Enhanced NMR Spectroscopy Research Articles

Unravelling the Molecular Structure and Confining Environment of an Organometallic Catalyst Heterogenized within Amorphous Porous Polymers.

The catalytic activity of multifunctional, microporous materials is directly linked to the spatial arrangement of their structural building blocks. Despite great achievements in the design and incorporation of isolated catalytically active metal complexes within such materials, a detailed understanding of their atomic-level structure and the local environment of the active species remains a fundamental challenge, especially when these latter are hosted in non-crystalline organic polymers. Here, we show that by combining computational chemistry with pair distribution function analysis, 129 Xe NMR, and Dynamic Nuclear Polarization enhanced NMR spectroscopy, a very accurate description of the molecular structure and confining surroundings of a catalytically active Rh-based organometallic complex incorporated inside the cavity of amorphous bipyridine-based porous polymers is obtained. Small, but significant, differences in the structural properties of the polymers are highlighted depending on their backbone motifs.

Aufklärung der molekularen Struktur und des Confinements eines in amorphen porösen Polymeren heterogenisierten metallorganischen Katalysators**

AbstractDie katalytische Aktivität multifunktionaler, mikroporöser Materialien hängt direkt von der räumlichen Anordnung ihrer Strukturbausteine ab. Trotz großer Erfolge bei dem Design und der Heterogenisierung isolierter, katalytisch aktiver Metallkomplexe in solchen Materialien bleibt ein detailliertes Verständnis ihrer Struktur auf atomarer Ebene und der lokalen Umgebung der aktiven Spezies eine grundlegende Herausforderung, insbesondere wenn diese in amorphe organische Polymere eingebaut sind. Hier zeigen wir, dass durch die Kombination von Computerchemie mit Paarverteilungsfunktionsanalyse, 129Xe NMR‐ und dynamischer Kernpolarisations NMR‐Spektroskopie eine sehr genaue Beschreibung der Molekülstruktur und der Umgebung eines katalytisch aktiven, Rh‐basierten organometallischen Komplexes möglich ist, der in die Poren eines amorphen Bipyridin‐basierten porösen Polymers eingebaut ist. Kleine, aber signifikante Unterschiede in den strukturellen Eigenschaften der Polymere in Abhängigkeit ihrer Funktionalisierung werden hervorgehoben.

INTERFACES. A program for determining the 3D structures of surfaces sites using NMR data

Dynamic nuclear polarization surface enhanced NMR spectroscopy has enabled the determination of high-resolution structures from surface-supported molecules, including single-site heterogeneous catalysts. Structure determinations have largely mimicked the approaches used in biomolecular NMR spectroscopy, namely, using distance measurements to constrain a conformational search. These early demonstrations made use of purpose-built software, which has limited the adoption of the technique. Herein, we describe the open-source program INTERFACES (Interpret NMR to Elucidate or Reconstruct the Full Atomistic Configurations of External Surfaces) which automates the analysis of RE(SP)DOR data as well as the structure determination for surface sites. Distances, angles, dihedral angles, complex orientation, and distance from the support can all be sampled to find all structures that agree with the experimental data. A χ2 metric is used to define the error ranges of the REDOR fits and produce structures with an arbitrary level of confidence. Structural solutions are then provided as both overlays and ORTEP-like probability ellipsoids.

CW-EPR spectra of P1 centers in HPHT diamond in the vicinity of Rabi resonance: possible standard for [formula omitted] evaluation in EPR and DNP enhanced NMR spectroscopies

Synthetic commercial type-IIa diamond crystal containing low concentration of P1 (Ns0) centers ([Ns0] <1ppm) was examined as a possible calibration standard for evaluating microwave magnetic field intensity (B1) in the spectrometer cavity. The reliability of this standard was experimentally and theoretically tested by employing two different types of microwave cavities with different distributions of B1 values. The procedure relies on the ordinary continuous wave electron paramagnetic resonance (CW-EPR) with magnetic field modulation (ωrf/2π=100 kHz) applied under the regime of the microwave power (PMW) saturation of the out-of-phase first harmonic signal. The power saturation procedure for PMW∼B11/2 was analyzed with respect to the change of spectral lineshape when Rabi frequency (ω1=γB1, γis the electron gyromagnetic ratio) approaches to ωrf. The phenomenon of the inversion of sideband lines while passing the Rabi resonance condition (ω1=ωrf) was analyzed. In specific, from the simulation of the experimental lineshapes in the close vicinity of Rabi resonance (ω1≈ωrf) the correspondingB1 can be assigned. For the chosen diamond crystal low value of B1≈3.6µT at Rabi resonance was determined what makes it a convenient standard for DNP enhanced NMR spectroscopy which requires relatively small B1.

