Quantitative differences in evaluating soil humic substances by liquid- and solid-state C-NMR spectroscopy

We compared the quantitative responses of liquid-state (LS) and solid-state (CPMAS) ¹³C-NMR spectroscopy of four different soil humic substances. The intensities of signals for the alkyl carbons (0–40 ppm) were significantly larger in CPMAS than in LS spectra. This difference is in agreement with the pseudo-micellar model of the conformational nature of humic substances. By this view, the hydrophobic interactions holding together the heterogeneous molecules of humic micelles inhibit the molecular motions of the alkyl carbons, thereby enhancing the spin-lattice relaxation times and consequently lowering the sensitivity of liquid-state NMR. Conversely, regardless of their position in the humic conformation, a better estimation of the number of alkyl carbons can be obtained by CPMAS-NMR because of the cross-polarization of hydrogen nuclei in CH₂ and CH₃ groups. The intensity of the 40–110 ppm region is also slightly lower in LS than in CPMAS-NMR spectra, despite the hydrophilicity of the oxidized and peptidic carbons resonating in this chemical shift interval. Their molecular motion may also be reduced by either the formation of intra- and inter-molecular hydrogen bondings due to poorly acidic hydroxyl groups of saccharides, or the degree of conformational rigidity that a pseudo-micellar arrangement confers even to hydrophilic domains. The higher content of aromatic carbons (110–160 ppm) found in the LS spectra was attributed partly to the high degree of substitution of the aromatic ring that slows down cross-polarization in CPMAS experiments and partly to the relative overestimation of this region by LS-NMR due to a lack of signal in the aliphatic interval. The slightly lower content of carboxyl carbons estimated in CPMAS spectra as compared to LS spectra was also attributed to slow cross-polarization. This work shows that the combined use of both NMR techniques is profitable in conformational analysis of humic substances and of dissolved organic matter in general.     

13C NMR studies of organic matter in whole soils: I. Quantitation possibilities

A detailed discussion of the quantitative nature of ¹³C CPMAS NMR spectra as applied to solid samples, such as soil, is given. In particular, the influence of the cross-polarization (CP) time constant (T_CH), the relaxation time constant of protons in the rotating frame (T_1pH) and the contact time (t_c) in the CPMAS experiment are considered. Three distinct quantitation regimes are numerically identified according to sample parameters t_CH and T_1PH > and the experimental choice of t_c: (i) quantitation obtainable from a single CPMAS spectrum; (ii) quantitation obtainable from a series of CPMAS spectra; and (iii) quantitation not possible using CPMAS. Strategies for the measurement of sample parameters T_CH and Ti_pH are reviewed. When quantitation is not possible using CPMAS it is necessary to regress to the direct polarization (DP) of ¹³C nuclei. The sensitivity problems of DPMAS are discussed, as too are general factors that affect the quantitation of ¹³C data such as spinning sidebands. More specifically in relation to soil samples, the effects on quantitation arising from the presence of paramagnetics and the actual methods for the measurement of signal intensities are covered.     

Effects of mineral surface iron on the CPMAS 13C-NMR spectroscopic detection of organic matter from soil fractions in an agricultural topsoil with different amendments

The decrease of NMR visibility of the C signal in soil samples due to the association between organic carbon (OC) and the topsoil mineral surface was investigated. CPMAS ¹³C‐NMR spectra were obtained for soil particle‐size fractions (< 2 μm, 2–20 μm, > 20 μm) and bulk soils from an agricultural topsoil (Chernozem) that had received three different amendments (no fertilization, mineral fertilization (NPK), mineral (NPK) and organic (cattle manure) fertilizations) at Bad Lauchstädt, Germany. The soil organic carbon content of the three soils depended on the degree of soil fertilization. There was no constant relationship between the total NMR signal intensity and the total amount of organic carbon (TOC) for all size fractions. Indeed, a key role played in the C signal intensity by the paramagnetic ferric ion from the clay content in soil fractions and bulk soils was confirmed. Thus, we describe the variations of C signal intensity by taking into account the distribution of clay‐associated OC and non‐associated OC pools. Depending on the amendment, the C signal visibility was weakened by a factor of 2–4 for the clay‐associated OC. This estimation was rendered possible by combining mineral specific surface area (SSA) measurements with the N₂ gas adsorption method (BET method) and determination of TOC and iron concentrations. This approach contributes to the quantitative evaluation of the CPMAS ¹³C‐NMR detection.     

