The effect of water content on solid-state 13C NMR quantitation and relaxation rates of soil organic matter

Two hydrofluoric acid‐treated soils were prepared with water contents ranging up to 22% by exposing them to a range of atmospheric humidities. There was no effect of water content on the chemical shift distribution of nuclear magnetic resonance (NMR) signal in ¹³C cross‐polarization (CP) NMR spectra. The sensitivity of the ¹³C CP NMR spectra decreased slightly with increasing water content. Much of this decrease could be attributed to decreases in T_1ρH relaxation rates, caused by enhanced molecular mobility of the organic matter in the presence of absorbed water. Rates of T_1H relaxation were very sensitive to water content, and average T_1H relaxation rates decreased four‐ to five‐fold from the smallest to the largest water content. Rates of T_1H relaxation were non‐uniform, and were better modelled by two‐T_1H component fits than one‐T_1H component fits. The ratio of rapidly to slowly relaxing components increased with increasing water content. Proton spin relaxation editing (PSRE) subspectra revealed substantial changes in the nature of these two components with increasing water content. These results indicate the presence of an organic matter component that is very sensitive to water content, transforming from slowly relaxing at a small water content to rapidly relaxing at a greater water content. This component was shown to be rich in O–alkyl and carbonyl C, and may be hemicellulosic root exudates and microbial mucilages. The slowly relaxing PSRE component was a mixture of ligno‐cellulose and alkyl biopolymers, whereas the rapidly relaxing component was primarily charcoal for one of the soils, and was reminiscent of dissolved organic carbon for the other soil. These findings show that care must be taken in controlling water contents when using PSRE to study organic matter.     

The influence of vegetational origin and degree of humification of organic soils on their chemical composition, determined by solid-state 13C NMR

Organic soil samples with different vegetational background and others with variation in the degree of humification, were investigated with solid-state ¹³C NMR. This work indicates that the vegetational origin and degree of humification of the organic matter appear to influence the distribution of functional groups in organic soils considerably, but one year of decomposition under controlled laboratory conditions gave only small changes in the chemical composition.     

Quantitative solid-state 13C NMR spectroscopy of organic matter fractions in lowland rice soils

Spin counting on solid‐state ¹³C cross‐polarization (CP) nuclear magnetic resonance (NMR) spectra of two humic fractions isolated from tropical lowland soils showed that only 32–81% of potential ¹³C NMR signal was detected. The observability of ¹³C NMR signal (C_obs) was higher in the mobile humic acid (MHA) than in the calcium humate (CaHA) fraction, and increased with increasing intensity of irrigated rice cropping. NMR observability appeared to be related to the nature of the organic carbon, with phenol‐ and methoxyl‐rich samples having the higher values of C_obs. The Bloch decay (BD) technique provided more quantitatively reliable ¹³C NMR spectra, as evidenced by values of C_obs in the range 91–100% for seven of the eight humic fractions studied. The BD spectra contained considerably more aryl and carbonyl signal, and less O–alkyl and alkyl signal, with the greatest differences between CP and BD spectra observed for the samples with low C_obs(CP). The causes of low CP observability were investigated using the spectral editing technique RESTORE ( RE storation of S pectra via T _CH and T O ne R ho (T_1ρH) E diting). Rapid T_1ρH relaxation was found to be primarily responsible for the under‐representation of carbonyl carbon, whereas inefficient cross‐polarization was primarily responsible for the under‐representation of aryl carbon in CP spectra. Proton NMR relaxation rates T_1H and T_1ρH were found to correlate with other NMR properties and also with cropping management. Non‐uniform rates of T_1H relaxation in two of the CaHA fractions enabled the generation of proton spin relaxation editing subspectra.     

