specific resonances in NMR spectra differentially. Among these approaches, relaxation editing emerges as a viable solution. In the realm of relaxation phenomena within NMR, Long-Lived States (LLS) and Long-Lived Coherences (LLC) manifest as promising phenomena, offering enhanced relaxation lifetimes in comparison to the traditional longitudinal (T1) and transverse (T2) relaxation times for coupled nuclear spins. Notably, LLC presents a pathway to attenuate uncoupled high-intensity peaks, effectively diminishing their impact. The foundation of this technique rests upon the premise that the relaxation lifetime in the rotating frame (T1ρ) remains smaller than TLLC. In pursuit of refining spectral assignments within complex mixtures, we introduce a new pulse sequence tailored for LLC Total Correlation Spectroscopy (LLC-TOCSY). This demonstrates efficacy in extracting LLC signals within configurations involving multiple coupled spins, thereby decluttering the spectrum and enhancing the accuracy of peak assignments. To validate the effectiveness of this method, a collection of samples was subjected to scrutiny, yielding compelling results.

Antimycobacterial and nitric oxide production inhibitory activities of limonoids isolated from Trichilia lepidota subsp. schumanniana (Harms) T.D.Penn

Six previously undescribed intact limonoids together with four known compounds were isolated from the seeds of Trichilia lepidota subsp. schumanniana (Harms) T.D.Penn. Their structures were characterized based on one- and two-dimensional nuclear magnetic resonance spectra, infrared, ultraviolet, mass spectroscopy results, and optical rotation. All compounds were evaluated for their ability to inhibit nitric oxide production in cultures of RAW 264.7 macrophages stimulated by lipopolysaccharide, cytotoxicity and growth of Mycobacterium tuberculosis strains H37Rv and M299. The compounds 7-deacetyl-11β,12α-diacetoxy-14,15-epoxyazadirone (5) and walsurin E (9) were the most potent in inhibiting nitric oxide production, although the compounds 1-deshydroxy-12α-acetoxymunronin N (1) and 6α,12α-dihydroxyazadirone (6) also showed controlled potential of this mediator, in addition to being potent growth inhibitors of Mycobacterium tuberculosis H37RV and M299, without cytotoxicity interference. Ring intact limonoids isolated from Trichilia lepidota subsp. schumanniana seeds are a new source of bioactive substances that may be used in the future against diseases such as tuberculosis and other processes related to inflammation.

Probing the Binding Mechanism of Acylated Peptides to Human Serum Albumin

Peptides represent an increasingly important class of pharmaceutical products. During the last decade or so, acylation with fatty acids has demonstrated considerable success in prolonging the circulating half-life of therapeutic peptides by exploiting the ability of fatty acids to reversibly bind to human serum albumin (HSA), thus significantly impacting their pharmacological profiles. Employing methyl-13C-labeled oleic acid or palmitic acid as probe molecules and exploiting HSA mutants designed to probe fatty acid binding, the signals in two-dimensional (2D) nuclear magnetic resonance (NMR) spectra corresponding to high-affinity fatty acid binding sites in HSA were assigned. Subsequently, using a set of selected acylated peptides, competitive displacement experiments by 2D NMR identified a primary fatty acid binding site in HSA utilized in acylated peptide binding. These results represent an important first step toward understanding the structural basis for acylated peptides binding to HSA.