Analysis of protein structures and interactions in complex food by near-infrared spectroscopy. 2. Hydrated gluten

Hydrated gluten, treated with various salts, was analyzed by near-infrared (NIR) spectroscopy to assess the ability of this method to reveal protein structure and interaction changes in perturbed food systems. The spectra were pretreated with second-derivative transformation and extended multiplicative signal correction for improving the band resolution and removing physical and quantitative spectral variations. Principal component analysis of the preprocessed spectra showed spectral effects that depended on salt type and concentration. Although both gluten texture and the NIR spectra were little influenced by treatment with salt solutions of low concentrations (0.1-0.2 M), they were significantly and diversely affected by treatment with 1.0 M salt solutions. Compared to hydration in water, hydration in 1.0 M sulfate salts caused spectral effects similar to a drying-out effect, which could be explained by salting-out.     

Quantitative aspects of solid-state C-NMR spectra of humic substances from soils of volcanic systems

Cross-Polarisation Magic Angle Spinning Carbon-13 Nuclear Magnetic Resonance spectroscopy (CPMAS ¹³C-NMR) represents one of the most powerful tools to investigate soil organic matter (SOM) mainly because of its inherent capacity to provide a semi-quantitative evaluation of carbon distribution. A critical parameter during acquisition of CPMAS ¹³C-NMR spectra is the contact time required to obtain the cross-polarisation between proton and carbon nuclei. The procedure to evaluate the best contact time for the acquisition of a quantitative CPMAS ¹³C-NMR spectrum is to perform Variable Contact Time (VCT) experiments. In this work the structural features of a number of purified humic substances from Italian and Costarican volcanic soils were investigated by CPMAS ¹³C-NMR spectroscopy after having performed preliminary VCT experiments. The VCT experiments showed that the average contact times vary according to the origin and chemical structure of the humic material. The optimal contact times (OCT) for nine humic samples were between 250 and 800 μs These values were different from the time of 1000 μs that is commonly applied as the best average contact time for humic materials. Moreover, by comparing the NMR data to those obtained by elemental analysis (C/H ratio), it appeared that the efficiency of the cross-polarisation between protons and carbons, and hence the contact time, is affected not only by the number of protons, but also by their distribution over the molecules. The evaluation of errors in quantitative estimation of the different carbons revealed that the use of OCT generally reduced by half the loss of signals occurring when the average contact time of 1000 μs is used in CPMAS ¹³C-NMR spectra of humic substances.     

Multivariate analysis of CPMAS 13C-NMR spectra of soils and humic matter as a tool to evaluate organic carbon quality in natural systems

A series of humic and fulvic acids isolated from different sources, size‐fractions separated from a humic acid, and three soils of different origin were subjected to CPMAS ¹³C‐NMR spectroscopy to obtain the distribution of their carbon contents. The relative areas of chemical shift regions in NMR spectra were used to apply a principal component analysis (PCA) to the three sets of samples. The multivariate analysis was successful in efficiently differentiating samples on the basis of the quality of their organic carbon content. The PC biplots based on two principal components distinguished objectively among samples as accurately as it was possible to do by subjective qualitative evaluation of the original spectra. In the case of the soils, a discriminant analysis (DA) was applied to build a classification model that allowed the validation of the three soils according to their origin. Percentage of validation in the classification model is expected to increase when a large number of NMR spectra are accumulated and/or the concentration of organic carbon in samples is enhanced. The multivariate analyses described are likely to become a useful tool to increase the importance of CPMAS ¹³C‐NMR spectra in the appraisal of natural organic matter variations in heterogeneous natural systems.     

