Solid-state F NMR investigation of poly(vinylidene fluoride) with high-power proton decoupling

The use of high-power proton decoupling has enabled highly-resolved spectra of fluorine polymers to be recorded, as is exemplified herein for semicrystalline poly(vinylidene fluoride) (PVDF). By means of high MAS speeds (up to 17 kHz), the spinning sidebands are removed from the whole of the relevant chemical shift range. For spectra of the crystalline regions of the polymer, the high-power decoupling is necessary, though its effect is not large. Various relaxation techniques have been used to examine the semicrystallinity and the polymorphism of PVDF, with special pulse sequences used to discriminate between the various domains. Different chemical shifts have been observed for the signals of the amorphous and crystalline phases. Those of the more immobile parts cover a substantial range.     

Homonuclear zero-quantum recoupling in fast magic-angle spinning nuclear magnetic resonance

Solid-state magic-angle-spinning NMR pulse sequences which implement zero-quantum homonuclear dipolar recoupling are designed with the assistance of symmetry theory. The pulse sequences are compensated on a short time scale by the use of composite pulses and on a longer time scale by the use of supercycles. (13)C dipolar recoupling is demonstrated in powdered organic solids at high spinning frequencies. The new sequences are compared to existing pulse sequences by means of numerical simulations. Experimental two-dimensional magnetization exchange spectra are shown for [U-(13)C]-L-tyrosine.     

Fluorine-19 solid state NMR study of vinylidenefluoride polymers using selective relaxation filters

Two fluoropolymers, poly(vinylidenefluoride) (PVDF) and a vinylidenefluoride telomer (VDFT), with molecular weights of 1 x 10(6) and 2 x 10(3) Da by GPC, respectively, have been analysed by 19F solid-state nuclear magnetic resonance (NMR) spectroscopy. Relaxation-filtered proton-decoupled magic-angle spinning (MAS) experiments, namely T1rho filter, dipolar filter (DF), direct-polarisation delayed acquisition (DPDA) and discrimination induced by variable-amplitude minipulses (DIVAM), allowed signals in the direct polarisation (DP) spectra of PVDF and the VDFT to be discussed in terms of rigid and mobile domains. Both samples showed signals, which were multi-componential, but they differ in the nature of the crystalline form present. Thus, the Vinylidenefluoride (VDF) telomer exhibited a crystalline component corresponding to beta PVDF, whereas the PVDF contained crystallites of the alpha form. Signals relating to end groups and reverse units, plus an anomalous signal displaying long-time transverse relaxation in the DPDA spectrum, were found for both polymers, though they showed diversity in chemical shift and content. Signals related to reverse units and/or end groups were seen between approximately -115 and approximately -117 ppm for both samples. High-speed MAS at higher magnetic field resulted in an increase in resolution so that signals previously attributed to single-phase characteristics are shown to indicate the possibility of several different mobilities. The results are debated with respect to molecular weight and relaxation parameters.     

Cross- and axial-peak intensities in 2D-SLF experiments based on cross-polarization--the role of the initial density matrix

Simulations and experiments on simple oriented systems have been used to estimate the relative ratio of cross-peak to axial-peak intensities in 2D-SLF experiments based on dipolar oscillations during cross-polarization (CP). The density matrix prior to dipolar evolution is considered and for an isolated spin pair, it is shown that direct calculations of the ratios match well with simulations and experimental results. Along with the standard CP pulse sequence, two other pulse sequences namely CP with polarization inversion (PI-CP) and another novel variation of the standard CP experiment (EXE-CP) reported recently have been considered. Inclusion of homonuclear dipolar coupling has been observed to increase the axial-peak intensities. In combination with Lee-Goldburg (LG) decoupling, experiments on an oriented liquid crystalline sample have been carried out and the performance of the pulse schemes have been compared. The applicability of the new pulse sequence for different samples and different nuclei is discussed. Such studies are expected to lead to a better understanding of the experiments and to the design of useful pulse sequences.     

Sensitivity-enhanced IPAP experiments for measuring one-bond C–C and C–H residual dipolar couplings in proteins

Sensitivity-enhanced 2D IPAP experiments using the accordion principle for measuring one-bond 13C'-13Calpha and 1Halpha-13Calpha dipolar couplings in proteins are presented. The resolution of the resulting spectra is identical to that of the decoupled HSQC spectra and the sensitivity of the corresponding 1D acquisitions are only slightly lower than those obtained with 3D HNCO and 3D HN(COCA)HA pulse sequences due to an additional delay 2Delta. For cases of limited resolution in the 2D 15N-1HN HSQC spectrum the current pulse sequences can easily be modified into 3D versions by introducing a poorly digitized third dimension, if so desired. The experiments described here are a valuable addition to the suites available for determination of residual dipolar couplings in biological systems.     