The dynamics of organic matter in rock fragments in soil investigated by 14C dating and measurements of 13C

Rock fragments in soil can contain significant amounts of organic carbon. We investigated the nature and dynamics of organic matter in rock fragments in the upper horizons of a forest soil derived from sandstone and compared them with the fine earth fraction (<2 mm). The organic C content and its distribution among humic, humin and non‐humic fractions, as well as the isotopic signatures (Δ¹⁴C and δ¹³C) of organic carbon and of CO₂ produced during incubation of samples, all show that altered rock fragments contain a dynamic component of the carbon cycle. Rock fragments, especially the highly altered ones, contributed 4.5% to the total organic C content in the soil. The bulk organic matter in both fine earth and highly altered rock fragments in the A1 horizon contained significant amounts of recent C (bomb ¹⁴C), indicating that most of this C is cycled quickly in both fractions. In the A horizons, the mean residence times of humic substances from highly altered rock fragments were shorter than those of the humic substances isolated in the fine earth. Values of Δ¹⁴C of the CO₂ produced during basal respiration confirmed the heterogeneity, complexity and dynamic nature of the organic matter of these rock fragments. The weak ¹⁴C signatures of humic substances from the slightly altered rock fragments confirmed the importance of weathering in establishing and improving the interactions between rock fragments and surrounding soil. The progressive enrichment in ¹³C from components with high‐¹⁴C (more recent) to low‐¹⁴C (older) indicated that biological activity occurred in both the fine and the coarse fractions. Hence the microflora utilizes energy sources contained in all the soil compartments, and rock fragments are chemically and biologically active in soil, where they form a continuum with the fine earth.     

Characterization of recently 14C pulse-labelled carbon from roots by fractionation of soil organic matter

The inability of physical and chemical techniques to separate soil organic matter into fractions that have distinct turnover rates has hampered our understanding of carbon (C) and nutrient dynamics in soil. A series of soil organic matter fractionation techniques (chemical and physical) were evaluated for their ability to distinguish a potentially labile C pool, that is ‘recent’ root and root‐derived soil C. ‘Recent’ root and root‐derived C was operationally defined as root and soil C labelled by ¹⁴CO₂ pulse labelling of rye grass–clover pasture growing on undisturbed cores of soil. Most (50–94%) of total soil + root ¹⁴C activity was recovered in roots. Sequential extraction of the soil + roots with resin, 0.1 m NaOH and 1 m NaOH allocated ‘recent’ soil + root ¹⁴C to all fractions including the alkali‐insoluble residual fraction. Approximately 50% was measured in the alkali‐insoluble residue but specific activity was greater in the resin and 1 m NaOH fractions. Hot 0.5 m H₂SO₄ hydrolysed 80% of the ¹⁴C in the alkali‐insoluble residue of soil + roots but this diminished specific activity by recovering much non‐¹⁴C organic matter. Pre‐alkali extraction treatment with 30% H₂O₂ and post‐alkali treatment extractions with hot 1 m HNO₃ removed organic matter with a large ¹⁴C specific activity from the alkali‐insoluble residue. Density separation failed to isolate a significant pool of ‘recent’ root‐derived ¹⁴C. The density separation of ¹⁴C‐labelled roots, and roots remixed with non‐radioactive soil, showed that the adhesion of soil particles to young ¹⁴C‐labelled roots was the likely cause of the greater proportion of ¹⁴C in the heavy fraction. Simple chemical or density fractionations of C appear unsuitable for characterizing ‘recent’ root‐derived C into fractions that can be designated labile C (short turnover time).     

