Measurement of longitudinal relaxation times using surface coils

The poor B₁ field homogeneity associated with surface coils reduces the effectiveness of inversion-recovery techniques for determination of T₁ relaxation times. This paper presents a variation of the saturation-recovery T₁ experiment which introduces periodic phase shifts in the saturating irradiation to achieve rapid and effective saturation of the sample magnetization, thereby avoiding the complications of B₁ field inhomogeneity. Comparison of the presented technique with the inversion-recovery technique utilizing a composite inverting pulse and alternating phase acquisition is provided. A discussion of the relative merits of each technique is presented.     

Pulsed field gradients in simulations of one- and two-dimensional NMR spectra

A method for the inclusion of the effects of z-axis pulsed field gradients in computer simulations of an arbitrary pulsed NMR experiment with spin (1/2) nuclei is described. Recognizing that the phase acquired by a coherence following the application of a z-axis pulsed field gradient bears a fixed relation to its order and the spatial position of the spins in the sample tube, the sample is regarded as a collection of volume elements, each phase-encoded by a characteristic, spatially dependent precession frequency. The evolution of the sample's density matrix is thus obtained by computing the evolution of the density matrix for each volume element. Following the last gradient pulse, these density matrices are combined to form a composite density matrix which evolves through the rest of the experiment to yield the observable signal. This approach is implemented in a program which includes capabilities for rigorous inclusion of spin relaxation by dipole-dipole, chemical shift anisotropy, and random field mechanisms, plus the effects of arbitrary RF fields. Mathematical procedures for accelerating these calculations are described. The approach is illustrated by simulations of representative one- and two-dimensional NMR experiments.     

Measurement of longitudinal and rotating frame relaxation times through fully J-decoupled homonuclear spectra.

It is shown that fully J-decoupled homonuclear spectra involving Lorentzian lines can be readily obtained by straightforward processing of the 2D data arising from a conventional spin echo sequence (pi/2-t(1)/2-pi-t(1)/2-Acq(t(2))) used in the so-called J-resolved experiment. The method simply rests on power spectra with the drawback of lines having meaningless relative intensities. In principle, the experiment should also yield transverse relaxation times. Several tests demonstrate that this is not so, due to pulse imperfections and nonresolved long-range J couplings. Conversely, longitudinal and rotating frame relaxation times can be easily determined by means of an appropriate preparation period (for instance, a saturation-recovery period in the case of longitudinal relaxation) inserted before the 2D spin echo sequence. Since one is dealing with a single line per nucleus, relaxation measurements become reliable and accurate.     

Picoliter (1)H NMR spectroscopy

In this study, a 267-microm-diameter solenoid transceiver is used to acquire localized (1)H NMR spectra and the measured signal-to-noise ratio (SNR) at 500 MHz is shown to be within 20--30% of theoretical limits formulated by considering only its resistive losses. This is illustrated using a 100-microm-diameter globule of triacylglycerols (approximately 900mM) that may be an oocyte precursor in young Xenopus laevis frogs and a water sample containing choline at a concentration often found in live mammalian cells (approximately 33 mM). In chemical shift imaging (CSI) experiments performed using a few thousand total scans, the choline methyl line is shown to have an acceptable SNR in resolved volume elements containing only 50 pL of sample, and localized spectra are resolved from just 5 pL in the Xenopus globule. These findings demonstrate the feasibility of performing (1)H NMR on picoliter-scale sample volumes in biological cells and tissues and illustrate how the achieved SNR in spectroscopic images can be predicted with reasonable accuracy at microscopic spatial resolutions.     

Double quantum filtered (1)H NMR spectroscopy enables quantification of lactate in muscle

In this study we address the question of quantification of muscle lactate using double quantum filtered (DQF) (1)H NMR spectroscopy where dipolar and scalar coupled spectra are acquired. For this, lactate content in muscle samples was independently determined using a conventional enzymatic assay and DQF, (1)H NMR spectroscopy. NMR quantification of lactate relied on comparison of muscle spectra with similarly acquired spectra of standard lactate solutions. Transverse relaxation, T(2), and dipolar coupling effects were investigated at two different orientations of muscle fibers relative to B(o) and at various lactate concentrations. In all cases, we found a biexponential T(2) decay of the lactate methyl signal with a long T(2) of 142 ms (+/-8 ms, n=24) and a short T(2) of 37 ms (+/-6 ms, n=24). Lactate content of muscle determined by NMR spectroscopy agreed with the results obtained from enzymatic assays of the same samples provided that T(2) effects as well as the presence of both scalar and dipolar coupling interactions of lactate in muscle were taken into account.     

