OH Proton Research Articles

Solution to doubling spectroscopy challenge.

The reason for the signal doubling of the H-6 hydrogen signal in the H-NMR spectrum of dexelvucitabine (DFC) is the presence of the fluorine at position 5 (Fig. 1). Fluorine has only one isotope, F, with a spin 1⁄2 nucleus. Since both H-6 and F-5 are connected in DFC by a vicinal coupling JH6,F5, the NMR signal of each H-6 nucleus is split by the interaction with the F-5 nucleus and vice versa. Therefore, no cross-peak will be observed for the H-6 doublet in the H,H-COSY spectrum. As expected, noticeable crosspeaks were observed in the NOESY spectrum for interactions between H-6, H-2′, H-5′, and –OH protons, indicating correct signal assignment (Fig. 2). Considerable cross-peaks were also observed between the pair of twin amino proton signals in the NOESY spectrum. However, the phase of these cross-peaks (red color) is opposite to that of the other NOESY cross-peaks (blue color). This indicates that there is a slow exchange process between both amino protons during the 700-ms continuous mixing time at room temperature. The energy barrier for this exchange process can be calculated using the experimentally determined coalescence temperature Tc and the frequency difference Δν between the separate signals of the amino-group protons. Both of these parameters were determined twice at different magnetic field strengths:

Analysis of Proton NMR in Hydrogen Bonds in Terms of Lone-Pair and Bond Orbital Contributions.

NMR spectroscopic parameters of the proton involved in hydrogen bonding are studied theoretically. The set of molecules includes systems with internal resonance-assisted hydrogen bonds, internal hydrogen bonds but no resonance stabilization, the acetic acid dimer (AAD), a DNA base pair, and the hydrogen succinate anion (HSA). Ethanol and guanine represent reference molecules without hydrogen bonding. The calculations are based on zero-point vibrationally averaged molecular structures in order to include anharmonicity effects in the NMR parameters. An analysis of the calculated NMR shielding and J-coupling is performed in terms of "chemist's orbitals", that is, localized molecular orbitals (LMOs) representing lone-pairs, atomic cores, and bonds. The LMO analysis associates some of the strong de-shielding of the protons in resonance-assisted hydrogen bonds with delocalization involving the π-backbone. Resonance is also shown to be an important factor causing de-shielding of the OH protons for AAD and HSA, but not for the DNA base pair. Nitromalonamide (NMA) and HSA have particularly strong hydrogen bonds exhibiting signs of covalency in the associated J-couplings. The analysis results show how NMR spectroscopic parameters that are characteristic for hydrogen bonded protons are influenced by the geometry and degree of covalency of the hydrogen bond as well as intra- and intermolecular resonance.

(1)H NMR spectra of alcohols in hydrogen bonding solvents: DFT/GIAO calculations of chemical shifts.

Proton nuclear magnetic resonance (NMR) shifts of aliphatic alcohols in hydrogen bonding solvents have been computed on the basis of density functional theory by applying the gauge-including atomic orbital method to geometry-optimized alcohol/solvent complexes. The OH proton shifts and hydrogen bond distances for methanol or ethanol complexed with pyridine depend very much on the functional employed and very little on the basis set, provided it is sufficiently large to give the correct quasi-linear hydrogen bond geometry. The CH proton shifts are insensitive to both the functional and the basis set. NMR shifts for all protons in several alcohol/pyridine complexes are calculated at the Perdew, Burke and Ernzerhof PBE0/cc-pVTZ//PBE0/6-311 + G(d,p) level in the gas phase. The results correlate with the shifts for the pyridine-complexed alcohols, determined by analysing data from the NMR titration of alcohols against pyridine. More pragmatically, computed shifts for a wider range of alcohols correlate with experimental shifts in neat pyridine. Shifts for alcohols in dimethylsulfoxide, based on the corresponding complexes in the gas phase, correlate well with the experimental values, but the overall root mean square difference is high (0.23 ppm), shifts for the OH, CHOH and other CH protons being systematically overestimated, by averages of 0.42, 0.21 and 0.06 ppm, respectively. If the computed shifts are corrected accordingly, a very good correlation is obtained with a gradient of 1.00 ± 0.01, an intercept of 0.00 ± 0.02 ppm and a root mean square difference of 0.09 ppm. This is a modest improvement on the result of applying the CHARGE programme to a slightly different set of alcohols. Some alcohol complexes with acetone and acetonitrile were investigated both in the gas phase and in a continuum of the relevant solvent.