Spin accounting and RESTORE – two new methods to improve quantitation in solid-state 13C NMR analysis of soil organic matter

Rapid T_1ρH relaxation and inefficient cross‐polarization have long been known to affect quantitation in solid‐state ¹³C cross‐polarization (CP) NMR spectra of soil organic matter. We have developed two new techniques to overcome these problems. The first, spin accounting, enables accurate gauging of how quantitative a spectrum is likely to be. The result is expressed as the percentage of potential NMR signal that can be accounted for (C_obs). Spin accounting improves on the established spin counting technique by correcting for rapid T_1ρH relaxation and inefficient cross‐polarization. Spin accounting identifies three components: one that is well represented in CP spectra, one that is under‐represented in CP spectra due to rapid T_1ρH relaxation, and one that is under‐represented in CP spectra due to inefficient cross‐polarization. For a range of eight de‐ashed soils, C_obs was in the range 83–106%, indicating that virtually all potential signal could be accounted for after correcting for rapid T_1ρH relaxation and inefficient cross‐polarization. The second new technique, RESTORE (RE storation of S pectra via T _CH and T O ne R ho (T_1ρH) E diting), generates subspectra for the three components identified in spin accounting. The sum of the three RESTORE subspectra is essentially a corrected CP spectrum. The RESTORE spectra of all eight soils more closely resembled the corresponding, and presumably quantitative, Bloch decay spectra than did the CP spectra. RESTORE identifies the types of structures underestimated by CP, and the cause of their underestimation. Rapid T_1ρH relaxation most affected carbonyl and carbohydrate carbons, whereas inefficient cross‐polarization most affected aromatic carbons.     

Chemical structure of terrestrial humus materials formed from different vegetation characterized by solid-state 13C NMR with CP-MAS techniques

Eleven samples of terrestrial humus from different vegetational backgrounds were examined with solid-state ¹³C NMR using cross-polarization and magic-angle spinning (CP-MAS). In addition, all the samples were run with a dipolar dephasing pulse sequence for non-quarternary carbon suppression (NQS). The humus samples all appeared to contain small amounts of aromatic substances and larger amounts of aliphatic compounds. Most of the samples contained considerable amounts of hydroxyl groups and acetals, which originate mainly from carbohydrates. No correlations were found between vegetational background and chemical structure.     

Land-use related organic matter dynamics in North Cameroon soils assessed by 13C analysis of soil organic matter fractions

Topsoil samples from cultivated and adjacent non‐cultivated fields on three major agricultural soils in North Cameroon were fractionated into particle‐size fractions that were analysed subsequently for their C and ¹³C contents. The aim was to obtain further insight into the dynamics of soil organic matter (SOM) in relation to land use in Cameroon. Since organic carbon contents of the fractions were often very small, samples and analyses were extensively replicated to obtain robust statistical estimates of observed differences. For each soil type, differences in δ¹³C values between fields could be related to changes in the input and decomposition of organic matter arising from soil type, land management and, for example, the nature and abundance of weeds. Turnover of organic matter appeared to be fastest in the sand fraction, which is in line with results from earlier studies. In the finer fractions, clear differences in reaction to changes in input and decomposition were observed, that seem to be linked to differences in clay mineralogy. The results illustrate that SOM in the various fractions is much less stable and more strongly affected by changes in land use than might be assumed on the basis of changes in total SOM contents alone. At the same time, they demonstrate the relevance of ¹³C isotope analyses of SOM for studies on the impact of land use on these savannah soils with little SOM that are highly susceptible to degradation.     

Characteristic alterations of quantity and quality of soil organic matter caused by forest fires in continental Mediterranean ecosystems: a solid-state 13C NMR study