Contrasting Photochemical and Thermal Catalysis by Ruthenium Arsine Complexes Revealed by Parahydrogen Enhanced NMR Spectroscopy

AbstractThe thermal and photochemical reactivity of Ru(CO)3(dpae) (1) and Ru(CO)2(dpae)(PPh3) (2) towards H2 and diphenylacetylene is described. These reactions are monitored by NMR spectroscopy in conjunction with the parahydrogen induced polarisation (PHIP) effect, spatially resolved chemical shift imaging and the Only Parahydrogen Spectroscopy (OPSY) signal filtering method. The results are supported by DFT. The thermal and photochemical reactions of 1 with H2 proceed by CO loss and form Ru(H)2(CO)2(dpae) (3). 1 catalyses the formation of 1,2 diphenylethane, cis‐ and trans‐stilbene, and 1,2,3,4 tetraphenylbutadiene under 325 nm irradiation at 295 K in a reaction where Ru(CO)2(dpae)(η2‐CHPh=CPhCPh=CHPh) forms. When the same reaction is monitored under thermal conditions at 333 K the η2‐diene complex is no longer detected but hydride containing Ru(CHPhCH2Ph)(H)(CO)2(dpae) and Ru(CO)2(dpae)(trans‐stilbene) are seen. For 1, the photochemical promotion of hydrogenation through 325 nm irradiation results in an approximate 5.5‐fold increase in turnover at 333 K when compared to no irradiation. In contrast, 2 reacts thermally with H2 at 295 K through PPh3 and CO loss with both Ru(H)2(CO)(dpae)(solvent) and 3 being detected. Under irradiation, CO loss dominates and two isomers of Ru(H)2(CO)(dpae)(PPh3) form. While 2 forms the same 4 organic products at 295 K and a second isomer of Ru(CHPhCH2Ph)(H)(CO)2(dpae) alongside the diene complex no photochemical promotion of hydrogenation is observed.

Structural Characterization of Phosphate Species Adsorbed on γ-Alumina by Combining DNP Surface Enhanced NMR Spectroscopy and DFT Calculations

Structural Characterization of Phosphate Species Adsorbed on γ-Alumina by Combining DNP Surface Enhanced NMR Spectroscopy and DFT Calculations

Monitoring Crystallization Processes in Confined Porous Materials by Dynamic Nuclear Polarization Solid-State Nuclear Magnetic Resonance.

Establishing mechanistic understanding of crystallization processes at the molecular level is challenging, as it requires both the detection of transient solid phases and monitoring the evolution of both liquid and solid phases as a function of time. Here, we demonstrate the application of dynamic nuclear polarization (DNP) enhanced NMR spectroscopy to study crystallization under nanoscopic confinement, revealing a viable approach to interrogate different stages of crystallization processes. We focus on crystallization of glycine within the nanometric pores (7–8 nm) of a tailored mesoporous SBA-15 silica material with wall-embedded TEMPO radicals. The results show that the early stages of crystallization, characterized by the transition from the solution phase to the first crystalline phase, are straightforwardly observed using this experimental strategy. Importantly, the NMR sensitivity enhancement provided by DNP allows the detection of intermediate phases that would not be observable using standard solid-state NMR experiments. Our results also show that the metastable β polymorph of glycine, which has only transient existence under bulk crystallization conditions, remains trapped within the pores of the mesoporous SBA-15 silica material for more than 200 days.

Direct Observation of Carbonate Formation in Partly Hydrated Tricalcium Silicate by Dynamic Nuclear Polarization Enhanced NMR Spectroscopy

Solid-state NMR combined with dynamic nuclear polarization (DNP NMR) is used to study hydration processes in tricalcium silicate (Ca3SiO5, abbreviated as C3S) samples. The studied C3S samples have experienced early stage hydration (1–30 h) and slow aging (9 years) processes. The appearance of Q3 and Q4 lines in the 29Si MAS and 1H → 29Si CP MAS NMR spectra obtained for partly hydrated C3S samples indicated the formation of amorphous silica which corresponds to their carbonation, which was corroborated by complementary FTIR data. Significant DNP signal enhancements obtained for the studied samples allowed to further investigate the C3S carbonation process in detail employing the 1H → 29Si CP MAS FSLG HETCOR technique. Finally, DNP enhanced 1H → 13C CP MAS and 1H → 13C CP MAS FSLG HETCOR techniques enabled to directly observe the formation of carbonate moieties in partly hydrated C3S samples.