Enhanced catalytic cleavage of C-O and C-C bonds of raw biomass into lignin monomers and glucose

Reductive catalytic fractionation (RCF) of lignocellulose has been considered to be a promising strategy for the separation and subsequent upgradation of biomass constituents. However, the recalcitrant C-C bonds in lignin were the major limitation to produce aromatic monomers in high yield for the hydrogenolysis of lignocellulosic biomass under mild conditions. Herein, an efficient approach was developed to catalytic in-situ hydrogenolysis of solid biomass into lignin monomers and glucose using Ni-Al catalyst with Ni2Al3 alloy phase. 62.9 wt% of aromatic monomers yield was achieved with the degree of delignification up to 89.8 %. Two-dimensional heteronuclear single quantum correlation nuclear magnetic resonance (2D HSQC NMR) spectrum emerged that the C-O and C-C bonds involving β-O-4, β-β and β-5 linkages were all catalytically broken. Moreover, the cellulose was recovered with the cellulose retention and purity of 80.1 % and 88.2 %, respectively, which was further enzymatically hydrolyzed to glucose with the yield of 92 %. Overall, the efficient fractionation and transformation of biomass constituents were realized simultaneously.

A Strategy for Pinpointing Natural Bioactive Components Using Two-Dimensional Bioassay Profilings Combined with Comprehensive Two-Dimensional Countercurrent Chromatography × High-Performance Liquid Chromatography.

Inspired by the interpretation of two-dimensional (2D) nuclear magnetic resonance spectra, an efficient strategy was proposed for pinpointing bioactive components from complex natural products. An off-line comprehensive countercurrent chromatography (CCC) × high-performance liquid chromatography (HPLC) was employed to achieve a 2D chemical chromatogram, and 2D bioassay profilings were obtained from bioassays of the eluent of the first dimension (1D) CCC and the eluent of the second dimension (2D) HPLC. Then 2D chemical chromatograms and 2D bioassay profilings were matched for pinpointing bioactive natural components from complex matrices. Thus, bioactive components in a complex matrix could be efficiently analyzed, separated, and bioactivity-determined. This experimental scheme was successfully demonstrated with a traditional medicinal herb Polygonum cuspidatum Sieb. et Zucc. The feasibility of this 2D strategy was verified with tyrosinase inhibition assay, α-glucosidase inhibition assay, DPPH radical scavenging assay, and ABTS•+ decolorization assay. Eight natural inhibitors were successfully pinpointed and identified from P. cuspidatum. Both pieceid-2″-O-gallate (10) and vanicoside B (20) were screened and identified as natural tyrosinase inhibitors for the first time. Meanwhile, vanicoside B (20) was also found as the strongest α-glucosidase inhibitor among all the isolated components. Most of the compounds exhibited much higher radical scavenging activities. Compared with traditional methodology based on one-dimensional chromatographic separation, the present 2D strategy would be more precise, efficient, and convenient to screen and separate bioactive compounds from complex matrices.

Investigation on the Mechanism of Radical Intermediate Formation and Moderate Oxidation of Spiro-OMeTAD by the Synergistic Effect of Multisubstituted Polyoxometalates in Perovskite Solar Cells.

Conventional oxidation of 2,2',7,7'-tetrakis[N,N-di(4-methoxyphenyl)amino]-9,9'-spirobifluorene (Spiro-OMeTAD) by air would bring various drawbacks for perovskite solar cells (PSCs), such as low power conversion efficiency (PCE) and poor stability. Here, a series of heteroatom-substituted Keggin-type polyoxometalates (POMs), H4PMo11VO40 (PMo11V), H5PMo10V2O40 (PMo10V2), and H6PMo9V3O40 (PMo9V3) are prepared and applied as p-type dopants to realize quantitative and controllable oxidation of Spiro-OMeTAD under an inert condition. The possible mechanism and electron donor regions in the oxidation of Spiro-OMeTAD are investigated using two-dimensional nuclear magnetic resonance (NMR) spectra and the relationship between POM structures and the oxidation degree of Spiro-OMeTAD is proposed. In addition, the synergistic effect of heteroatoms makes V2-substituted PMo10V2 exhibit appropriate oxidation of Spiro-OMeTAD and promoted the highest efficient hole extraction as well as the decreased charge recombination. Therefore, the champion device doped with PMo10V2 shows a PCE of 20.41% and a superior open circuit voltage (Voc) of 1.133 V, surpassing that of the pristine device (18.61%). This work presents a fresh perspective to the controllable oxidation of Spiro-OMeTAD employing economical inorganic POM dopants, which would promote the commercialization of PSCs.