The variable effect of different types of forest fires on the quantity and quality of soil organic matter (SOM) was analysed by comparing burnt and unburnt soils from six forest ecosystems in central Spain by organic elemental analysis and solid‐state ¹³C nuclear magnetic resonance (NMR) spectroscopy. Whole soil samples were collected 1 to 2 years after the fires and included one site affected by two fires within 2 years. The fire‐affected soils showed no common pattern with respect to the amount of additional carbon (C_add) but at all sites, the fire enhanced the aromatic‐C content. The weakest fire intensity resulted in the greatest aromatic‐C enrichment factor, EF_{I(aromatic C)} indicating the greatest local accumulation of char. The respective C_add disclosed an EF_{I(aromatic C)} to EF_{I(alkyl C)} ratio, B_char, of c.1, which supports a small degree of charring. Extensive combustion and volatilization at stronger fire intensities yielded a decrease of EF_{I(aromatic C)} and an increase in B_char. These trends are in good agreement with fire intensity and forest fuel combustibility in the various sites and therefore these indices could be used to elucidate the quality and quantity of char input that occurs during and after forest fires. No ¹³C NMR evidence for substantial inputs from non‐charred plant necromass was found for any of the single‐burn soils. The large carboxyl‐C content of C_add is evidence of the occurrence of oxidation reactions very shortly after the fire. In comparing the single and double‐burn sites, no additional char input was observed for the double‐burn site, possibly because of complete combustion of young shrubs and char remains during the second fire. The large O‐alkyl‐C portion found in C_add of the double‐burn soil is best explained by decreased litter degradation.     

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.     

Fertilization effects on organic matter in physically fractionated soils as studied by 13C NMR: Results from two long-term field experiments

^l3C–nuclear magnetic resonance (NMR) spectra taken using magic–angle spinning (MAS), cross polarization (CP) and with total suppression of side bands (TOSS) are reported for soils from two long–term field experiments. One set of soils was from the Broadbalk Experiment at Rothamsted, UK (monoculture of winter wheat since 1843) and the other was from the Lermarken site of the Askov Long–Term Experiment on Animal Manure and Mineral Fertilizers (arable rotation since 1894). At both sites soil samples were taken from three fertilizer treatments: nil, inorganic fertilizers, animal manure. Spectra were obtained from whole soil samples and from the size fractions clay (<2 μrn), silt (2–20 μm) and, in some cases, sand (20–2000 μm). Comparison of the total strengths of the ¹³C–NMR signal for each size separate in relation to its total organic C content shows that clay, particularly, contains large percentages of C not detected by NMR because of the large magnetic susceptibilities of the soil minerals. It is proposed that the observed signals come from the more labile pools of soil organic matter (SOM), on the presumption that these pools are less closely associated with soil minerals and iron oxides and are likely to be less protected from microbial or enzymic decomposition. For both Rothamsted and Askov, functional groups in the 45–110 ppm region (N– and O–alkyls) dominate in the spectra for whole soils, with aromatics (110–160 ppm) and alkyls (0–45 ppm) signals being the next prominent. In the Askov whole soil samples ¹³C–NMR revealed no differences between nil, inorganic fertilizer and animal manure treatments but in the Rothamsted whole soil there were some small differences. Clay and silt fractions from Askov contain more alkyls and less aromatics than those from Rothamsted. For both sites clay in enriched in alkyls and depleted in aromatics relative to silt. Clay from Askov, but not Rothamsted, contains more N–alkyls (45–65 ppm) and less acetals (90–110 ppm) than silt. O–alkyls (65–90 ppm) account for more than 20% of the total signal in clay and silt from both sites. Fertilization regimes have not significantly affected the chemical composition of SOM associated with clay– and silt–sized fractions in the soils at either site. We conclude that the chemical composition of SOM is determined primarily by the interaction between the organisms responsible for decomposition and the mineral soil matrix rather than the nature of substrate input.     

Determination of refractory organic matter in marine sediments by chemical oxidation, analytical pyrolysis and solid-state 13C nuclear magnetic resonance spectroscopy