Uncovering selective and active Ga surface sites in gallia-alumina mixed-oxide propane dehydrogenation catalysts by dynamic nuclear polarization surface enhanced NMR spectroscopy.

Gallia–alumina (Ga,Al)2O3(x : y) spinel-type solid solution nanoparticle catalysts for propane dehydrogenation (PDH) were prepared with four nominal Ga : Al atomic ratios (1 : 6, 1 : 3, 3 : 1, 1 : 0) using a colloidal synthesis approach. The structure, coordination environment and distribution of Ga and Al sites in these materials were investigated by X-ray diffraction, X-ray absorption spectroscopy (Ga K-edge) as well as 27Al and 71Ga solid state nuclear magnetic resonance. The surface acidity (Lewis or Brønsted) was probed using infrared spectroscopy with pyridine and 2,6-dimethylpyridine probe molecules, complemented by element-specific insights (Ga or Al) from dynamic nuclear polarization surface enhanced cross-polarization magic angle spinning 15N{27Al} and 15N{71Ga} J coupling mediated heteronuclear multiple quantum correlation NMR experiments using 15N-labelled pyridine as a probe molecule. The latter approach provides unique insights into the nature and relative strength of the surface acid sites as it allows to distinguish contributions from Al and Ga sites to the overall surface acidity of mixed (Ga,Al)2O3 oxides. Notably, we demonstrate that (Ga,Al)2O3 catalysts with a high Al content show a greater relative abundance of four-coordinated Ga sites and a greater relative fraction of weak/medium Ga-based surface Lewis acid sites, which correlates with superior propene selectivity, Ga-based activity, and stability in PDH (due to lower coking). In contrast, (Ga,Al)2O3 catalysts with a lower Al content feature a higher fraction of six-coordinated Ga sites, as well as more abundant Ga-based strong surface Lewis acid sites, which deactivate through coking. Overall, the results show that the relative abundance and strength of Ga-based surface Lewis acid sites can be tuned by optimizing the bulk Ga : Al atomic ratio, thus providing an effective measure for a rational control of the catalyst performance.

Ultra-Low Loading Pt/CeO2 Catalysts: Ceria Facet Effect Affords Improved Pairwise Selectivity for Parahydrogen Enhanced NMR Spectroscopy.

Oxide supports with well-defined shapes enable investigations on the effects of surface structure on metal-support interactions and correlations to catalytic activity and selectivity. Here, a modified atomic layer deposition technique was developed to achieve ultra-low loadings (8-16 ppm) of Pt on shaped ceria nanocrystals. Using octahedra and cubes, which expose exclusively (111) and (100) surfaces, respectively, the effect of CeO2 surface facet on Pt-CeO2 interactions under reducing conditions was revealed. Strong electronic interactions result in electron-deficient Pt species on CeO2 (111) after reduction, which increased the stability of the atomically dispersed Pt. This afforded significantly higher NMR signal enhancement in parahydrogen-induced polarization experiments compared with the electron-rich platinum on CeO2 (100), and a factor of two higher pairwise selectivity (6.1 %) in the hydrogenation of propene than any previously reported monometallic heterogeneous Pt catalyst.

Ultra‐Low Loading Pt/CeO2 Catalysts: Ceria Facet Effect Affords Improved Pairwise Selectivity for Parahydrogen Enhanced NMR Spectroscopy

AbstractOxide supports with well‐defined shapes enable investigations on the effects of surface structure on metal–support interactions and correlations to catalytic activity and selectivity. Here, a modified atomic layer deposition technique was developed to achieve ultra‐low loadings (8–16 ppm) of Pt on shaped ceria nanocrystals. Using octahedra and cubes, which expose exclusively (111) and (100) surfaces, respectively, the effect of CeO2 surface facet on Pt‐CeO2 interactions under reducing conditions was revealed. Strong electronic interactions result in electron‐deficient Pt species on CeO2 (111) after reduction, which increased the stability of the atomically dispersed Pt. This afforded significantly higher NMR signal enhancement in parahydrogen‐induced polarization experiments compared with the electron‐rich platinum on CeO2 (100), and a factor of two higher pairwise selectivity (6.1 %) in the hydrogenation of propene than any previously reported monometallic heterogeneous Pt catalyst.