Organic N forms of a subtropical Acrisol under no-till cropping systems as assessed by acid hydrolysis and solid-state NMR spectroscopy

This study was conducted to investigate the influence of land-use systems (grassland and cropland) and of long-term no-till cropping systems [bare soil, oat/maize (O/M), pigeon pea+maize (P+M)] on the composition of organic N forms in a subtropical Acrisol. Soil samples collected from the 0- to 2.5-cm layer in the study area (Eldorado do Sul RS, Brazil) were submitted to acid hydrolysis and cross-polarization magic angle spinning (CPMAS) ¹⁵N and ¹³C nuclear magnetic resonance (NMR) spectroscopies. The legume-based cropping system P+M contained the highest contents of non-hydrolysable C and N, hydrolysable C and N, amino acid N and hydrolysed unknown N. The relative proportion of non-hydrolysable N was higher in bare soil (30.0%) and decreased incrementally in other treatments based on the total C and N contents. The amino acid N corresponded to an average of 37.2% of total N, and was not affected by land use and no-till cropping systems. The non-hydrolysable residue contained lower O-alkyl and higher aromatic C concentrations, as revealed by CPMAS ¹³C NMR spectroscopy, and higher C:N ratio than the bulk soil. No differences in the bulk soil organic matter composition could be detected among treatments, according to CPMAS ¹³C and ¹⁵N NMR spectra. In the non-hydrolysable fraction, grassland showed a lower concentration of aromatic and a higher concentration of alkyl C than other treatments. From CPMAS ¹⁵N NMR spectra, it could be concluded that amide N from peptide structures are the main organic N constituent. Amide structures are possibly protected through encapsulation into hydrophobic sites of organic matter and through organomineral interaction.     

Fast field cycling NMR relaxometry characterization of biochars obtained from an industrial thermochemical process

Purpose

Biochar has unique properties which make it a powerful tool to increase soil fertility and to contribute to the decrease of the amount of atmospheric carbon dioxide through the mechanisms of C sequestration in soils. Chemical and physical biochar characteristics depend upon the technique used for its production and the biomass nature. For this reason, biochar characterization is very important in order to address its use either for agricultural or environmental purposes.

Materials and methods

Three different biochars obtained from an industrial gasification process were selected in order to establish their chemical and physical peculiarities for a possible use in agronomical practices. They were obtained by charring residues from the wine-making industry (marc) and from poplar and conifer forests. Routine analyses such as pH measurements, elemental composition, and ash and metal contents were performed together with the evaluation of the cross-polarization magic angle spinning (CPMAS) ¹³C NMR spectra of all the biochar samples. Finally, relaxometry properties of water-saturated biochars were retrieved in order to obtain information on pore size distribution.

Results and discussion

All the biochars revealed basic pH values due to their large content of alkaline metals. The quality of CPMAS ¹³C NMR spectra, which showed the typical signal pattern for charred systems, was not affected by the presence of paramagnetic centers. Although paramagnetism was negligible for the acquisition of solid state spectra, it was effective in some of the relaxometry experiments. For this reason, no useful information could be retrieved about water dynamics in marc char. Conversely, both relaxograms and nuclear magnetic resonance dispersion profiles of poplar and conifer chars indicated that poplar char is richer in small-sized pores, while larger pores appear to be characteristic for the conifer char.

Conclusions

This study showed the potential of relaxometry in revealing chemical?Cphysical information on industrially produced biochar. This knowledge is of paramount importance to properly direct biochar agronomical uses.     