Advances and trends in analytical techniques in natural product research: Challenges and future perspective

Advances in analytical methods and bioassay development have helped to push forward the research in natural products. Their high chemical diversity and the effects of evolutionary pressure to create biologically active molecules could be attributed to success in drug discovery. Despite the availability of modern analytical instrumentation techniques, metabolic profiling covers the identification of a selected group of metabolites. The phytochemical analysis is commonly performed using standard techniques such as thin-layer chromatography, high-performance thin-layer chromatography, high-performance liquid chromatography, gas chromatography and more recently mass spectrometry and nuclear magnetic resonance spectrometry. Two-dimensional J-resolved Nuclear Magnetic Resonance (NMR) spectra and multivariate data analysis techniques were applied to avoid low resolution and overlapping signals hampering the identification of the individual components of botanicals. On-targeted metabolomic analysis via the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF-MS) was utilised to localise compounds belonging to various chemical classes (i.e. oxygenated fatty acids, flavonols, phenolic acids, and sinapoyl choline derivatives). Rotation planar chromatography over-pressured layer chromatography and electro planar chromatography are the other innovations. Parallel and serially coupled layers open up new avenues for the analysis of a large number of samples for high-throughput screening and very complex matrices in a natural product. Analytical strategies with applications to natural extracts and novel methods that have strong potential, regardless of how often they are used are discussed with respect to their potential applications challenges and future trends.

Nightshift: A Python program for plotting simulated NMR spectra from assigned chemical shifts from the Biological Magnetic Resonance Data Bank.

Nuclear magnetic resonance (NMR) provides site specific information on local environments through chemical shifts. NMR is widely used in the study of proteins, ranging from determination of three-dimensional (3D) structures to characterizing dynamics and binding of small molecules and other proteins or ligands. Assigned chemical shift data for the atoms within proteins is a treasure trove of information that can facilitate a broad range of biochemical and biophysical studies. The Biological Magnetic Resonance Data Bank (BMRB) is a publicly accessible database that contains a large number of assigned chemical shifts; however, translating this wealth of knowledge into a practical application is not straightforward. Herein we present nightshift: a Python command line utility and library for plotting simulated two-dimensional (2D) and 3D NMR spectra from assigned chemical shifts in the BMRB. This tool allows users to simulate routinely collected amide and methyl fingerprint spectra, backbone triple-resonance assignment spectra, and user-defined custom correlations, including ones that do not necessarily correspond to published experiments. This tool enables experienced NMR spectroscopists, those learning the craft, and interested scientists seeking to utilize NMR the ability to preview or examine a wide range of spectra for proteins whose assignments are deposited in the BMRB, irrespective of whether those experiments have been executed or reported. The tool applies equally to folded and intrinsically disordered proteins, limited only by the existence of a BMRB deposition. The features of nightshift are described along with applications that illustrate the ease with which complicated correlation spectra and binding events can be simulated.

DEEP picker is a deep neural network for accurate deconvolution of complex two-dimensional NMR spectra

The analysis of nuclear magnetic resonance (NMR) spectra for the comprehensive and unambiguous identification and characterization of peaks is a difficult, but critically important step in all NMR analyses of complex biological molecular systems. Here, we introduce DEEP Picker, a deep neural network (DNN)-based approach for peak picking and spectral deconvolution which semi-automates the analysis of two-dimensional NMR spectra. DEEP Picker includes 8 hidden convolutional layers and was trained on a large number of synthetic spectra of known composition with variable degrees of crowdedness. We show that our method is able to correctly identify overlapping peaks, including ones that are challenging for expert spectroscopists and existing computational methods alike. We demonstrate the utility of DEEP Picker on NMR spectra of folded and intrinsically disordered proteins as well as a complex metabolomics mixture, and show how it provides access to valuable NMR information. DEEP Picker should facilitate the semi-automation and standardization of protocols for better consistency and sharing of results within the scientific community.