The Structure of Molecular and Surface Platinum Sites Determined by DNP-SENS and Fast MAS 195Pt Solid-State NMR Spectroscopy.

The molecular level characterization of heterogeneous catalysts is challenging due to the low concentration of surface sites and the lack of techniques that can selectively probe the surface of a heterogeneous material. Here, we report the joint application of room temperature proton-detected NMR spectroscopy under fast magic angle spinning (MAS) and dynamic nuclear polarization surface enhanced NMR spectroscopy (DNP-SENS), to obtain the 195Pt solid-state NMR spectra of a prototypical example of highly dispersed Pt sites (single site or single atom), here prepared via surface organometallic chemistry, by grafting [(COD)Pt(OSi(OtBu)3)2] (1, COD = 1,5-cyclooctadiene) on partially dehydroxylated silica (1@SiO2). Compound 1@SiO2 has a Pt loading of 3.7 wt %, a surface area of 200 m2/g, and a surface Pt density of around 0.6 Pt site/nm2. Fast MAS 1H{195Pt} dipolar-HMQC and S-REDOR experiments were implemented on both the molecular precursor 1 and on the surface complex 1@SiO2, providing access to 195Pt isotropic shifts and Pt-H distances, respectively. For 1@SiO2, the measured isotropic shift and width of the shift distribution constrain fits of the static wide-line DNP-enhanced 195Pt spectrum, allowing the 195Pt chemical shift tensor parameters to be determined. Overall the NMR data provide evidence for a well-defined, single-site structure of the isolated Pt sites.

Atomic-Scale Description of Interfaces between Antigen and Aluminum-Based Adjuvants Used in Vaccines by Dynamic Nuclear Polarization (DNP) Enhanced NMR Spectroscopy.

The addition of aluminum-based adjuvants in vaccines enhances the immune response to antigens. The strength of antigen adsorption on adjuvant gels is known to modulate vaccine efficacy. However, a detailed understanding of the mechanisms of interaction between aluminum gels and antigens is still missing. Herein, a new analytical approach based on dynamic nuclear polarization (DNP) enhanced NMR spectroscopy under magic angle spinning (MAS) is implemented to provide a molecular description of the antigen-adjuvant interface. This approach is demonstrated on hepatitis B surface antigen particles in combination with three aluminum gels obtained from different suppliers. Both noncovalent and covalent interactions between the phospholipids of the antigen particles and the surface of the aluminum gels are identified by using MAS DNP NMR 27 Al and 31 P correlation experiments. Although covalent interactions were detected for only one of the formulations, dipolar recoupling rotational echo adiabatic passage double resonance (REAPDOR) experiments reveal significant differences in the strength of weak interactions.

Probing Functionalities and Acidity of Calcined Phenylene-Bridged Periodic Mesoporous Organosilicates Using Dynamic Nuclear Polarization NMR, Diffuse Reflectance Infrared Fourier Transform Spectroscopy, and X-ray Photoelectron Spectroscopy

Owing to their high surface area, their high stability, and their hydrophobicity, periodic mesoporous organosilica (PMO) materials represent promising catalytic support for environmentally friendly chemical processes in water. We investigate here how the calcination of PMO material with benzene linkers (PMOB) allows its functionalization. Conventional and dynamic nuclear polarization (DNP)-enhanced NMR spectroscopy, diffuse reflectance infrared Fourier transform spectroscopy, and X-ray photoelectron spectroscopy prove that calcination at 450 {\deg}C results in the oxidation of phenylene bridges into (poly)phenols but also into carboxylic acids. Ketone, aldehyde, as well as allyl and aliphatic alcohol functionalities are also observed, but their amount is much lower than that of carboxylic acids. The calcination also cleaves the Si-C bonds. Nevertheless, N2 adsorption desorption measurements, powder X-ray diffraction, and transmission electron microscopy indicate that the PMOB materials calcined up to 600 {\deg}C still exhibit ordered mesopores. We show that the phenol and carboxylic acid functionalities of PMOB calcined at 450 {\deg}C protonate the NH2 group of 1-(3-aminopropyl)imidazole (API) in water at room temperature, but no formation of a covalent bond between API and the calcined PMOB functionalities has been detected.

TinyPols: a family of water-soluble binitroxides tailored for dynamic nuclear polarization enhanced NMR spectroscopy at 18.8 and 21.1 T.