Chemical properties of humic matter as related to induction of plant lateral roots

A series of humic matter samples isolated from a soil sequence, different oxisols, size‐fractionated from a vermicompost humic acid and subjected to chemical modifications, were characterized by CPMAS ¹³C‐NMR spectroscopy. The relative signal areas in chemical shift regions of NMR spectra of the four sets of samples were analysed by principal component analysis (PCA). Hierarchical cluster analysis (HCA) was applied to build a classification model, which allowed the recognition of humic matter according to its origin. The relationship between carbon species and biological activity of humic acids, as promoters of lateral root emergence, was obtained by applying PLS multivariate analysis. This showed that lateral root emergence was mostly related to NMR parameters such as the hydrophobicity index (HB/HI) and the 40–110 and 160–200 ppm chemical shift regions (hydrophilic carbon HI), while the content of hydrophobic (HB) carbon in humic samples was negatively correlated with induction of lateral root hair. Our results represent a step further in the structure‐bioactivity relationship of natural humic substances and confirm their role as plant root growth promoters.     

Immobilization of 15N in forest litter studied by 15N CPMAS NMR spectroscopy

Solutions labelled with ¹⁵N were applied as (¹⁵NH₄)₂SO₄ or K¹⁵NO₃ to isolated microplots in the floor of mountain beech forest (Nothofagus solandri var. cliffortioides) and incubated for 135 days under field conditions of moisture and temperature. Solid state ¹⁵N CPMAS NMR spectra of the forest litter layer showed that more than 80% of the total signal intensity was attributable to the secondary amide-peptide peak. The degree of ¹⁵N enrichment or form of N did not alter the relative intensity of signals attributable to ¹⁵N in peptides, nucleic acids and aliphatic amine groups (amino sugars and free NH₂ on amino acids). Combinations of ¹³C and ¹⁵N-NMR spectra, edited by a process that exploited differences in proton spin properties between distinct categories of organic matter, indicated incorporation of ¹⁵N in humified organic matter rather than partly degraded plant material. This application demonstrated that solid state ¹⁵N CPMAS NMR has potential for use in studies of N immobilization under field conditions and with materials containing little N and small ¹⁵N enrichment.     

Temporal changes in distribution and composition of N from labeled fertilizer in soil organic matter fractions

Variations in the amount and composition of immobilized nitrogen (N) in major soil organic matter fractions were investigated in a 730-day soil incubation experiment using ¹⁵N-labeled urea and ¹⁵N nuclear magnetic resonance spectroscopy with the cross polarization/magic angle spinning (¹⁵N CPMAS NMR) method. After 730 days, 24.7% of the applied N was recovered from the soil as organic N. The urea-derived N recovered from humic acids and humin decreased from 11.2 and 33.8% of the applied amount after 14 days to 1.6 and 20.4% after 730 days, respectively. When these values were corrected for the microbial biomass (MB) N, they ranged from 9.0 to 1.2% and 28 to 18%, respectively. The proportion of urea-derived N recovered from fulvic acids was low, ranging between 0.4 and 5.8% (with MB N) or 5.6% (without MB N) of the applied amount, whereas that from water-soluble nonhumic substances (WS-NHS; NHS in the fulvic acid fraction) remained high, 28–33% of the applied amount after correction for the contribution of MB N up to day 365, and decreased to 0.9% thereafter. The ¹⁵N CPMAS NMR spectra of humic acids, fulvic acids, and humin showed the largest signal at −254 to −264 ppm, corresponding to peptide/amide N. The proportions of heterocyclic, peptide/amide, guanidine/aniline, and free amino N in the urea-derived humic acid N were 3–7, 83–90, 5–7, and 2–4%, respectively. More than 80% loss of the urea-derived humic acid N did not markedly alter their composition. No time-dependent variations were also observed for the proportions of respective N functional groups in humin N, which were 3–5, 71–78, 12–17, and 6–10% in the same order as above. These results suggest the greater importance of physical stability than structural variation for the initial accumulation of organic N in soil.     