Conjugated Polyene Ketones From the Marine Sponge Clathria (Thalysias) Reinwardti (Vosmaer, 1880) and Their Cytotoxic Activity

Five conjugated polyene ketones (1-5) were isolated from the methanol extract of the marine sponge Clathria ( Thalysias) reinwardti (Vosmaer, 1880) living in the coastal waters of Vietnam. Their structures were determined to be 8-(2′,3′,4′-trimethylphenyl)-6-methyl-oct-3( E),5( E),7( E)-trien-2-one (1), 13-apoastaxanthinone (2), 9-apoastaxanthinone (3), 2,3-dehydro-4-oxo- β-ionone (4), and 4-(2′,3′,4′-trimethylphenyl)-but-3( E)-en-2-one (5), by extensive analysis of high-resolution electron spray ionization mass spectrum (HR-ESI-MS), one-dimensional, and two-dimensional (2D) nuclear magnetic resonance (NMR) spectra, as well as by comparison of the spectral data with those reported in the literature. Compound 1 was new, compounds 2 to 4 were isolated from nature for the first time, and the chemical structure as well as the NMR assignments, of 5 were indicated by 2D NMR for the first time. Additionally, compound 5 exhibited cytotoxic activity against the human cancer cells SK-LU-1, SK-Mel-2, MCF-7, and Hep-G2 with half-minimal inhibitory concentration (IC50) values of 15.12 ± 3.43, 17.41 ± 2.83, 33.12 ± 3.39, and 34.38 ± 3.52 µM, respectively, but displayed only a weak cytotoxic effect on the normal HEK-239A cells (IC50 64.67 ± 3.67 µM). Compound 5 also significantly increased Caspase-3 activity in SK-LU-1 cells at concentrations of 10, 15, and 20 µM.

Dynamics of uniformly labelled solid proteins between 100 and 300 K: A 2D 2H-13C MAS NMR approach

We describe a 2H based MAS nuclear magnetic resonance (NMR) method to obtain site-specific molecular dynamics of biomolecules. The method utilizes the use of deuterium nucleus as a spin label that is proven to be very useful in dynamics studies of solid biological and functional materials. The aim is to understand overall characteristics of protein backbone and side-chain motions for CD3, CD2 and CD groups, in terms of timescale, type and activation energy of the underlying processes. Variable temperature two-dimensional (2D) 2H-13C correlation MAS NMR spectra were recorded for the uniformly 2H,13C,15N labelled Alanine and microcrystalline SH3 at a broad temperature range, from 320 K down to 100 K. First, the deuterium quadrupolar-coupling constant from specific D-C sites is obtained with the 2D experiment by utilizing carbon chemical shifts. Second, the static quadrupolar patterns are obtained at 100 K. Third, variable temperature approach enabled the observation of quadrupolar pattern over different motional regimes; slow, intermediate and fast. And finally, the apparent activation energies for C-D sites are determined and compared, by evaluating the temperature induced signal intensities. This information led to the determination of the dynamic processes for different D-C sites at a broad range of temperature and motional timescales. This is a first representation of 2D 2H-13C MAS NMR approach applied to fully isotope labelled deuterated protein covering 220 K temperature range.