Dynamic Nuclear Polarization (DNP) has recently emerged as a key method to increase the sensitivity of solid-state NMR spectroscopy under Magic Angle Spinning (MAS). While efficient binitroxide polarizing agents such as AMUPol have been developed for MAS DNP NMR at magnetic fields up to 9.4 T, their performance drops rapidly at higher fields due to the unfavorable field dependence of the cross-effect (CE) mechanism and AMUPol-like radicals were so far disregarded in the context of the development of polarizing agents for very high-field DNP. Here, we introduce a new family of water-soluble binitroxides, dubbed TinyPols, which have a three-bond non-conjugated flexible amine linker allowing sizable couplings between the two unpaired electrons. We show that this adjustment of the linker is crucial and leads to unexpectedly high DNP enhancement factors at 18.8 T and 21.1 T: an improvement of about a factor 2 compared to AMUPol is reported for spinning frequencies ranging from 5 to 40 kHz, with εH of up to 90 at 18.8 T and 38 at 21.1 T for the best radical in this series, which are the highest MAS DNP enhancements measured so far in aqueous solutions at these magnetic fields. This work not only breathes a new momentum into the design of binitroxides tailored towards high magnetic fields, but also is expected to push the application frontiers of high-resolution DNP MAS NMR, as demonstrated here on a hybrid mesostructured silica material.

Imine Metathesis by Silica-Supported Catalysts Using the Methodology of Surface Organometallic Chemistry.

With this protocol, a well-defined singlesite silica-supported heterogeneous catalyst [(≡Si-O-)Hf(=NMe)(η1-NMe2)] is designed and prepared according to the methodology developed by surface organometallic chemistry (SOMC). In this framework, catalytic cycles can be determined by isolating crucial intermediates. All air-sensitive materials are handled under inert atmosphere (using gloveboxes or a Schlenk line) or high vacuum lines (HVLs, <10-5 mbar). The preparation of SiO2-700 (silica dehydroxylated at 700 °C) and subsequent applications (the grafting of complexes and catalytic runs) requires the use of HVLs and double-Schlenk techniques. Several well-known characterization methods are used, such as Fourier-transform infrared spectroscopy (FTIR), elemental microanalysis, solid-state nuclear magnetic resonance spectroscopy (SSNMR), and state-of-the-art dynamic nuclear polarization surface enhanced NMR spectroscopy (DNP-SENS). FTIR and elemental microanalysis permit scientists to establish the grafting and its stoichiometry. 1H and 13C SSNMR allows the structural determination of the hydrocarbon ligands coordination sphere. DNP SENS is an emerging powerful technique in solid characterization for the detection of poorly sensitive nuclei (15N, in our case). SiO2-700 is treated with about one equivalent of the metal precursor compared to the amount of surface silanol (0.30 mmol·g-1) in pentane at room temperature. Then, volatiles are removed, and the powder samples are dried under dynamic high vacuum to afford the desired materials [(≡Si-O-)Hf(η2π-MeNCH2)(η1-NMe2)(η1-HNMe2)]. After a thermal treatment under high vacuum, the grafted complex is converted into metal imido silica complex [(≡Si-O-)Hf(=NMe)(η1-NMe2)]. [(≡Si-O-)Hf(=NMe)(η1-NMe2)] effectively promotes the metathesis of imines, using the combination of two imine substrates, N-(4-phenylbenzylidene)benzylamine, or N-(4-fluorobenzylidene)-4-fluoroaniline, with N-benzylidenetert-butylamine as substrates. A significantly lower conversion is observed with the blank runs; thus, the presence of the imido group in [(≡Si-O-)Hf(=NMe)(η1-NMe2)] is correlated to the catalytic performance.

Efficient Building Blocks for Solid-Phase Peptide Synthesis of Spin Labeled Peptides for Electron Paramagnetic Resonance and Dynamic Nuclear Polarization Applications.

Specific spin labeling allows the site-selective investigation of biomolecules by EPR and DNP enhanced NMR spectroscopy. A novel spin labeling strategy for commercially available Fmoc-amino acids is developed. In this approach, the PROXYL spin label is covalently attached to the hydroxyl side chain of three amino acids hydroxyproline (Hyp), serine (Ser) and tyrosine (Tyr) by a simple three-step synthesis route. The obtained PROXYL containing building-blocks are N-terminally protected by the Fmoc-protection group, which makes them applicable for the use in solid-phase peptide synthesis (SPPS). This approach allows the insertion of the spin label at any desired position during SPPS, which makes it more versatile than the widely used post synthetic spin labeling strategies. For the final building-blocks, the radical activity is proven by EPR. DNP enhanced solid-state NMR experiments employing these building-blocks in a TCE solution show enhancement factors of up to 26 for 1 H and 13 C (1 H→13 C cross-polarization). To proof the viability of the presented building-blocks for insertion of the spin label during SPPS the penta-peptide Acetyl-Gly-Ser(PROXYL)-Gly-Gly-Gly was synthesized employing the spin labeled Ser building-block. This peptide could successfully be isolated and the spin label activity proved by EPR and DNP NMR measurements, showing enhancement factors of 12.1±0.1 for 1 H and 13.9±0.5 for 13 C (direct polarization).