Catechin transformation as influenced by aluminum

Polyphenols (catechins) are vital biomolecules in tea plants (Camellia sinensis), which are well-known as typical Al accumulators. However, the interaction between Al and catechin remains obscured. The objective of the present study was to investigate the effect of Al on the transformation of (+)-catechin. Solutions with OH/Al molar ratios of 2.5 (pH 5.5) and 3.0 (pH 7.0) prepared at Al/catechin molar ratios (R) of 0, 0.2, 0.4, 0.6, 0.8, and 1.0 were aged for 7 and 30 days, respectively. The precipitates were collected and examined by wet chemistry, X-ray diffraction, transmission electron microscopy, electron spin resonance (ESR), cross-polarization magic angle (CPMAS) 13C nuclear magnetic resonance (13C NMR) analyses, and Fourier transformation infrared absorption spectrometry (FT-IR). The weight of the precipitates increased with increasing Al/catechin molar ratios and with prolonged aging. The molar ratios of Al/catechin in the precipitates increased with increasing initial Al/catechin molar ratios and were close to the initial solution Al/catechin molar ratios. The chemical analysis and spectroscopic studies indicated that Al was bonded with catechin, forming a 1:1 type complex. The reaction of crystalline catechin with Al resulted in the formation of X-ray noncrystalline precipitates. The solid-state CPMAS 13C NMR spectra of the precipitates show the change in chemical shifts of catechin as a result of catechin complexation with Al. The FT-IR spectra of the Al-catechin precipitates also show the loss of absorption bands of several functional groups compared with catechin. The FT-IR data substantiate this reasoning. The ESR spectra of the precipitates show a single symmetrical line devoid of any fine splitting, indicating the presence of free radicals of semiquinones, which are commonly present in humified materials.     

Effects of Temperature on Sorption of Water by Wheat Gluten Determined Using Deuterium Nuclear Magnetic Resonance

The effects of lipids and residual starch components of wheat flour gluten on gluten hydration properties were investigated using nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR) techniques. Whole or native, lipid-free, starch-free, and lipid- and starch-free gluten samples were prepared from wheat (Triticum aestivum) cv. Mercia. ²H NMR relaxation on gluten samples hydrated with deuterium oxide (D₂O) was measured over a 278–363 K temperature range. FTIR spectra were recorded in dry and fully hydrated material. Transverse relaxation (T₂) results indicated that all four gluten samples were hydrophilic in nature. There was little difference in relaxation behavior of whole and lipid-free gluten samples. T₂ values and populations of the relaxation components were very similar in each. The FTIR spectra of both samples showed an increase in extended β-sheet secondary structures on hydration. These results suggest that lipid binding in gluten, if it occurs, has little effect on wheat gluten properties. Adding starch to the gluten matrix results in an increase in water sorption on heating that may be attributed to the effects of starch gelation. However, the whole water uptake of the gluten cannot be accounted for by the contribution of the residual starch, as estimated by the effects of added starch. Extraction of residual starch required solubilization of the protein, including breaking of the disulfide bonds. This process altered the gluten structure and properties. Light microscope investigation showed that glutens with residual starch extracted were unable to form fibrillar strands on hydration. NMR and FTIR results showed greater water sorption in both samples with extracted starch than in the unextracted samples.     

Long‐term effects of liming on microbial biomass and activity and soil organic matter quality (13C CPMAS NMR) in organic horizons of Norway spruce forests in Southern Germany