Nuclear spin hyperpolarization of the solvent using signal amplification by reversible exchange (SABRE)

Here we report the polarization of the solvent OH protons by SABRE using standard iridium-based catalysts under slightly acidic conditions. Solvent polarization was observed in the presence of a variety of structurally similar N-donor substrates while no solvent enhancement was observed in the absence of substrate or para-hydrogen (p-H2). Solvent polarization was sensitive to the polarizing field and catalyst:substrate ratio in a manner similar to that of substrate protons. SABRE experiments with pyridine-d5 suggest a mechanism where hyperpolarization is transferred from the free substrate to the solvent by chemical exchange while measured hyperpolarization decay times suggest a complimentary mechanism which occurs by direct coordination of the solvent to the catalytic complex. We found the solvent hyperpolarization to decay nearly 3 times more slowly than its characteristic spin–lattice relaxation time suggesting that the hyperpolarized state of the solvent may be sufficiently long lived (∼20s) to hyperpolarize biomolecules having exchangeable protons. This route may offer future opportunities for SABRE to impact metabolic imaging.

Label-free in vivo molecular imaging of underglycosylated mucin-1 expression in tumour cells.

Alterations in mucin expression and glycosylation are associated with cancer development. Underglycosylated mucin-1 (uMUC1) is overexpressed in most malignant adenocarcinomas of epithelial origin (for example, colon, breast and ovarian cancer). Its counterpart MUC1 is a large polymer rich in glycans containing multiple exchangeable OH protons, which is readily detectable by chemical exchange saturation transfer (CEST) MRI. We show here that deglycosylation of MUC1 results in >75% reduction in CEST signal. Three uMUC1+ human malignant cancer cell lines overexpressing uMUC1 (BT20, HT29 and LS174T) show a significantly lower CEST signal compared with the benign human epithelial cell line MCF10A and the uMUC1− tumour cell line U87. Furthermore, we demonstrate that in vivo CEST MRI is able to make a distinction between LS174T and U87 tumour cells implanted in the mouse brain. These results suggest that the mucCEST MRI signal can be used as a label-free surrogate marker to non-invasively assess mucin glycosylation and tumour malignancy.

Catalysis effect of micro-hydration on the intramolecular proton transfer in cytosine

Structural and thermodynamic properties of nine isomers of cytosine were studied in gas and aqueous phases and in micro-hydrated environment employing B3LYP and MP2 methods. Also, isomerizations of cytosine were studied in gas phase and the activation energies (Ea) and Gibbs free energies (ΔG#) of the internal OH rotations and proton transfer processes were calculated. The calculated OH rotational barriers were smaller than 50kJ/mol while the activation energies of intramolecular proton transfers were in the range of 110–190kJ/mol. Effect of mono- and di-hydration of the cytosine isomers on the transition state structures and the energy barriers was investigated.

Antioxidant properties of xanthones extracted from the pericarp of Garcinia mangostana (Mangosteen): A theoretical study

A theoretical study on antioxidant properties of fourteen xanthones extracted from the pericarp of G. Mangostana has been performed. Three main reaction mechanisms are investigated: hydrogen atom transfer (HAT), single electron transfer–proton transfer (SETPT) and sequential proton loss electron transfer (SPLET). The OH bond dissociation enthalpy (BDE), ionization energy (IE), proton affinity (PA) and electron transfer energy (ETE) parameters were computed in gas phase and water. The results show that HAT would be the most favorable mechanism for explaining antioxidant activity of xanthones in gas phase, whereas the SPLET mechanism is thermodynamically favored in water.