Physicochemical aspects of zwitterionic core-shell tecto dendrimers characterized by a thorough NMR investigation

Core-shell tecto dendrimers (CSTDs) are a class of superstructured dendrimeric nanoconstructs with similar structural characteristics and biomedical performances to the high-generation dendrimers that can be decorated with zwitterions to be availed of good antifouling properties for different biomedical applications. Characterization of the CSTD-based dendrimeric derivatives remains to be a great challenge. Here, we attempted to utilize nuclear magnetic resonance (NMR) technologies to investigate the structure and hydrodynamic size of CSTDs functionalized with zwitterionic carboxybetaine acrylamide (CBAA) and confirmed their antifouling property through protein resistance assay. In this study, we initially prepared CSTDs by supramolecular assembly of beta-cyclodextrin (β-CD)-modified generation 5 (G5) poly(amidoamine) (PAMAM) dendrimers as cores and adamantane (Ad)-decorated G3 PAMAM dendrimers as shells, and then partially modified the CSTD terminal amines with CBAA. The dendrimeric products were thoroughly characterized. Our results show that the obtained CSTDs display strong correlation signals between the Ad and β-CD protons in the two-dimensional NMR spectrum, indicating the strong host-guest interaction existing between G3-Ad and G5-CD in the structure of CSTDs. Diffusion NMR measurements reveal that the hydrodynamic diameter of CSTDs is smaller after partial CBAA surface modification, possibly due to the tightened molecular volume after zwitterionic modification. Lastly, the CBAA-modified CSTDs were demonstrated to have good antifouling property through protein resistance assay. Our findings provide a new strategy to study the framework of the developed CSTDs and CBAA-modified CSTDs, facilitating their further design and surface modification for various biomedical applications.

Benefits of time-resolved nonuniform sampling in reaction monitoring: The case of aza-Michael addition of benzylamine and acrylamide.

Monitoring of chemical reactions is best carried out using methods that sample the test object at a rate greater than the time scale of the processes taking place. The recently proposed time-resolved nonuniform sampling (TR-NUS) method allows the use of two-dimensional (2D) nuclear magnetic resonance (NMR) spectra for this purpose and provides a time resolution equivalent to that achievable using one-dimensional spectra. Herein, we show that TR-NUS acquired data eliminates 2D spectral line disturbances and enables more accurate signal integration and kinetics conclusions. The considerations are exemplified with a seemingly simple aza-Michael reaction of benzylamine and acrylamide. Surprisingly, the product identification is possible only using 2D spectra, although credible monitoring requires TR-NUS.

Isolation, purification, characterization and immunostimulatory activity of an exopolysaccharide produced by Lactobacillus pentosus LZ-R-17 isolated from Tibetan kefir

In this study, three strains of lactic acid bacteria isolated from Tibetan kefir grains, including two strains of Lactobacillus pentosus LZ-R-17 and L. helveticus LZ-R-5, and one strain of Lactococcus lactis subsp. lactis LZ-R-12. The ability of three strains to produce exopolysaccharide (EPS) was tested, and L. pentosus LZ-R-17 was found to have the highest EPS yield. One EPS (R-17-EPS) was isolated from the fermented milk by L. pentosus LZ-R-17 and purified by DEAE-52 anion exchange chromatography. Furthermore, R-17-EPS preliminary structure and macrophage immunomodulatory activity in vitro were investigated. On the basis of the analytical results of ultraviolet-visible spectrum, Fourier transform-infrared spectrum, monosaccharide composition analysis and one-dimensional and two-dimensional nuclear magnetic resonance (NMR) spectra, R-17-EPS was found to have an average molecular weight of 1.20 × 106 Da and was composed of galactose and glucose residues with a molar ratio of 1.00:3.15. NMR analysis revealed that the R-17-EPS was a linear hetero-galactoglucan containing repeating units of →2)-α-D-Galp-(1 → 4)-β-D-Glcp-(1 → 4)-β-D-Glcp-(1 → 4)-β-D-Glcp-(1→. In addition, R-17-EPS could effectively enhanced the proliferation, phagocytosis, nitric oxide and cytokines production of RAW264.7 cells, suggesting that R-17-EPS had potent immunostimulatory activity and could be explored as immunomodulator in functional food and/or medicine fields.

A new sesquiterpenoid from the shoots of Iranian Daphne mucronata Royle with selective inhibition of STAT3 and Smad3/4 cancer-related signaling pathways.