Seeking to quantify the amount of refractory organic matter (ROM), which includes black carbon‐like material (BC), in marine sediments, we have applied a two‐step procedure that consists of a chemical oxidation with sodium chlorite of the demineralized sediments followed by integration of the aromatic C region in the remaining residues by solid‐state ¹³C nuclear magnetic resonance (NMR) spectroscopy. The efficacy for lignin removal was tested by analytical pyrolysis in the presence of tetramethyl ammonium hydroxide (TMAH). Riverine, estuarine and offshore marine sediment samples were collected from the southwest Atlantic coast of Spain, a site of geological and environmental interest. Measured contents of BC‐like material ranged between 3.0 and 45.7% of the total organic carbon. Greater relative BC contents were found in riverine sediments close to urban areas, which show an elevated input of anthropogenic organic material. The contents of BC‐like material in offshore marine sediments (5.5–6.1%) were similar to those previously reported for these kinds of samples. However, NMR and pyrolysis‐GC/MS of the isolated ROM reveals that abundant refractory aliphatic organic material remains in most of the marine samples after chlorite oxidation. We suggest that this pool of aliphatic carbon may play an important role as a stable carbon pool within the global C cycle.     

Sludge-derived organic carbon in an agricultural soil estimated by 13C abundance measurements

The objective of this study is to develop a method to follow the dynamics of sludge‐derived organic carbon, which will allow us to understand the behaviour of trace metals in the sludge‐treated soils. We studied, in a sandy agricultural soil of southwest France, cultivated with maize and amended with sewage‐sludge over 20 years, the dynamics of different sources of organic matter and compared this with a control, which had never received any treatment. For the first time, a method is proposed that will distinguish and quantify sludge‐derived organic carbon, maize‐derived organic carbon, and native organic carbon. This method is based on the mean differences in δ¹³C abundances between native (−26.5‰), maize (−12.5‰) and sludge (−25.4‰) organic carbon. Three hypotheses on the dynamics of soil organic matter sources are proposed: (i) isotopic differences observed between control and sludge‐treated soils are due only to the incorporation of sludge C, whereas in the others, the control was used to model the incorporation of (ii) maize C or (iii) native C in the sludge‐treated soils. The comparison of the stocks of each source (native C, maize C and sludge C) found in the bulk soil with the sum of corresponding stocks found in particle‐size fractions allowed us to reject the two first hypotheses and to validate the last one. Repeated applications of sewage‐sludge induced accumulation of sludge‐derived organic carbon in the topsoil, and simultaneously contributed to the preservation of maize‐derived organic carbon. When sludge applications ceased, the rapid decrease in soil organic matter stocks was mostly caused by the degradation of the sludge‐derived organic carbon sources. At the same time, the maize‐derived organic carbon shifted from the coarsest fraction (200–2000 μm) to the finest fraction (0–50 μm). Therefore, this study has shown that repeated applications of sewage‐sludge induced changes in soil organic matter dynamics over time.     

Dynamics of soil organic matter turnover and soil respired CO2 in a temperate grassland labelled with 13C

The fate of carbon (C) in grassland soils is of particular interest since the vast majority in grassland ecosystems is stored below ground and respiratory C‐release from soils is a major component of the global C balance. The use of ¹³C‐depleted CO₂ in a 10‐year free‐air carbon dioxide enrichment (FACE) experiment, gave a unique opportunity to study the turnover of the C sequestered during this experiment. Soil organic matter (SOM), soil air and plant material were analysed for δ¹³C and C contents in the last year of the FACE experiment (2002) and in the two following growing seasons. After 10 years of exposure to CO₂ enrichment at 600 ppmv, no significant differences in SOM C content could be detected between fumigated and non‐fumigated plots. A ¹³C depletion of 3.4‰ was found in SOM (0–12 cm) of the fumigated soils in comparison with the control soils and a rapid decrease of this difference was observed after the end of fumigation. Within 2 years, 49% of the C in this SOM (0–12 cm) was exchanged with fresh C, with the limitation that this exchange cannot be further dissected into respiratory decay of old C and freshly sequestered new C. By analysing the mechanistic effects of a drought on the plant‐soil system it was shown that rhizosphere respiration is the dominant factor in soil respiration. Consideration of ecophysiological factors that drive plant activity is therefore important when soil respiration is to be investigated or modelled.     

