Assessment of chemical and biochemical stabilization of organic C in soils from the long-term experiments at Rothamsted (UK).

Biological and chemical stabilization of organic C was assessed in soils sampled from the long-term experiments at Rothamsted (UK), representing a wide range of carbon inputs and managements by extracting labile, non-humified organic matter (NH) and humic substances (HS). Four sequentially extracted humic substances fractions of soil organic matter (SOM) were extracted and characterized before and after a 215-day laboratory incubation at 25 degrees C from two arable soils, a woodland soil and an occasionally stubbed soil. The fractions corresponded to biochemically stabilised SOM extracted in 0.5M NaOH (free fulvic acids (FA) and humic acids (HA)) and chemically plus biochemically stabilised SOM extracted from the residue with 0.1M Na4P2O7 plus 0.1M NaOH (bound FA and HA). Our aim was to investigate the effects of chemical and biochemical stabilization on carbon sequestration. The non-humic to humic (NH/H) C ratio separated the soils into two distinct groups: arable soils (unless fertilised with farmyard manure) had an NH/H C ratio between 1.05 and 0.71, about twice that of the other soils (0.51-0.26). During incubation a slow, but detectable, decrease in the NH/H C ratio occurred in soils of C input equivalent or lower to 4Mgha(-1)y(-1), whereas the ratio remained practically constant in the other soils. Before incubation the free to bound humic C ratio increased linearly (R2=0.91) with C inputs in the soils from the Broadbalk experiment and decreased during incubation, showing that biochemical stabilization is less effective than chemical stabilization in preserving humic C. Changes in delta13C and delta15N after incubation were confined to the free FA fractions. The delta13C of free FA increased by 1.48 and 0.80 per thousand, respectively, in the stubbed and woodland soils, indicating a progressive biological transformation. On the contrary, a decrease was observed for the bound FA of both soils. Concomitantly, a Deltadelta15N of up to +3.52 per thousand was measured after incubation in the free FA fraction and a -2.58 Deltadelta15N in the bound FA. These changes, which occurred during soil incubation in the absence of C inputs, indicate that free FA fractions were utilised by soil microorganisms, and bound FA were decomposed and replaced, in part, by newly synthesized FA. The 13CPMAS-TOSS NMR spectra of free HA extracted before and after 215 days of incubation were mostly unchanged. In contrast, changes were evident in bound HA and showed an increase in aromatic C after incubation.

The first proton NMR imaging of ice: stray-field imaging and relaxation studies.

A proton magnetic resonance image of ice was observed with the stray-field (STRAFI) technique. A preliminary study of proton relaxation times was performed in water and ice, at different temperatures. For example, a value of 3.5 micros for the spin-spin relaxation time, T(2), was found in ice at 258 K. Such a short T(2) value leads to significant signal loss, as compared to liquid water, and to a shortening of the STRAFI echo-trains. In particular, a STRAFI signal for protons in ice could be observed only at echo times as short as 15 and 25 micros, for RF pulse durations corresponding to 90 degrees and 50 degrees magnetisation tip angles, respectively. This behaviour is in contrast with that of deuteriated water. Imaging ice, as shown here, opens new prospects in studies involving environmental and materials science, for example.

Multinuclear magnetic resonance analysis of two humic acid fractions from lowland rice soils.

To determine the effects of intensive cropping of tropical lowland rice (Oryza sativa L.) and the associated long-term soil submergence on chemical properties of soil organic matter, we used solid-state 13C and 15N and solution 31P nuclear magnetic resonance (NMR) spectroscopy to analyze the labile mobile humic acid (MHA) and the more recalcitrant calcium humate (CaHA) fractions extracted from a series of soils supporting several long-term field experiments in the Philippines. The soils varied mainly in degree of submergence and cropping intensity, ranging from a rainfed rice field without soil submergence to irrigated double- and triple-cropped fields in which soil remains submerged almost all year long. As reported previously, all analyses associated increasing intensity of rice cropping with larger proportions of less humified material in the MHA and CaHA, such as diester phosphorus (P), amide nitrogen (N), and phenolic carbon (C). We established significant correlations between proportions of various spectral areas as well as between some spectral areas and other humic acid (HA) properties such as visible light absorption and free radical concentration (positive indices of humification) and hydrogen (H) concentration (negative index of humification). For example, spectral proportions of heterocyclic N were positively, and proportions of amide N and phenolic C negatively, correlated with visible light absorption and free radical concentration, and each of these spectral proportions had an opposite sign when correlated with H concentration. The correlations of N-alkyl C proportions were the strongest with these properties and with other functional group proportions.