Daphne mucronata Royle grown in Iran has shown anticancer activities against different cancer cell lines. Therefore, within this study, we investigate the phytochemical pattern of this plant. Phytochemical investigation was done using standard column chromatography system: The structures were recognized by the interpretation of one and two-dimensional nuclear magnetic resonance (NMR) spectra and the help of High-Resolution Electrospray Ionization Mass spectroscopy (HR-ESIMS) and Infrared spectroscopy (IR) data. Stereochemistry was determined using 2D and 3D NOESY, and comparison of coupling constant values with literature. The absolute configuration was determined and confirmed using specific rotation and electronic circular dichroism experiments. Cytotoxicity was done against HeLa cells by standard MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Luciferase assay was used to check if the compounds can inhibit the activation of cancer-related signaling pathways. Molecular docking simulation was done for biological activity evaluation and to examine the interaction of the ligand with each of the proteins. A new sesquiterpenoid, 4,11(12)-guiadiene-1-ol-3-one (4), together with eight specialized metabolites, betulinic acid (1), coniferyl aldehyde (2), oleanolic acid (3), daphnetoxin (5), apigenin (7), syringin (8), and genkwanol A (9) were isolated and reported for the first time from the shoots of the plant. Compound 4 as an undescribed compound was submitted for cytotoxicity assay and showed moderate activity with the IC50 value of 51.3 ± 4.2μM against HeLa cancer cells. It showed selective inhibition of Interleukin-6 mediated signal transducer and activator of transcription 3 pathway (STAT-3/ IL-6), and Smad protein / transforming growth factor beta (TGF-β) transcription factors when screened through an array of cancer signaling pathways. Molecular docking confirmed biological tests and showed the interaction with STAT3 and Smad proteins. An undescribed sesquiterpenoid: 4,11(12)-guiadiene-1-ol-3-one in addition to eight known compounds were isolated. The new sesquiterpene was evaluated for the luciferase assay on 14 main cancer-related signaling pathways and showed selective inhibition of STAT3/IL6, and Smad/ TGF-β transcription factors. Molecular docking simulation showed more interactions with STAT3 than Smad, which confirms better interaction of compound 4 with STAT3 than Smad proteins. Graphical abstract.

Characterization of Reaction Products and Mechanisms between Serotonin and Methylglyoxal in Model Reactions and Mice.

Serotonin is an important endogenous regulatory neurotransmitter and has also been found in fruits, vegetables, and nuts. Methylglyoxal (MGO) is a reactive dicarbonyl metabolite and also a food toxin that modifies protein and DNA to cause the development of many chronic diseases. The objective of this study is to understand the reaction mechanisms between serotonin and MGO and determine whether serotonin could trap MGO in vivo. Five products were detected in phosphate buffer (pH 7.4) at 37 °C. Four products (compounds 2 and 4-6) were purified from the reaction mixture, and their structures were characterized by the analysis of their high-resolution mass and one- and two-dimensional nuclear magnetic resonance spectra. One product (compound 3), as a result of its instability, could not be properly purified and was tentatively characterized on the basis of its high-resolution mass spectrum and corresponding mass fragments. On the basis of the structures of these five products, two reaction pathways were proposed. Compounds 2, 3, 5, and 6 were produced through the Pictet-Spengler condensation pathway between the primary amine of serotonin and the ketone of MGO, and compound 3 was identified as the intermediate product to form products 2, 5, and 6, whereas compound 4 was formed through nucleophilic substitution by the benzene ring of serotonin, which is a new reaction pathway between biogenic amines and reactive carbonyl species. More importantly, the detection of adducts 2 and 4-6 in mice supports our hypothesis that the reaction between serotonin and MGO also happens in vivo through the same pathways as those in model reactions, suggesting that dietary or endogenous serotonin has the capacity to trap MGO in vivo.

Low-field benchtop NMR spectroscopy: status and prospects in natural product analysis†.