Land-use effects on the composition of organic matter in particle-size separates of soils: II. CPMAS and solution 13C NMR analysis

Soils from A horizons of Eutrochrepts under spruce forest (Sf), mixed deciduous forest (Df), permanent grassland (Gp), and arable rotation (Ar) were fractionated into clay- (<2 μm), silt-(2–20 μm) and sand- (20–2000 μm) sized separates. ¹³C NMR spectroscopy was used to compare SOM composition across size separates and between land-use regimes. CPMAS ¹³C NMR spectroscopy showed that the intensity of signals assigned to carbohydrates (representing most O-alkyl C) and lignin (phenolic and methoxyl C) declined with decreasing particle size. Concurrently, alkyl C and C-substitution of aromatic C increased in the order sand, silt, clay. The amount of alkyl C correlated well with microbial resynthesis of carbohydrates. Solution ¹³C NMR spectra suggested that humic acids (HA) extracted from the size separates were richer in carboxyl C and aromatic C than the bulk size separates. Also HA reflected increasing percentage of alkyl C with decreasing particle size. O-alkyl C were lower in silt HA than in clay HA whereas aromatic C tended to peak in silt HA. These results suggested that sand-sized separates were enriched in plant residues (primary resources) whereas clay-sized separates were dominated by products of microbial resynthesis (secondary resources). Silt was rich in selectively preserved and microbially transformed primary resources. ¹³C NMR spectroscopy showed only small differences in SOM composition between land-use regimes, except that silt and silt HA from Ar were richer in aromatic C than those from the other plots. But enrichment factors (E= content in fraction/content in whole soil) revealed differences in the distribution of C species across the size separates. Relatively high E_aromatic (0.9) and E_o-alkyl (1.0) for sand from Gp indicated high amounts of plant residues, probably due to intense rhizodeposition and to occlusion of plant debris within aggregates. Low E_aromatic (0.3) and E_o-alkyl (0.3) for sand from Ar suggested depletion of primary resources, which could be attributed to disintegration of soil aggregates upon cultivation. A pronounced enrichment of alkyl C in Ar clay-sized separates (E_alkyl= 3.1) suggested large amounts of microbial carbon. Microbial products attached to clay surfaces by a variety of physico-chemical bondings appeared more stable against mineralization induced by cultivation than plant residues sequestered in aggregates.     

Following the decomposition of ryegrass labelled with 13C and 15N in soil by solid-state nuclear magnetic resonance spectroscopy

Investigating the biogeochemistry of plant material decomposition in soil has been restricted by difficulties extracting and identifying organic compounds. In this study the decomposition of ¹³C- and ¹⁵N-labelled Lolium perenne leaves mixed with mineral soil has been investigated over 224 days of incubation under laboratory conditions. Decomposition was followed using short-term rates of CO₂ evolution, the amounts of ¹³C and ¹⁵N remaining were determined by mass spectrometry, and ¹³C and ¹⁵N solid-state nuclear magnetic resonance (NMR) spectroscopy was used to characterize chemically the plant material as it decomposed. After 224 days 48% of the added ¹³C had been lost with a rapid period of C0₂ evolution over the first 56 days. The fraction of cross-polarization magic angle spinning (CP MAS) ¹³C NMR spectra represented by O-alkyl-C signal probably in carbohydrates (chemical shift, 60–90 p.p.m.) declined from 60 to 20% of the spectrum (chemical shift, 0–200 p.p.m.) over 224 days. The rate of decline of the total ¹³C exceeded that of the 60–90 p.p.m. signal during the first 56 days and was similar thereafter. The fraction of the CP MAS ¹³C NMR spectra represented by the alkyl- and methyl-C (chemical shift, 10–45 p.p.m.) signal increased from 5 to 14% over the first 14 days and was 19% after 224 days. CP MAS ¹³C NMR of ¹³C- and ¹⁵N-L. perenne contained in 100-μm aperture mesh bags incubated in the soil for 56 days indicated that the remaining material was mainly carbohydrate but there was an increase in the alkyl- and methyl-C associated with the bag's contents. After 224 days incubation of the labelled ¹³C- and ¹⁵N-L. perenne mixed with the soil, 40% of the added N had been lost. Throughout the incubation there was only one signal centred around 100 p.p.m. detectable in the CP MAS ¹⁵N NMR spectra. This signal corresponded to amide ¹⁵N in peptides and may have been of plant or microbial origin or both. Although there had been substantial interaction between the added ¹⁵N and the soil microorganisms, the associated redistribution of ¹⁵N from plant to microbial tissues occurred within the amide region. The feasibility of following some of the component processes of plant material decomposition in soil using NMR has been demonstrated in this study and evidence that microbial synthesis contributes to the increase in alkyl- and methyl-C content of soil during decomposition has been represented.     