Long‐term effects of liming on microbial biomass and activity and soil organic matter (SOM) were investigated in samples from organic horizons (Of/Oh) in spruce forests at Adenau, Höglwald, Idar‐Oberstein, and Schluchsee (Southern Germany) where plots have been manually treated 7 to 13 years ago with dolomitic limestone. At all sites, pH values were markedly increased after liming. The contents of C and N in the organic horizons of the limed plots appeared to be lower with the greatest decrease at Höglwald (Dystric Luvisol) where liming has affected the soil properties for the longest time of all sites. Catalase activity was promoted after liming at Adenau (Cambic Podzol). This was also the case for the Dystric Luvisol where liming resulted also in higher basal respiration. Biomass‐C was higher in samples from the limed plot at Idar‐Oberstein (Dystric Cambisol). The ¹³C CPMAS NMR spectra of organic horizons from the control plots indicate no differences in the gross carbon composition of SOM. Furthermore, spectra from the limed Cambic Podzol, Dystric Cambisol, and Haplic Podzol (Schluchsee) were remarkably similar. However, for the Dystric Luvisol, the lime‐induced promotion of microbial activity resulted in lower O‐alkyl‐C intensity. The observed patterns of microbial biomass and activity were site‐dependent rather than a result of liming. Obviously liming had only small long‐term effects on the humus quality in the organic horizons, as far as detectable by CPMAS NMR spectroscopy. More sensitive techniques like pyrolysis‐GC/MS should be applied to analyze differences in C composition.     

The immobilization of nitrogen by straw decomposing in soil

Immobilization of nitrogen (N) in decomposing straw varies between soils, and the objective of this study was to identify the mechanisms responsible. Internode segments of wheat straw were incubated in Denmark and in Scotland in arable soils fertilized with NH₄NO₃, labelled with ¹⁵N, for periods up to 1 year. Straw was recovered from the soils periodically and analysed for microbial biomass and different forms of N using chemical methods and CPMAS ¹⁵N NMR spectroscopy. The total N content of the straw increased, as long as the soil was not too wet, such that there was overall immobilization. This was accompanied by a rapid increase in the content of amino acid N and to a lesser extent of glucosamine N and a concomitant decrease in the carbohydrate content of the straw. Using direct and plate counts for bacterial and ergosterol content for fungal estimation, we found that fungal biomass was much greater than that of bacteria. This correlated with the forms of N in the straw as determined by CPMAS ¹⁵N NMR, which showed spectra that were more typical of fungi than of bacteria. It seems that immobilization of N is primarily caused by fungi as they decompose the straw.     

Temporal changes in humic acids in cultivated soils with continuous manure application

Abstract

Although the application of manure to upland fields is believed to induce changes in the quality of humic substances in soil as well as the quantity, the direction and extent of these changes have not been elucidated. To understand temporal variations in humic acids, periodically collected soil samples from two fields, a Typic Hapludult (Togo) and a Pachic Melanudand (Kuriyagawa), with cattle manure and chemical fertilizer (CF) were examined. The content and degree of humification (darkening) of the humic acids were distinctly greater in Kuriyagawa than in Togo soil. Corresponding to the difference in the degree of humification, molecular size distribution, elemental composition, infrared (IR) spectra, and ¹³C cross polarization/magic angle spinning nuclear magnetic resonance (CPMAS NMR) spectra of humic acids differed between the two soils. Manure application at 40 Mg ha^?1 year^?1 for 16 years (Togo) and at 80 or 160 Mg ha^?1 year^?1 for 19 years (Kuriyagawa) resulted in greater humic acid content compared with plots with CF only because of its increase in the manured plots and/or decrease in the CF plots. Manure application at an extremely high rate (160 Mg ha^?1 year^?1) resulted in higher H content and greater signal intensities of alkyl C, O-alkyl C and amide C=O in the ¹³C CPMAS NMR and/or IR spectra. Although humic acids with larger molecule sizes increased in all the manured plots, differences between the humic acids from the plots with and without manure applied at practical levels in the elemental and spectroscopic analyses were small or scarce. These results were considered to be because of the similarity between the indigenous soil humic acids and the manure-derived ones in Togo soil (a low degree of humification) and because of the abundance of highly-humified humic acids in Kuriyagawa soil.     