Internal dynamics and hydration of solid proteins by1H NMR relaxation and FTIR spectroscopy

Temperature dependences of¹H nonselective nuclear magnetic resonanceT ₁ andT ₂ relaxation times measured at 27 MHz have been studied on solid human serum albumin (HSA) samples at various hydrations. The data were interpreted in terms of three kinds of internal motions in a protein and microdynamic parameters of the motions were obtained by a “model-free” approach. Two fast motions with correlation times lying in the range of tens to hundreds picoseconds were shown to be essentially insensitive to hydration. Unlike lysozyme and bovine albumin, HSA reveals relaxation transition due to slow motion in the room temperature range thus allowing one to obtain microdynamic parameters more precisely. Hydration leads to a shortening of the correlation time from hundreds to tens nanoseconds and to a less restricted movement. The comparison of the hydration dependence of relaxation parameters with infrared spectra of HSA side chain groups clearly shows that methyl protons are evidently involved in a slow motion, following the saturation of the protein globule surface by water. The same dependence correlating with solvent accessible surface areas was shown to exist for some other proteins. In addition to the main set of protons performing a solidlike movement, a small amount of much more mobile protons is also present with its proportion rising steeply with hydration and temperature. The origin of these protons is discussed.     

Measurement of homonuclear (2)J-couplings from spin-state selective double-/zero-quantum two-dimensional NMR spectra.

(1)H-detected two-dimensional double-/zero-quantum experiments are described for measurement of homonuclear (2)J(HH)-couplings of NH(2) or CH(2) groups in proteins. These experiments utilize multiple-quantum coherence for determination of the size and the absolute sign of the geminal scalar and dipolar couplings in the presence of broad lines. Spectra are simplified by gradient selection and spin-state selective filters.     

Dynamic disorder in solid tetrakis(trimethylstannyl)methane, C(SnMe3)4, investigated by one- and two-dimensional variable-temperature119Sn and13C NMR spectroscopy

Modes of molecular reorientation in solid tetrakis(trimethylstannyl)methane, C(SnMe₃)₄, have been investigated by one- and two-dimensional¹³C and¹¹⁹Sn static and magic-angle spinning nuclear magnetic resonance spectroscopy (NMR) in the temperature range from 150 to 290 K. Spectral lineshape fitting of one- and two-dimensional¹¹⁹Sn NMR experiments shows the pseudo-fivefold disorder previously observed by single-crystal X-ray diffraction on C(SnMe₃)₄ to be dynamic disorder (activation energy E_a ? 32 kJmol^?1). A dynamic-disorder model where each tin atom in a C(SnMe₃)₄ molecule occupies the twenty sites of a nearly perfect pentagonal dodecahedron with equal probability agrees best with the experimental solid-state NMR results.     

Complete NMR fingerprints of proteins in H2O solution without solvent presaturation. An application of two-dimensional longitudinal two-spin-order spectroscopy

A method for obtaining complete two-dimensional ¹H NMR amide proton-C_α proton J-connectivity maps for proteins (the “fingerprint”) is describe. The method relies on the conversion of antiphase single-quantum coherence into longitudinal two-spin order which is made observable by utilizing a selective solvent-suppression readout pulse. Since irradiation of the solvent resonance is unnecessary in this experiment, J connectivities are observable even for protons resonating exactly at the solvent frequency. Thus, a complete protein fingerprint is obtained at a single set of experimental conditions. The method is demonstrated for a 75-amino-acid protein.     

Mobility and structure of human serum albumin powders in water-acetonitrile mixtures by1H NMR relaxation and FTIR spectroscopy

The effect of acetonitrile on protein dynamics was investigated for solid human serum albumin samples at various hydration levels. Temperature dependences of¹H nonselective nuclear magnetic resonanceT ₁ andT ₂ relaxation times at 27 MHz have been measured and data were interpreted in terms of three kinds of internal motions in the protein. Microdynamic parameters of the motions were obtained within a “model-free” approach. It was found that acetonitrile hardly affects the fast motions but noticeably influences the slow motion of side chain groups, shortening the correlation time and increasing the amplitude of the motion. The acetonitrile effect on dynamics is likely based on the appearance of additional free volume as a result of the formation of rigid helical parts in the protein structure. Water, plasticizing the protein structure, promotes the action of organic solvent. A definite part of side chain groups, slowly moving in the same frequency window as the rest of protein side chain groups, performs less constrained “liquidlike” motion. The relative population of these highly movable protons is closely correlated with the increment of the helical structure induced by water and acetonitrile.     