13C solid-state NMR assessment of decomposition pattern during co-composting of sewage sludge and green wastes

The fate of organic matter during composting is poorly understood. Therefore, we analysed composts of sewage sludges and green wastes (44 samples representative of 11 stages of biodegradation) by conventional chemical methods: pH, humic (HA) and fulvic acid (FA) content, C, N and organic matter (OM) content, and by ¹³C CPMAS NMR to assess the decomposition process of the organic matter. Chemical changes clearly occurred in two phases: first, decomposition of OM during the first 2 months was characterized by decreased C/N ratios, OM content and increased pH; and second, a humification process with increased HA/FA ratios. NMR spectrum changes confirmed this pattern, with an increase in aromaticity and a decrease in alkyl C. A decrease of syringyl to guaiacyl ratio (S/G), a sign of lignin transformation, also indicated humification during composting. NMR spectroscopic properties of composts were also studied by means of principal components analysis (PCA) and revealed changes according to the degree of compost maturation. The factorial map presents a chronological distribution of composts on the two first principal components. The influences of eight chemical factors on the PCA ordination of composts as monitored by their evolution by NMR were also studied by multivariate analyses. PCA clearly indicated two phases: the rapid decomposition of organic matter followed by the formation of humic‐like substances. The first phase, that is ‘new’ composts, was strongly correlated with OM contents, pH and C/N ratios whereas the second phase, corresponding to ‘old’ compost, was correlated with pH, HA content and HA/FA ratio. These results confirm that knowledge of the formation of humic substances is indispensable to suitable monitoring of the composting process.     

Dissolved organic matter sorption on sub soils and minerals studied by 13C-NMR and DRIFT spectroscopy

Sorption on the mineral matrix is an important process restricting the movement of dissolved organic matter (DOM) in soils. In this study, we aimed to identify the chemical structures responsible for the retention of DOM by sorption experiments with total DOM and acidic humic substances (AHS), containing humic and fulvic acids, on soil samples and minerals (goethite, ferrihydrite, and amorphous Al(OH)₃). The AHS remaining in solution after sorption were studied by ¹³C nuclear magnetic resonance (NMR) analysis, and total DOM and AHS for bed on the surfaces of minerals by diffuse reflectance Fourier-transform infrared (DRIFT) spectroscopy. The soil samples were taken from strongly sorbing Bw horizons of two Inceptisols rich in pedogenetic Fe (29 and 35 g kg ^?1) and containing little C (7 and 22 g kg^?1). The ¹³C-NMR spectra showed that sorption causes a preferential removal of aromatic and carboxyl C from the solution, whereas alkyl-C accumulates in the solution. No change was observed for O-alkyl C. The DRIFT spectra of sorbed total DOM and AHS showed a relative increase of the band intensity of carboxyl groups compared to DOM in the initial solution, confirming the importance of those groups for the sorption to mineral surfaces. The spectra also indicated reactions of carboxyl groups with metals at the mineral surfaces. The extent to which the carboxyl groups are bound depended on the surface coverage with DOM and the type of mineral.     