Sugar-casein interaction in deuterated solutions of bovine and caprine casein as determined by oxygen-17 and carbon-13 nuclear magnetic resonance: a case of preferential interactions

17O NMR spectroscopy and (13)C NMR spectroscopy have been used to study the mechanism of interaction of sugars with bovine and caprine caseins in D(2)O. The (17)O NMR relaxation results showed in all cases an increase in water of hydration, as a result of added sugar; this was predominantly associated with "trapped" water in the caseins. Analysis of the vir al coefficients, obtained from the (17)O relaxation data, suggested that preferential interactions occur in the sugar-protein solutions. This could be the result of either sugar binding or a solute-solute thermodynamic effect, preferential hydration. The addition of sugars to deuterated solutions of bovine casein and caprine casein high in alpha(s1)-casein had little or no effect on either line width or chemical shifts of the (13)C NMR spectra of these milk proteins. (13)C NMR studies of sucrose, at various concentrations (100-300 mM) in the presence of caprine casein high in alpha(s1)-casein, showed no changes in either chemical shifts or T(1) values. This indicates that the sugar molecules tumble isotropically and therefore neither bind to the protein nor affect viscosity in the protein-sugar studies. All of these data suggest that the preferential exclusion of the sugar from the domain of the caseins results in preferential hydration of the caseins.     

1H nuclear magnetic resonance relaxometry study of water state in milk protein mixtures

1H NMR signal was used to characterize highly hydrated milk protein dispersions (3-20% dry matter) with various micellar casein concentrations (3-15%), whey protein concentrations (0-3%), lactose concentrations (0-7.5%), CaCl(2) concentrations (0-2 mM), and pH (6.2-6.6). The results showed the predominant effect of micellar casein concentration on water state and were consistent with the three-site relaxation model in the absence of lactose. The relaxation rates observed for these dispersions were explained by the free water relaxation rate, the hydration water relaxation rate, and the exchangeable proton relaxation rate. Hydration water was found to be mainly influenced by casein micelle concentration and structure. The variations in hydration with pH were consistent with those observed for classical measurement of voluminosity observed at this range of pH. The effects of lactose and whey protein content are discussed.     

Impacts of Soil Management Practices on the Organic Matter Structure - Investigations by CPMAS 13CNMR-Spectroscopy

Humus properties of 42 soil samples or humic extracts of Ap horizons of Cambisols and Luvisols, from different field plots, kept for a long time under continuous management conditions, have been studied by quantitative CPMAS ¹³C NMR-spectroscopy. The shift range of the spectra were divided into 8 regions, with carbon absorptions from 0–45 (aliphatics), 45–60 (OCH₃), 60–80 (CO? CN), 80–110 (anomeric carbons), 110–140 (aryl—C), 140–160 (O-aryl-C) and 160–210 (COOH? C?O) ppm, respectively. Although soil samples and humic extracts were obtained from plots from different locations and sometimes widely differing organic carbon contents, variances in the relative absorptions of the selected ranges were rather small. Several absorptions were significantly correlated among each other or with soil carbon and microbial biomass contents. These correlations were discussed with the applied management and with other results about humus formation and properties. CPMAS ¹³C NMR-spectroscopy allows a reliable and comprehensive characterization of soil organic matter from soils without previous fractionation.     

Two‐Dimensional Vibration Spectroscopy of Rice Quality and Cooking

Rice samples were taken from a study of rice milling properties that affect quality. The spectra of milled and cooked samples were taken in the near‐infrared, mid‐infrared, and Raman region. These spectra, two regions at a time, were regressed by a two‐dimensional technique to develop contour maps that indicated the correlation of two spectral regions. These relationships demonstrate that it is possible to recognize the hydration effects caused by gelatinization (cooked samples vs. milled rice). Three water (O‐H stretch) spectral bands (960, 1445, 1,930 nm) in the near‐infrared (NIR) show marked differences between milled and cooked rice. The difference spectra indicated that there were additional phenomena occurring besides the addition of water. These differences are apparent in both C‐O‐H and N‐H bands, which indicate that water is interacting with both starch and protein. The two‐dimensional technique developed in this laboratory was used to get a better interpretation of what occurs during cooking. The Raman spectrum, which is relatively insensitive to water (O‐H stretch), revealed only changes in protein that could be associated with denaturization.