Three-dimensional protein NMR TROSY-type (15)N-resolved (1)H(N)-(1)H(N) NOESY spectra with diagonal peak suppression

An earlier two-dimensional NOESY experiment with diagonal peak suppression in the (1)H(N)-(1)H(N) region is extended to three dimensions by including (15)N evolution while maintaining the TROSY approach throughout. The technique suppresses all anti-TROSY resonances by appropriate pulse sequence elements and for large molecules at high fields possible semi- and anti-TROSY artifacts are further suppressed by virtue of much shorter transverse relaxation times for these components. The new technique is demonstrated using an (15)N-labeled protein sample, RAP 17-97 (N-terminal domain of alpha2-macroglobulin Receptor Associated Protein), in H(2)O at 500 MHz.     

Strong spin-spin coupling in the two-dimensional J-resolved 360-MHz 1H NMR spectra of the common amino acids

The two-dimensional J-resolved ^H NMR spectra at 360 MHz of the 20 common amino acids have been investigated. The characteristic features of strong coupling are described. Advantages and drawbacks of projections, cross sections, and contour plots for the presentation of spectra including strong coupling are illustrated with selected examples. On the basis of the amino acid data practical aspects of the use of two-dimensional J-resolved spectroscopy to improve the resolution of high-field ¹H NMR spectra of proteins are discussed.     

Gated spin relaxation in (1 1 0)-oriented quantum wells

Growth, photoluminescence characterisation and time-resolved optical measurements of electron spin dynamics in (1 1 0)-oriented GaAs/AlGaAs quantum wells are described. Conditions are given for MBE growth of good-quality quantum wells, judged by the width of low-temperature excitonic photoluminescence. At 170 K the electron spin relaxation rate in (1 1 0)-oriented wells shows a 100-fold reduction compared to equivalent (1 0 0)-oriented wells and also a 10-fold increase with applied electric field from 20 to 80 kV cm⁻¹. There is evidence for similar dramatic effects at 300 K. Spin relaxation is field independent below 20 kV cm⁻¹ reflecting quantum well asymmetry. The results indicate the achievability of voltage-gateable quantum well spin memory time longer than 10 ns at room temperature simultaneously with high electron mobility.     

T(1)--T(2) correlation spectra obtained using a fast two-dimensional Laplace inversion

Spin relaxation is a sensitive probe of molecular structure and dynamics. Correlation of relaxation time constants, such as T(1) and T(2), conceptually similar to the conventional multidimensional spectroscopy, have been difficult to determine primarily due to the absense of an efficient multidimensional Laplace inversion program. We demonstrate the use of a novel computer algorithm for fast two-dimensional inverse Laplace transformation to obtain T(1)--T(2) correlation functions. The algorithm efficiently performs a least-squares fit on two-dimensional data with a nonnegativity constraint. We use a regularization method to find a balance between the residual fitting errors and the known noise amplitude, thus producing a result that is found to be stable in the presence of noise. This algorithm can be extended to include functional forms other than exponential kernels. We demonstrate the performance of the algorithm at different signal-to-noise ratios and with different T(1)--T(2) spectral characteristics using several brine-saturated rock samples.     

Fast and simultaneous measurement of longitudinal and transverse NMR relaxation times in a single continuous wave free precession experiment

The purpose of this communication is to describe a method for rapid and simultaneous determination of longitudinal (T1) and transversel (T2) relaxation times, based on a single continuous wave free precession (CWFP) experiment which employs RF pulses with a pi/2 flip angle. We analyze several examples, involving nuclei such as 1H, 31P, and 19F, where good agreement with T1 and T2 measurements obtained by traditional methods is apparent. We also compare with the more time-consuming steady-state free precession (SSFP) method of Kronenbitter and Schwenk where several experiments are needed to determine the optimum flip angle. The role of an inhomogeneous magnetic field on the observed decays and its effect upon the accuracy of relaxation times obtained by these methods is examined by comparing numerical simulations with experimental data. Possible sources of error and conditions to minimize its effects are described.