13C NMR studies of organic matter in whole soils: H. A case study of some Rothamsted soils

Nuclear magnetic resonance (NMR) spectra were obtained for solid samples of whole soils from three long–term field sites at Rothamsted Experimental Station, UK. In all sites, soil organic matter content was either increasing or decreasing due to contrasted long–continued treatments. Two soils were from Highfield, one from under old grassland (47 g organic C kg^?1) and one from an area kept as bare fallow following ploughing of grass 21 years previously (14 g organic C kg^?1). Three soils were taken from Broadbalk, two from plots within the Broadbalk Continuous Wheat Experiment which had received no fertilizer or animal manure annually for 148 years (7 and 27 g organic C kg^?1, respectively) and one from Broadbalk Wilderness, wooded section (38 g organic C kg^?1). Broadbalk Wilderness was arable until 1881 and has reverted to deciduous woodland in the subsequent 110 years. Two soils were from Geescroft, one from an arable field (9 g organic C kg^?1) and one from Geescroft Wilderness (35 g organic C kg^?1) which began reversion to deciduous woodland at the same time as Broadbalk Wilderness but is now acid (pH = 4.2) in contrast to Broadbalk which is calcareous (pH = 7.3). Solid–state ¹³C NMR spectra were obtained on a 300–MHz instrument using cross polarization (CP) and magic angle spinning (MAS). All samples exhibited peaks in the following spectral regions: 0–45 ppm (alkyl), 45–60 ppm (methoxyl, carbohydrate and derivatives), 60–110 ppm (carbohydrates and derivatives, C–α of peptides), 110–160 ppm (aromatics) and 160–185 ppm (carboxyl groups and derivatives). Within the spectrum of a specific sample it was not possible to determine the relative proportions of soil organic carbon in the different forms identified because a range of factors can potentially alter the relative areas of peaks in different regions of the spectrum. However, from a comparison of relative peak areas within a set of soils from a given site, differing only in organic matter content, information can be deduced regarding the forms of C that are more or less subject to change in response to land use or management. At all sites carbohydrate C appears to be the form that is most subject to change, suggesting that it is an ‘active’ fraction compared with the other forms. It was greatest where organic matter inputs were greatest (due to inputs of farmyard manure or reversion to woodland) and declined relative to other forms following ploughing of old grassland. Alkyl C increased as total C accumulated but did not decline relative to other forms following ploughing of grass. One reason for the non–quantitative nature of the soil ¹³C CPMAS spectra was a short (approximately 1 ms) component of the rotating–frame T_I relaxation time for H nuclei (T_1pH). This problem was not overcome by acquiring data at – 60°C. In principle, solid–state spectra of soils obtained by direct polarization (i.e. without CP) might produce quantitative results, but the low C content of most mineral soils (10–50 g C kg^?1) precludes this, given current instrumentation.     

13C NMR of humic substances: pH and solvent effects

Since the concentration of free radicals in humic subtances increases at high pH the use of basic solutions for ¹³C NMR spectroscopy may cause broadening and loss of aromatic signals, with distortion of intensity distributions. No such effects were found in ¹³C spectra of soil humic and fulvic acid, an aquatic fulvic acid, and two phenolic polymers run in aqueous solutions at different pH values, and in dimethylsulphoxide. With increasing pH, the peak in the carboxyl region shifted in a manner consistent with greater dissociation of carboxyl and phenolic groups, and also certain features in the aliphatic and carboxyl regions were enhanced under some solution conditions. Elevated solution temperature (70°C) caused only slight improvement in the resolution of some lines. Chemical shifts were determined for some known phenolic and benzenecarboxylic acid compounds in DMSO and NaOD. The range for phenolic carbons extended to 173 ppm in NaOD, while some aromatic carbons occurred around 105 ppm, in the same region as anomeric carbons. Thus, even under quantitative acquisition conditions, relative areas may be used only to estimate proportions of different types of carbons and functional groups.