Correlation of the antimicrobial activity of salicylaldehydes with broadening of the NMR signal of the hydroxyl proton. Possible involvement of proton exchange processes in the antimicrobial activity

Certain substituted salicylaldehydes are potent antibacterial and antifungal agents and some of them merit consideration as potential chemotherapeutic agents against Candida infections, but their mechanism of action has remained obscure. We report here a distinct correlation between broadening of the NMR signal of the hydroxyl proton of salicylaldehydes and their activity against several types of bacteria and fungi. When proton NMR spectra of the compounds were determined using hexadeuterodimethylsulfoxide as solvent and the height of the OH proton signal was measured, using the signal of the aldehyde proton as an internal standard, it was discovered that a prerequisite of potent antimicrobial activity is that the proton signal is either unobservable or relatively very low, i.e. that it is extremely broadened. Thus, none of the congeners whose OH proton signal was high were potent antimicrobial agents. Some congeners that gave a very low OH signal were, however, essentially inactive against the microbes, indicating that although drastic broadening of the OH signal appears to be a prerequisite, also other (so far unknown) factors are needed for high antimicrobial activity. Because broadening of the hydroxyl proton signal is related to the speed of the proton exchange process(es) involving that proton, proton exchange may be involved in the mechanism of action of the compounds. Further studies are needed to analyze the relative importance of different factors (such as electronic effects, strength of the internal hydrogen bond, co-planarity of the ring and the formyl group) that determine the rates of those processes.

Unraveling the mechanism of a reversible photoactivated molecular proton crane.

We study the structural dynamics of the photoactivated molecular proton crane 7-hydroxy-8-(morpholinomethyl)quinoline using femtosecond UV-pump IR-probe spectroscopy. Upon electronic excitation, a proton is transferred from the hydroxy to the amine group located on the rotatable morpholino side group. This morpholino group subsequently delivers the proton to the aromatic quinoline nitrogen by rotation around the C-C bond. Time-resolved vibrational spectroscopy allows us to study this process in unprecedented detail. We find that the transport of the proton involves multiple time scales. Upon photoexcitation, the OH proton is transferred within <300 fs to the morpholino side group. After this, the intramolecular hydrogen bond that locks the crane arm breaks with a time constant of 36 ± 1 ps. Subsequently, the protonated crane arm rotates with a time constant of 334 ± 12 ps to deliver the proton at the quinoline moiety. After the proton crane has returned to its electronic ground state with a time constant 700 ± 22 ps, the proton is transferred back from the quinoline nitrogen to the negatively charged O atom. The time constant of the back rotation is 39.8 ± 0.2 ns, about 200 times slower than the forward proton transfer.

Unconventional redox properties of hydroquinones with intramolecular OH−N hydrogen bonds

The redox (chemical and electrochemical) properties of several hydroquinones are reported in which the OH protons are engaged in intramolecular hydrogen bonds to a nitrogen-based acceptor (pyridine or amine). The 1,4-hydroquinones generally undergo reversible oxidation to quinones in which both OH protons have transferred to the pendant bases; the oxidation processes are generally chemically and electrochemically reversible, in stark contrast with normal hydroquinones, which are oxidized irreversibly (via proton loss) to quinones. The oxidation processes, believed to occur in concerted proton/electron transfer steps, are at much lower potentials for the hydrogen-bonded derivatives relative to unsubstituted derivatives. In contrast, isomeric 1,3-hydroquinones (resorcinols) are oxidized irreversibly at relatively high potentials. The stability of some of the 1,4-hydroquinone oxidized species permits their isolation and characterization both spectroscopically and structurally. Somewhat surprisingly, in the oxidized species in which the proton is now located on the nitrogen base, the characterization data indicate that there is no NH−O hydrogen bond. Relationships between the particulars of the redox properties of the hydroquinones (potentials, reversibility/stability) and molecular structure are discussed.

Intra- and intermolecular hydrogen bonding in acetylacetone and benzoylacetone derived enaminone derivatives

The structure and hydrogen bonding in solution of acetylacetone and benzoylacetone derived enaminone derivatives, 1a–1d and 2a–2d, have been studied by a combination of experimental (NMR and UV spectroscopies) and theoretical (PM6 and DFT) methods. It has been shown that all studied compounds predominantly existed in the localised keto-amine tautomeric form in solution as found also in the solid state. Significant line-broadening and down-field chemical shifts of NH and OH protons have strongly indicated that both groups formed hydrogen bonds, which has further been supported by quantum chemical calculations. Temperature and concentration dependent NMR measurements have pointed towards the fact that NH protons are engaged in strong intramolecular hydrogen bonds of the NH⋯OC type in both solvents. On the other hand, OH protons are involved in weaker intermolecular hydrogen bonding with solvent molecules in DMSO, while in chloroform intermolecular interactions between two molecules dominate. The results presented in this paper can be used for better understanding and further exploiting properties these ligands possess, especially their bioactivity.