Stray field nuclear magnetic resonance of soil water: development of a new, large probe and preliminary results.

Development, characterization, and preliminary results of a recent technique capable of local measurements of pore-size distribution by a spatially resolved low resolution nuclear magnetic resonance (NMR) technique are described. Potential environmental uses include studying the change in pore-size distribution caused by surface compaction, which influences surface runoff, and obtaining information on the physical state of non-aqueous compounds in porous materials, which should aid the selection of appropriate soil remediation methods. Stray field (STRAFI) imaging is an NMR technique that allows distortion-free imaging of materials with short NMR relaxation times. The sample is placed in the strong axial fringe field gradient of a superconducting NMR magnet. We report on a new, unique, large 5-cm-diameter STRAFI probe, and its use for three preliminary test cases: water in ceramics of known pore size, paraffin wax and oil in sandstone rock, and water in soil at different matric potentials. The imaging is confined to one dimension with a spatial resolution of the order of 100 microm for protons. The optimum position for imaging occurs at 2.62 T and a gradient of 12.1 T/m. Water relaxation decay curves can be measured at any position in the 8-cm-long sample. These curves are decomposed into a series of terms each corresponding to a different pore size. Preliminary results show continuum fits to decay curves for a soil drained to three different matric potentials. Such information will be useful for interpreting water retention curves and will lead to understanding of the behavior of fluids in the vadose zone.

Phosphorus-31 solid-state nmr in high-field gradients: prospects for imaging bone using the long echo-train summation technique (LETS).

Stray-field techniques are reported for 31P studies of solids for a variety of compounds including bone, bone meal and calcium hydroxyapatite. Long Hahn echo trains produced by the application of many pulses were used as in the long echo-train summation technique. Double-resonance enhancements of 31P by use of both direct and indirect experiments were attempted on a sample of NH4PF6:31P[19F] double resonance produced, at most, a 26% increase in the initial level of the 31P echo signal.

STRAFI-NMR studies of water transport in soil.

1-D STRAFI (STRAy FIeld) imaging is used to study water distribution in a sandy loam. The matric potential of the soil can be varied during acquisition of 1-D profiles. Results at a range of potentials are presented showing both the equilibrium distribution and the evolution of the profile following an abrupt change in matric potential. The air breakthrough point and variations in draining behaviour due to differences in soil compaction are identified.

T1-weighting of Hahn echo-trains in the stray-field for deuterium: prospects for imaging using long echo-train summation.

Hahn-echoes have been observed in stray-field gradients of 37.5 T/m and 74.5 T/m for deuterium in heavy water in a number of forms: liquid, ice, and coordinated in solid copper sulfate and silica gel. In the diamagnetic samples, the application of many pulses (> 1000 or more) in long pulse-trains produced many sustained echoes. This lengthening of T(echo) is caused by T1-weighting of the components of the Hahn echo (even in the absence of spin-locking). In contrast, for the paramagnetic complex, the shortening of T1 greatly reduces T(echo) and only few echoes are obtained. A new protocol for stray field imaging is suggested when T1 is long: long echo-train summation (LETS). The observed magnetisation is much greater for heavy ice than for heavy water because the line-width, and hence the thickness of the excited slice, is greater.

Hahn-echoes from 14N in solids by the stray-field method: prospects for imaging using Long Echo-Train Summation.