19F Magic Angle Spinning Dynamic Nuclear Polarization Enhanced NMR Spectroscopy

AbstractThe introduction of high‐frequency, high‐power microwave sources, tailored biradicals, and low‐temperature magic angle spinning (MAS) probes has led to a rapid development of hyperpolarization strategies for solids and frozen solutions, leading to large gains in NMR sensitivity. Here, we introduce a protocol for efficient hyperpolarization of 19F nuclei in MAS DNP enhanced NMR spectroscopy. We identified trifluoroethanol‐d3 as a versatile glassy matrix and show that 12 mm AMUPol (with microcrystalline KBr) provides direct 19F DNP enhancements of over 100 at 9.4 T. We applied this protocol to obtain DNP‐enhanced 19F and 19F–13C cross‐polarization (CP) spectra for an active pharmaceutical ingredient and a fluorinated mesostructured hybrid material, using incipient wetness impregnation, with enhancements of approximately 25 and 10 in the bulk solid, respectively. This strategy is a general and straightforward method for obtaining enhanced 19F MAS spectra from fluorinated materials.

19 F Magic Angle Spinning Dynamic Nuclear Polarization Enhanced NMR Spectroscopy.

The introduction of high-frequency, high-power microwave sources, tailored biradicals, and low-temperature magic angle spinning (MAS) probes has led to a rapid development of hyperpolarization strategies for solids and frozen solutions, leading to large gains in NMR sensitivity. Here, we introduce a protocol for efficient hyperpolarization of 19 F nuclei in MAS DNP enhanced NMR spectroscopy. We identified trifluoroethanol-d3 as a versatile glassy matrix and show that 12 mm AMUPol (with microcrystalline KBr) provides direct 19 F DNP enhancements of over 100 at 9.4 T. We applied this protocol to obtain DNP-enhanced 19 F and 19 F-13 C cross-polarization (CP) spectra for an active pharmaceutical ingredient and a fluorinated mesostructured hybrid material, using incipient wetness impregnation, with enhancements of approximately 25 and 10 in the bulk solid, respectively. This strategy is a general and straightforward method for obtaining enhanced 19 F MAS spectra from fluorinated materials.

Identification of metabolites in coriander seeds (Coriandrum Sativum L.) aided by ultrahigh resolution total correlation NMR spectroscopy.

NMR is a fast method for obtaining a holistic snapshot of the metabolome and also offers quantitative information without separating the compounds present in a complex mixture. Identification of the metabolites present in a plant extract sample is a crucial step for all plant metabolomics studies. In the present work, we used various two dimensional (2D) NMR methods such as J-resolved NMR, total correlation spectroscopy (TOCSY), and heteronuclear single quantum coherence sensitivity enhanced NMR spectroscopy for the identification of 36 common metabolites present in Coriandrum sativum L. seed extract. The identified metabolites belong to the following classes: organic acids, amino acids, and carbohydrates. 1 H NMR spectra of such complex mixtures in general display tremendous signal overlap due to the presence of a large number of metabolites with closely resonating multiplet signals. This signal overlapping leads to ambiguity in an assignment, and hence, identification of metabolites becomes tedious or impossible in many cases. Therefore, the utility of pure-shift proton spectrum along the indirect (F1 ) dimension of the F1 -PSYCHE-TOCSY spectrum is demonstrated for overcoming ambiguity in assignment of metabolites in crowded spectral regions from Coriandrum sativum L. seed extract sample. Because pure-shift NMR methods yield ultrahigh resolution spectrum (i.e., a singlet peak per chemical site) along one or more dimensions, such spectra provide better identification of metabolites compared with regular 2D TOCSY where signal overlap and peak distortions lead to ambiguity in the assignment. Nine metabolites were unambiguously assigned by pure-shift F1 -PSYCHE-TOCSY spectrum, which was unresolved in regular TOCSY spectrum.