Since a couple of years, low-field (LF) nuclear magnetic resonance (NMR) spectrometers (40-100 MHz) have re-entered the market. They are used for various purposes including analyses of natural products. Similar to high-field instruments (300-1200 MHz), modern LF instruments can measure multiple nuclei and record two-dimensional (2D) NMR spectra. To review the commercial availability as well as applications, advantages, limitations, and prospects of LF-NMR spectrometers for the purpose of natural products analysis. Commercial LF instruments were compared. A literature search was performed for articles using and discussing modern LF-NMR. Next, the articles relevant to natural products were read and summarised. Seventy articles were reviewed. Most appeared in 2018 and 2019. Low costs and ease of operation are most often mentioned as reasons for using LF-NMR. As the spectral resolution of LF instruments is limited, they are not used for structure elucidation of new natural products but rather applied for quality control (QC), forensics, food and health research, process control and teaching. Chemometric data handling is valuable. LF-NMR is a rapidly developing niche and new instruments keep being introduced.

Everything you wanted to know about phase and reference frequency in one- and two-dimensional NMR spectroscopy.

The fundamental concept of phase discussed in this tutorial aimed at providing students with an explanation of the delays and processing parameters they may find in nuclear magnetic resonance (NMR) pulse programs. We consider the phase of radio-frequency pulses, receiver, and magnetization and how all these parameters are related to phases and offsets of signals in spectra. The impact of the off-resonance effect on the phase of the magnetization is discussed before presenting an overview of how adjustment of the time reference of the free induction decay avoids first-order correction of the phase of spectra. The main objective of this tutorial is to show how the relative phase of a pulse and the receiver can be used to change the reference frequency along direct and indirect dimensions of NMR experiments. Unusual of phase incrementation with non-90° angles will be illustrated on one- and two-dimensional NMR spectra.

Identification of Light Oil in 2D NMR Spectra of Tight Sandstone Reservoirs by Using L1/L2 Two-Parameter Regularization

This work presents L1/L2 two-parameter regularization as an efficient technique for the identification of light oil in the two-dimensional (2D) nuclear magnetic resonance (NMR) spectra of tight san...

Unusual stilbene glucosides from Vitis vinifera roots

Aim: Stilbenes are well-known phytoalexins present in vine and wine. Stilbene glucosides have been identified in wine; however, other than piceid, these compounds have never been reported to be present in the woody parts of grapevine. The aims of this study were to investigate the presence of stilbene glucosides in the woody parts of the vine and to evaluate their cytotoxic activity, in comparison with that of resveratrol, in human hepatoma HepG2 cells.Methods and results: Stilbene glucosides were isolated from a Vitis vinifera root extract. The extract was partitioned with ethyl acetate and fractionated by polyamide gel column chromatography. Pure compounds were obtained by semipreparative high-performance liquid chromatography. These were then identified by mass spectrometry and nuclear magnetic resonance (NMR) analyses, including analysis of two-dimensional NMR spectra. In addition to resveratrol, five stilbene glucosides were found: resveratroloside, resveratrol rutinoside, trans-ε-viniferin diglucoside, cis-ε-viniferin diglucoside and piceid. Of these, the first four showed cytotoxic effects against HepG2 cells when the crystal violet assay was used to determine cell viability.Conclusion: In addition to resveratrol and piceid, this is the first report of the presence of four glucosylated derivatives of resveratrol (resveratroloside, resveratrol rutinoside, and trans- and cis-ε-viniferin diglucosides) in the woody parts of vine. These compounds showed significant cytotoxicity against HepG2 cells.Significance and impact of the study: Stilbenes are well-known biological compounds. Grapevine is one of the main sources of this family of polyphenols. Other than piceid, stilbene glucosides have been identified in wine but never in the woody parts of vine. This is the first study in which five glucosylated derivatives of resveratrol were isolated from woody parts of vine. They were also shown to exert antiproliferative effects in human hepatoma HepG2 cells.