Sensitivity-enhanced IPAP experiments for measuring one-bond 13C'-13Calpha and 13Calpha-1Halpha residual dipolar couplings in proteins

Sensitivity-enhanced 2D IPAP experiments using the accordion principle for measuring one-bond 13C'-13Calpha and 1Halpha-13Calpha dipolar couplings in proteins are presented. The resolution of the resulting spectra is identical to that of the decoupled HSQC spectra and the sensitivity of the corresponding 1D acquisitions are only slightly lower than those obtained with 3D HNCO and 3D HN(COCA)HA pulse sequences due to an additional delay 2Delta. For cases of limited resolution in the 2D 15N-1HN HSQC spectrum the current pulse sequences can easily be modified into 3D versions by introducing a poorly digitized third dimension, if so desired. The experiments described here are a valuable addition to the suites available for determination of residual dipolar couplings in biological systems.     

Adjusting Conditions for High-resolution Pulsed NMR in Solids

Adjusting conditions of pulse sequences for high-resolution nmr in solids are demonstrated by the example of the WHH4 cycle. For this pulse sequence the well-known adjusting condition [1] occurs as limiting case at sufficiently weak dipolar interaction. The relations between the pulse rotation angle and interval time proved to be dependent on the dipolar interaction. Therefore it is impossible to suppress the dipolar interaction in ordinary solids completely. If the strength of the dipolar interaction varies in a range depending on the allowable inhomogeneity of the rf field a spectrometer for high-resolution nmr in solids can be adjusted in an almost optimal way.     

MRI with the dipolar interaction refocusing techniques: analysis of the effectiveness for the solid-state polymers

The effectiveness of solid-echo and magic-echo phase-encoding solid-state magnetic resonance imaging methods was tested to determine possible improvement of sensitivity and spatial resolution for investigation of various types of solid polymers. The dipolar interaction refocusing pulse sequences have been used to elongate the possible phase-encoding period and to improve the signal sensitivity. The comparison of both dipolar refocusing techniques with conventional single point imaging method was made. The optimization of the phase-encoding time and magnetization recovery periods were performed basing on (1)H spectra and longitudinal relaxation measurements, respectively. The influence of imaging artifacts (intrinsic for each technique) on image quality was investigated. The effectiveness of the artifacts suppression methods was tested.     

Structure determination in "shiftless" solid state NMR of oriented protein samples

An efficient formalism for calculating protein structures from oriented-sample NMR data in the torsion-angle space is presented. Angular anisotropies of the NMR observables are treated by utilizing an irreducible spherical basis of rotations. An intermediate rotational transformation is introduced that greatly speeds up structural fitting by rendering the dependence on the torsion angles Φ and Ψ in a purely diagonal form. Back-calculation of the simulated solid-state NMR spectra of protein G involving 15N chemical shift anisotropy (CSA), and 1H-15N and 1Hα-13Cα dipolar couplings was performed by taking into account non-planarity of the peptide linkages and experimental uncertainty. Even a relatively small (to within 1 ppm) random variation in the CSA values arising from uncertainties in the tensor parameters yields the RMSD's of the back-calculated structures of more than 10 ?. Therefore, the 15N CSA has been substituted with heteronuclear dipolar couplings which are derived from the highly conserved bond lengths and bond angles associated with the amino-acid covalent geometry. Using the additional 13Cα-15N and 13C'-15N dipolar couplings makes it possible to calculate protein structures entirely from "shiftless" solid-state NMR data. With the simulated "experimental" uncertainty of 15 Hz for protein G and 120 Hz for a helical hairpin derived from bacteriorhodopsin, back-calculation of the synthetic dipolar NMR spectra yielded a converged set of solutions. The use of distance restraints dramatically improves structural convergence even if larger experimental uncertainties are assumed.     

Pulse error compensating symmetric magic-echo trains

We present improved line-narrowing sequences for dipolar coupled spin systems, based on a train of magic-echoes which are compensated for the effects of finite pulse widths and utilize symmetry properties of supercycles. Sequences are introduced for spectroscopy and imaging by proper choice of a phase alternating scheme. Using a 16 pulse time-suspension magic-echo cycle, the highest level of line-narrowing achieved was 2.7 Hz for the [100] direction of a single crystal of calcium fluoride, a reduction in linewidth by 4 orders of magnitude.     