1H NMR spectra of alcohols and diols in chloroform: DFT/GIAO calculation of chemical shifts.

Proton nuclear magnetic resonance (NMR) shifts of aliphatic alcohols in chloroform have been computed on the basis of density functional theory, the solvent being included by the integral-equation-formalism polarisable continuum model of Gaussian 09. Relative energies of all conformers are calculated at the Perdew, Burke and Ernzerhof (PBE)0/6-311+G(d,p) level, and NMR shifts by the gauge-including atomic orbital method with the PBE0/6-311+G(d,p) geometry and the cc-pVTZ basis set. The 208 computed CH proton NMR shifts for 34 alcohols correlate very well with the experimental values, with a gradient of 1.00 ± 0.01 and intercept close to zero; the overall root mean square difference (RMSD) is 0.08 ppm. Shifts for CH protons of diols in chloroform are well correlated with the theoretical values for (isotropic) benzene, with similar gradient and intercept (1.02 ± 0.01, -0.13 ppm), but the overall RMSD is slightly higher, 0.12 ppm. This approach generally gives slightly better results than the CHARGE model of Abraham et al. The shifts of unsaturated alcohols in benzene have been re-examined with Gaussian 09, but the overall fit for CH protons is not improved, and OH proton shifts are worse. Shifts of vinyl protons in alkenols are systematically overestimated, and the correlation of computed shifts against the experimental data for unsaturated alcohols follows a quadratic equation. Splitting the 20 compounds studied into two sets, and applying empirical scaling based on the quadratic for the first set to the second set, gives an RMSD of 0.10 ppm. A multi-standard approach gives a similar result.

Theoretical investigation on the bond dissociation enthalpies of phenolic compounds extracted from Artocarpus altilis using ONIOM(ROB3LYP/6-311++G(2df,2p):PM6) method

Theoretical calculations have been performed to predict the antioxidant property of phenolic compounds extracted from Artocarpus altilis. The OH bond dissociation enthalpy (BDE), ionization energy (IE), and proton dissociation enthalpy (PDE) of the phenolic compounds have been computed. The ONIOM(ROB3LYP/6-311++G(2df,2p):PM6) method is able to provide reliable evaluation for the BDE(OH) in phenolic compounds. An important property of antioxidants is determined via the BDE(OH) of those compounds extracted from A. altilis. Based on the BDE(OH), compound 12 is considered as a potential antioxidant with the estimated BDE value of 77.3kcal/mol in the gas phase.

1H-NMR as a structural and analytical tool of intra- and intermolecular hydrogen bonds of phenol-containing natural products and model compounds.

Experimental parameters that influence the resolution of 1H-NMR phenol OH signals are critically evaluated with emphasis on the effects of pH, temperature and nature of the solvents. Extremely sharp peaks (Δν1/2 ≤ 2 Hz) can be obtained under optimized experimental conditions which allow the application of 1H-13C HMBC-NMR experiments to reveal long range coupling constants of hydroxyl protons and, thus, to provide unequivocal assignment of the OH signals even in cases of complex polyphenol natural products. Intramolecular and intermolecular hydrogen bonds have a very significant effect on 1H OH chemical shifts which cover a region from 4.5 up to 19 ppm. Solvent effects on –OH proton chemical shifts, temperature coefficients (Δδ/ΔT), OH diffusion coefficients, and nJ(13C, O1H) coupling constants are evaluated as indicators of hydrogen bonding and solvation state of phenol –OH groups. Accurate 1H chemical shifts of the OH groups can be calculated using a combination of DFT and discrete solute-solvent hydrogen bond interaction at relatively inexpensive levels of theory, namely, DFT/B3LYP/6-311++G (2d,p). Excellent correlations between experimental 1H chemical shifts and those calculated at the ab initio level can provide a method of primary interest in order to obtain structural and conformational description of solute-solvent interactions at a molecular level. The use of the high resolution phenol hydroxyl group 1H-NMR spectral region provides a general method for the analysis of complex plant extracts without the need for the isolation of the individual components.