We have observed Hahn-echoes resulting from Powles-Mansfield pulse-sequences for a variety of nitrogen containing solids which exhibit a range of 14N electric quadrupole coupling constants (Cq) from 0 to 4.9 MHz. Long echo-decays were frequently obtained which allowed the collection of many echoes in one echo-train, so that the method of Long Echo-Train Summation (LETS), could be used to accumulate signal. A one-dimensional STRAFI-profile is shown for solid ammonium nitrate. The spatial resolution is discussed as a function of Cq.

Stray-field imaging of quadrupolar nuclei of half integer spin in solids.

A report is presented on the observation of Hahn echoes from the following quadrupolar nuclei of half integer spin (I) in polycrystalline solids in the large static magnetic field gradient (37.5 T/m) which exists in the fringe field of a superconducting solenoid: 7Li, 23Na, 11B, 65Cu (I = 3/2); 27Al (I = 5/2); 51V, 59Co (I = 7/2); and 115In (I = 9/2). 23Na echo-trains from NaCl (with non-selective excitation) and from Na2SO4 (with selective excitation) are compared quantitatively for two different RF pulse sequences: 90x-(tau-90y-tau-echo-)n and 90x-(tau-90x-tau-echo-)n. The signals obtained from RF pulses corresponding to non-selective 90 degrees pulses were shown to be quantitative, whereas in the selective case smaller signals were obtained since only the central transition contributed. The loss of signal from this cause can be distinguished from small signals resulting from low density of nuclei by use of the second sequence. A 7Li image obtained from LiF in a cylindrical glass-vial is shown.

1H and 19F magnetic resonance imaging of solid paramagnetic compounds using large magnetic field gradients and Hahn echoes.

1H and 19F spatially resolved echo trains of a number of paramagnetic solids with apparent magnetic moments as high as 10 Bohr magnetons have been successfully obtained in the 58 T m-1 fringe-field gradient of a 9.4 T superconducting magnet by the use of Hahn echoes. Complexes studied include hydrates of Ni(II), Fe(III), Mn(II), and the anhydrous fluorides of Ti(III), Co(II) and Mn(II). Tris 6,6-7,7-8,8,8 heptafluoro-2,2-dimethyl-3,5octane dionato complexes of several lanthanides, Eu, Dy, Ho and Yb, have also been imaged. Additionally, 1H relaxation time measurements have been obtained at 28 MHz with conventional techniques, and several additional T2 values have been obtained in the stray field for both 1H and 19F.

A convenient method for calibration of the pulse-length in high field-gradients using Hahn echo-trains.

A new method is presented for calibration of the pulse-length in the case that the sample, which can be either a liquid or a solid, is placed in a high field-gradient such as it experiences in the stray-, fringe-field of a solenoid. The method employs a pulse-train, with a constant phase, of the form alpha X-tau-(alpha X-tau-echo-tau-)n. This produces Hahn echoes which have variable phase in the form of phase-alternations along the echo-train. For alpha = 90 degrees the relative phase-sequence in degrees is (0, 0, 180, 180), and we may characterise this by m = 2, since there are two signals of one phase in each group. For other cases with these phase angles (0 and 180) but different numbers, m, of signals with opposite phase, we have alpha = 180/m, where m is an integer.

Magnetic susceptibility effects in imaging: distortion-free images of plant tissue in soil.

Magnetic susceptibility effects (MSE) in NMR spectra are well known, and indeed the NMR technique has frequently been applied to measure magnetic susceptibility. In the case of imaging, MSE can lead to image distortion when the sample is heterogeneous. We have performed experiments on a soil sample (iron content approximately 2%) containing plant tissue which gave a NMR signal that was spread over about 15 kHz in the 1H spectrum. We present some results from a 128 x 128 x 128 3D 1H image (voxel size = 150 x 150 x 150 micron3) generated by the stray field imaging (STRAFI) technique in which the use of a 5 kG cm-1 magnetic flux density gradient reduced the magnetic susceptibility distortion to less than 10 microns.

Betaine metabolism in human neonates and developing rats.