NMR imaging of solids by Jeener-Broekaert phase encoding

A method for imaging of spectral parameters of solid samples is presented. The detected NMR signals are Jeener-Broekaert echoes. No read field gradient is applied during the acquisition, so that wide-line spectral parameters can be evaluated and be transferred to image contrasts. On the other hand, multipulse line-narrowing sequences can be applied during the echoes in order to obtain high-resolution spectra. The imaging principle is a pure phase-encoding Fourier technique in two or three dimensions. The phase-encoding gradients are active in the interval between the first two pulses of the Jeener-Broekaert three-pulse sequence. Between the second and the third pulse, the information is conserved in the dipolar (or quadrupolar) order state which is insensitive to field gradients and governed by the relatively slow dipolar (or quadrupolar) relaxation. This interval therefore can be chosen to be long enough to switch the gradients off. The third pulse “reads” the information of the spin-state order and produces an echo under homogeneous field conditions. In the case of two-dimensional imaging, a slice is preselected prior to the whole Jeener-Broekaert sequence by the aid of a LOSY slice-selection pulse. Test experiments are reported, and applications to polymer and biological materials are discussed.     

Pulse train measurement of ferroelectric switching in thin films of vinylidene fluoride/trifluoroethylene copolymer

Ferroelectric switching processes in thin films of a vinylidene fluoride/trifluoroethylene copolymer are investigated by pulse train measurement that involves the application of a train of unipolar on–off electric field pulses and the analysis of charge response. It is found that polarization reversal is considerably delayed as pulse width becomes shorter than the conventional switching time, but is eventually completed even if pulse width is reduced to 0.2 μs. The charge response in each on–off cycle was resolved into dielectric and ferroelectric components. The former is nonlinear as well as relaxational and reflects changes in dipolar fluctuation and orientation during polarization reversal. The latter is responsible for polarization reversal that progresses forward and backward depending on the on–off period. Net polarization reversal occurs first gradually and then rapidly, corresponding to nucleation and growth, respectively. The switching current responsible for these processes can be reproduced by the sum of power and lognormal functions. The implications of these results are discussed in relation to the switching process that occurs as a result of the rotation of chain molecules. PACS 42.70.Jk; 77.80.Fm; 77.22.Ej     

19F/29Si rotational-echo double-resonance and heteronuclear spin counting under fast magic-angle spinning in fluoride-containing octadecasil

19F/29Si rotational-echo double-resonance (REDOR) and theta-REDOR NMR techniques have been applied under fast magic-angle spinning to a powder sample of fluoride-containing octadecasil. Efficient dipolar recoupling was observed and the effect of finite pulse lengths was found to be negligible using standard radiofrequency field strengths. Moreover, the determined internuclear distance of the 19F-29Si spin pairs formed by the silicons in the D4R units (T-1 site) and the fluoride anions is in very good agreement with previous REDOR and Hartmann-Hahn cross-polarization measurements. Numerical simulation of the REDOR dephasing curves at both the T-1 and T-2 sites considering all fluoride anions in the infinite solid lattice clearly confirm the X-ray crystal structure of octadecasil. Heteronuclear spin-counting theta-REDOR experiments are found to be very useful to obtain direct insight into the local network of dipolar interactions. Indeed, while 19F-29Si pair-like behavior is confirmed at the T-1 site, multiple dipolar interactions are clearly evidenced at the T-2 site.     

Multiple pulse NMR imaging of polymers and chemistry.

Multiple pulse line narrowing techniques can be used to improve resolution and sensitivity in solid state NMR imaging. For example, pulse sequences which remove homonuclear dipolar broadening have been used to image proton-containing materials. Further enhancements in resolution and sensitivity are obtained by removing inhomogeneous interactions such as chemical shift, susceptibility, and heteronuclear dipolar broadening. Pulse sequences have been designed which provide efficient line narrowing over large spectral widths by taking into account the experimenter's control over the amplitude and time dependence of the gradient-induced resonance offset. These methods have been applied to centimeter sized samples to obtain images of polymers, composite materials, and gas-solid chemical reactions. T1 and T2 contrast allows differentiation between materials.     

Heteronuclear intermolecular single-quantum coherences in liquid nuclear magnetic resonance

This paper analyses the heteronuclear Cosy Revamped by Asymmetric Z-gradient Echo Detection pulse sequence. General theoretical expressions of the pulse sequence with arbitrary flip angles were derived by using dipolar field treatment and signals originating from heteronuclear intermolecular single-quantum coherences （iSQCs） in highly-polarized two spin-1/2 systems were mainly discussed in order to find the optimal flip angles. The results show that signals from heteronuclear iSQCs decay slower than those from intermolecular double-quantum coherences or intermolecular zero-quantum coherences. Magical angle experiments validate that the signals are from heteronuclear iSQCs and insensitive to the imperfection of radio-frequency flip angles. All experimental observations are in excellent agreement with theoretical predictions. The quantum-mechanical treatment leads to similar predictions to the dipolar field treatment.     