Multinuclear solid-state NMR characterization of the Brønsted/Lewis acid properties in the BP HAMS-1B (H-[B]-ZSM-5) borosilicate molecular sieve using adsorbed TMPO and TBPO probe molecules

The acid properties of a dehydrated borosilicate, HAMS-1B (H-[B]-ZSM-5), including the acid types, strengths, location, and quantities are investigated by means of trialkylphosphine oxides through multinuclear 1D/2D MAS NMR experiments. 11B DQF-STMAS combined with 1H MAS NMR studies revealed B–OH and distinct Si–OH protons associated with trigonal boron. 31P NMR spectra of TMPO-treated HAMS-1B reveal three Brønsted and three Lewis acid sites. We have found a number of limitations applying the TMPO/TBPO method to identify internal/external acidity. Therefore, we propose a new approach to unambiguously discriminate external/internal acid sites by treating a pore-free and pore-blocked HAMS-1B zeolite. This method provided unique structural insight regarding the identification of boron species/coordinations associated with Brønsted/Lewis acid sites. Additionally, ICP analysis in tandem with solid-state NMR enabled full assignment of the detected internal/external acid species and the study of their acid strength. Moreover, we identify the nature of TMPO complexes arising from Brønsted/Lewis interactions.

Curcumin derivatives as metal-chelating agents with potential multifunctional activity for pharmaceutical applications

Curcuminoids represent new perspectives for the development of novel therapeutics for Alzheimer's disease (AD), one probable mechanism of action is related to their metal complexing ability. In this work we examined the metal complexing ability of substituted curcuminoids to propose new chelating molecules with biological properties comparable with curcumin but with improved stability as new potential AD therapeutic agents. The K2T derivatives originate from the insertion of a -CH2COOC(CH3)3 group on the central atom of the diketonic moiety of curcumin. They retain the diketo-ketoenol tautomerism which is solvent dependent. In aqueous solution the prevalent form is the diketo one but the addition of metal ion (Ga3+, Cu2+) causes the dissociation of the enolic proton creating chelate complexes and shifting the tautomeric equilibrium towards the keto–enol form. The formation of metal complexes is followed by both NMR and UV–vis spectroscopy. The density functional theory (DFT) calculations on K2T21 complexes with Ga3+ and Cu2+ are performed and compared with those on curcumin complexes. [Ga(K2T21)2(H2O)2]+ was found more stable than curcumin one. Good agreement is detected between calculated and experimental 1H and 13C NMR data. The calculated OH bond dissociation energy (BDE) and the OH proton dissociation enthalpy (PDE), allowed to predict the radical scavenging ability of the metal ion complexed with K2T21, while the calculated electronic affinity (EA) and ionization potential (IP) represent yardsticks of antioxidant properties. Eventually theoretical calculations suggest that the proton-transfer-associated superoxide-scavenging activity is enhanced after binding metal ions, and that Ga3+ complexes display possible superoxide dismutase (SOD)-like activity.

1H NMR spectra of butane‐1,4‐diol and other 1,4‐diols: DFT calculation of shifts and coupling constants

The proton nuclear magnetic resonance (NMR) spectra of butane-1,4-diol, pentane-1,4-diol, (S,S)-hexane-2,5-diol, 2,5-dimethylhexane-2,5-diol and cyclohexane-1,4-diols (cis and trans) in benzene and some other solvents have been analysed. The conformer distribution and the NMR shifts of these diols in benzene have been computed on the basis of the density functional theory, the solvent being included by means of the integral-equation-formalism polarizable continuum model implemented in Gaussian 09. Relative Gibbs energies of all conformers are calculated at the Perdew, Burke and Ernzerhof (PBE)0/6-311+G(d,p) level and NMR shifts by the gauge-including atomic orbital method with the PBE0/6-311+G(d,p) geometry and the cc-pVTZ basis set. Vicinal three-bond coupling constants for the acyclic diols are calculated from the relative conformer populations, the geometries and generalized Karplus equations developed by Altona's group; these correlate well with the experimental values. The solvent dependence of coupling constants for butane-1,4-diol is attributed to conformational change. Coupling constants for the rigid cyclohexane-1,4-diols do not change with solvent and are readily explained in terms of their geometries. The NMR shifts of hydrogen-bonded protons in individual conformers of alkane-1,n-diols show a very rough correlation with the OH · · · OH distances. The computed overall NMR shifts for CH protons in 1,2-diols, 1,3-diols and 1,4-diols are systematically high but correlate very well with the experimental values, with a gradient of 1.07 ± 0.01; those for OH protons correlate less well.