Proton nuclear magnetic resonance spectroscopy has been used to demonstrate the presence of high concentrations of betaine (up to 0.75 mol/mol creatinine) in the urine of normal healthy human neonates. Betaine is not normally excreted in adults. Excretion of betaine from birth to 7 days old was monitored. The excretion of betaine in rats from 21 days after birth to 40-45 days old was also monitored. A peak in excretion in the rats of 1.5-3 mol/mol creatinine occurred between days 30-35. The presence of a high concentration of betaine in the urine is unlikely to be caused by a relative lack of betaine homocysteine methyl transferase activity compared with adults but may relate to the disposal of dietary choline during development.

The conformation of gramicidin A in dimethylsulphoxide solution. A full analysis of the one- and two-dimensional 1H, 13C, and 15N nuclear-magnetic-resonance spectra.

The combined application of one- and two-dimensional high-field NMR techniques has led to the first assignment of the 1H, 13C, and 15N spectra of the pentadecapeptide gramicidin A in dimethylsulphoxide solution. The 62.9-MHz and 100.6-MHz 13C spin-lattice relaxation times and 13C-[1H] NOE factors for the backbone alpha carbons have been analysed in the 'model-free' approach to give a single correlation time (tau m) for isotropic overall molecular motion and an order parameter and internal correlation time for each C alpha H group in the backbone. The relatively high and constant values for the order parameter along the backbone indicate a degree of ordering of the structure, while the internal correlation times show that internal motions are progressively more rapid towards the N terminus. The average values of the vicinal HNC alpha H couplings are 7.4 Hz and 8.4 Hz respectively for the alternate L- and D-amino acid residues. The values are not consistent with either a ribbon conformation for the backbone or a right-handed beta 6.3 helix; they are consistent with the model proposed by Glickson et al. [Glickson, J. D., Mayers, D. F., Settine, J. M. & Urry, D. W. (1972) Biochemistry 11, 477-486] in which there is a rapid conformational order in equilibrium disorder equilibrium, the ordered structure being the left-handed beta 6.3 helix and the disordered state having local random-coil character.

Qualitative aspects of hydrogen-deuterium exchange in the 1H, 13C, and 15N nuclear magnetic resonance spectra of viomycin in aqueous solution.

The 1H, 13C, and 15N high field nuclear magnetic resonance spectra of the cyclic peptide viomycin have been fully assigned using homo- and heteronuclear double resonance experiments and pH effects. In addition it is shown how the two- and three-bond H-D isotope effects upon carbonyl resonances may assist in their assignment. The resistance to exchange with solvent water of the amide proton involved in the transannular hydrogen bond is observed directly in the 1H spectra, via the isotope effect on a carbonyl resonance in the 13C spectra, and via the one-bond 1H couppling in the 15N spectra.

Theory and practice for studies for peptides by 15N nuclear magnetic resonance at natural abundance: gramicidin S.

The low-abundance isotope of nitrogen, 15N, is now accessible to study by the latest high resolution nuclear magnetic resonance techniques. Structure and motion in polypeptides of moderate size may now be usefully investigated in this way.

Nuclear magnetic resonance and infrared studies of N15-substituted trimethylsilyl-, -germyl-, and -stannylanilines. A Test of (p->d)-pi interactions in group IV nitrogen bonds.

Infrared and proto n magnetic resonance spectra of the aniline-N14 and -N15 isotopomers and their N-trimethylsilyl, -germyl, and -stannyl derivatives have been measured and compared. Nmr assignments have been checked by comparisons of proton spectra at 60 and 100 Mcps; by proton-N15 heteronuclear experiments at 9400 G; and by observation of N15 satellites. The one-bond "5-H couplings can be interpreted (on the assumption of a dominant Fermi spin-spin interaction, small radial variations for nitrogen wave functions, and equal distributions of s character in the sigma bonds) in terms of pyramidal arrangements of bonds at nitrogen. The relation of the stereochemical situation at nitrogen to the question of (p-->d)-pi or (p-->p)-pi bonding involving nitrogen is discussed.