High resolution 2D 1H-13C correlation of cholesterol in model membrane

High resolution 2D NMR MAS spectra of liposomes, in particular 1H-13C chemical shifts correlations have been obtained on fluid lipid bilayers made of pure phospholipids for several years. We have investigated herein the possibility to obtain high resolution 2D MAS spectra of cholesterol embedded in membranes, i.e. on a rigid molecule whose dynamics is characterized mainly by axial diffusion without internal segmental mobility. The efficiency of various pulse sequences for heteronuclear HETCOR has been compared in terms of resolution, sensitivity and selectivity, using either cross polarization or INEPT for coherence transfer, and with or without MREV-8 homonuclear decoupling during t1. At moderately high spinning speed (9 kHz), a similar resolution is obtained in all cases (0.2 ppm for 1H(3,4), 0.15 ppm for 13C(3,4) cholesterol resonances), while sensitivity increases in the order: INEPT < CP(x4) < CP + MREV. At reduced spinning speed (5 kHz), the homonuclear dipolar coupling between the two geminal protons attached to C(4) gives rise to spinning sidebands from which one can estimate a H-H dipolar coupling of 10 kHz which is in good agreement with the known dynamics of cholesterol in membranes.     

The polymorphism of poly(vinylidene fluoride) V. The effect of hydrostatic pressure on the melting behavior of copolymers of vinylidene fluoride

The high-pressure melting behavior of samples of vinylidene fluoride copolymerized with vinyl fluoride, trifluoroethylene, or tetrafluoroethylene indicate that the copolymers have a lower entropy of melting than the poly(vinylidene fluoride) homopolymers in the same phase. As the comonomer size increases, the entropy of melting decreases. High-pressure crystallization of copolymers of 91-9 mole% vinylidene fluoride-trifluoroethylene and 93-7 mole% vinylidene fluoride-tetrafluoroethylene results in a high-melting form of phase I (planar zig-zag).     

Two Simple NMR Experiments for Measuring Dipolar Couplings in Asparagine and Glutamine Side Chains

Residual dipolar couplings are now widely used for structure determination of biological macromolecules. Until recently, the main focus has been on measurement of dipolar couplings in the protein main chain. However, with the aim of more complete protein structure, it is also essential to have information on the orientation of protein side chains. In addition, residual dipolar couplings can potentially be employed to study molecular dynamics. In this Communication, two simple NH(2) and spin-state edited experiments are presented for rapid and convenient determination of five residual dipolar couplings from (15)N, (1)H correlation spectrum in asparagine and glutamine side chains. The pulse sequences are demonstrated on two proteins, 30.4-kDa Cel6A in diluted liquid crystal phase and 18-kDa human cardiac troponin C in water.     

Spin diffusion of correlated two-spin states in a dielectric crystal

Reciprocal space measurements of spin diffusion in a single crystal of calcium fluoride (CaF2) have been extended to dipolar ordered states. The experimental results for the component of the spin diffusion rate parallel to the external field are D(parallel)(D)=29+/-3x10(-12) cm(2)/s for the [001] direction and D(parallel)(D)=33+/-4x10(-12) cm(2)/s for the [111] direction. The measured diffusion rates for dipolar order are faster than those for Zeeman order and are considerably faster than predicted by simple theoretical models. It is suggested that constructive interference in the transport of the two-spin states is responsible for this enhancement. As expected, the anisotropy in the diffusion rates is observed to be significantly less for dipolar order compared to the Zeeman case.     

Measurement of small one-bond proton-carbon residual dipolar coupling constants in partially oriented (13)C natural abundance oligosaccharide samples: analysis of heteronuclear (1)J(CH)-modulated spectra with the BIRD inversion pulse

Two 2D J-modulated HSQC-based experiments were designed for precise determination of small residual dipolar one-bond carbon-proton coupling constants in (13)C natural abundance carbohydrates. Crucial to the precision of a few hundredths of Hz achieved by these methods was the use of long modulation intervals and BIRD pulses, which acted as semiselective inversion pulses. The BIRD pulses eliminated effective evolution of all but (1)J(CH) couplings, resulting in signal modulation that can be described by simple modulation functions. A thorough analysis of such modulation functions for a typical four-spin carbohydrate spin system was performed for both experiments. The results showed that the evolution of the (1)H-(1)H and long-range (1)H-(13)C couplings during the BIRD pulses did not necessitate the introduction of more complicated modulation functions. The effects of pulse imperfections were also inspected. While weakly coupled spin systems can be analyzed by simple fitting of cross peak intensities, in strongly coupled spin systems the evolution of the density matrix needs to be considered in order to analyse data accurately. However, if strong coupling effects are modest the errors in coupling constants determined by the "weak coupling" analysis are of similar magnitudes in oriented and isotropic samples and are partially cancelled during dipolar coupling calculation. Simple criteria have been established as to when the strong coupling treatment needs to be invoked.