Liquid state dynamic nuclear polarization of ethanol at 3.4 T (95 GHz)

Dynamic Nuclear Polarization (DNP) in the liquid state has become the focus of attention to improve the NMR sensitivity of mass limited samples. The Overhauser model predicts a fast reduction in DNP enhancement at high magnetic fields where the electron Larmor frequency exceeds the typical inverse correlation time of the magnetic interaction between an unpaired electron spin of a radical and proton spins of the solvent molecules. The Overhauser hard sphere model is able to predict quantitatively the DNP enhancement for water TEMPOL solutions. The increase in temperature due to dielectric heating of the sample acts to reduce the correlation times and allows a substantial Overhauser DNP. In this paper we extend the work done on water towards other small molecules, such as ethanol. Experimentally we observe a similar enhancement for all three proton groups in the ethanol molecule. The classical interpretation of the low field Overhauser experiments on ethanol invokes a very fast anisotropic rotation of the hydrogen bonded TEMPOL-ethanol complex to explain the fast relaxation of the OH proton. Here we will discuss W-band relaxation and DNP enhancement within this classical model. Although the description can be made quantitative, the invoked parameters do not seem to be realistic. We will propose an alternative model based on the dynamic interaction both in free collision and due to modulation of the hydrogen bond length of the complex.

Behavior of Polyhydric Alcohols at Ice/Liquid Interface

Glycerol (Gly) and 1,3-butanediol (13BD) are widely used as a cryoprotective reagent. Although these compounds have similar chemical structures, their interfacial behavior is not necessarily the same. Frozen aqueous 13BD gives discrete liquid inclusions dispersed in an ice matrix, whereas frozen aqueous Gly forms the film-like liquid phase spread over the entire ice crystal surface. Nuclear magnetic resonance (NMR) measurements of Gly/water at 253 K indicate that the chemical shifts of both water and alcohol OH protons move upfield with increasing concentration of Gly regardless of whether the solution is frozen (<48 wt % Gly) or not (≥48 wt % Gly). In contrast, the chemical shifts remain constant over the entire concentration range for 13BD. Gly forms a molecularly homogeneous mixture with water in any ratio and also interacts with ice similar to liquid water; this leads to the continuous changes in the chemical shifts. However, 13BD and water mainly form individual clusters, which weakly interact with i...

Association of 2-acylaminopyridines and benzoic acids. Steric and electronic substituent effect studied by XRD, solution and solid-state NMR and calculations

Eight single crystal X-ray structures, solid-state NMR spectroscopic, and theoretical studies utilizing QTAIM methodology were used to characterize the 2-acyl (alkyl in acyl=methyl, ethyl, t-butyl, and 1-adamantyl) amino-6-R-pyridine/4-R′-benzoic acid (R,R′=H or Me) cocrystals. As expected among alkyl groups 1-adamantyl due to its bulkiness has the most significant effect on the relative positions of molecules in cocrystals. In addition, the subtle electronic and steric effects by the methyl substituents were observed. The theoretical calculations with full geometry optimizations are in agreement with the experimental findings (geometry, energy of hydrogen bonds). Based on the crystal structures and calculations it is concluded that p-methyl substituent in benzoic acid increase the hydrogen bond accepting ability of the CO oxygen and decreases the hydrogen bond donating ability of OH proton. The 15N solid-state (CP MAS) NMR chemical shifts prove that molecules in cocrystal are held together by hydrogen bonding. The biggest variation in the 15N chemical shift of acylamino nitrogen can be related with the size of the alkyl group in acyl moiety.