Characterization Of Soil Organic Matter Research Articles

Improved Characterization of Soil Organic Matter by Integrating FT-ICR MS, Liquid Chromatography Tandem Mass Spectrometry, and Molecular Networking: A Case Study of Root Litter Decay under Drought Conditions.

Understanding of how soil organic matter (SOM) chemistry is altered in a changing climate has advanced considerably; however, most SOM components remain unidentified, impeding the ability to characterize a major fraction of organic matter and predict what types of molecules, and from which sources, will persist in soil. We present a novel approach to better characterize SOM extracts by integrating information from three types of analyses, and we deploy this method to characterize decaying root-detritus soil microcosms subjected to either drought or normal conditions. To observe broad differences in composition, we employed direct infusion Fourier-transform ion cyclotron resonance mass spectrometry (DI-FT-ICR MS). We complemented this with liquid chromatography tandem mass spectrometry (LC-MS/MS) to identify components by library matching. Since libraries contain only a small fraction of SOM components, we also used fragment spectral cosine similarity scores to relate unknowns and library matches through molecular networks. This integrated approach allowed us to corroborate DI-FT-ICR MS molecular formulas using library matches, which included fungal metabolites and related polyphenolic compounds. We also inferred structures of unknowns from molecular networks and improved LC-MS/MS annotation rates from ∼5 to 35% by considering DI-FT-ICR MS molecular formula assignments. Under drought conditions, we found greater relative amounts of lignin-like vs condensed aromatic polyphenol formulas and lower average nominal oxidation state of carbon, suggesting reduced decomposition of SOM and/or microbes under stress. Our integrated approach provides a framework for enhanced annotation of SOM components that is more comprehensive than performing individual data analyses in parallel.

Unravelling the thermodynamic properties of soil ecosystems in mature beech forests

Thermodynamics is a vast area of knowledge with a debatable role in explaining the evolution of ecosystems. In the case of soil ecosystems, this role is still unclear due to difficulties in determining the thermodynamic functions that are involved in the survival and evolution of soils as living systems. The existing knowledge is largely based on theoretical approaches and has never been applied to soils using thermodynamic functions that have been experimentally determined. In this study, we present a method for the complete experimental thermodynamic characterization of soil organic matter. This method quantifies all the thermodynamic functions for combustion and formation reactions which are involved in the thermodynamic principles governing the evolution of the universe. We applied them to track the progress of soil organic matter with soil depth in mature beech forests. Our results show that soil organic matter evolves to a higher degree of reduction as it is mineralized, yielding products with lower carbon but higher energy content than the original organic matter used as reference. These products have higher entropy than the original one, demonstrating how the soil ecosystem evolves with depth, in accordance with the second law of thermodynamics. The results were sensitive to soil organic matter transformation in forests under different management, indicating potential applicability in elucidating the energy strategies for evolution and survival of soil systems as well as in settling their evolutionary states.

Disentangling the effects of temperature and reactive minerals on soil carbon stocks across a thermal gradient in a temperate native forest ecosystem

Effects of global warming on soil organic carbon (C) can be investigated by comparing sites experiencing different temperatures. However, observations can be affected by covariance of temperature with other environmental properties. Here, we studied a thermal gradient in forest soils derived from volcanic materials on Mount Taranaki (New Zealand) to disentangle the effects of temperature and reactive minerals on soil organic C quantity and composition. We collected soils at four depths and four elevations with mean annual temperatures ranging from 7.3 to 10.5 °C. Soil C stocks were not significantly different across sites (average 162 MgC ha−1 to 85 cm depth, P > .05). Neither aluminium (Al)-complexed C, nor mineral-associated C changed significantly (P > .05) with temperature. The molecular characterisation of soil organic matter showed that plant-derived C declined with increasing temperature, while microbial-processed C increased. Accompanying these changes, soil short-range order (SRO) constituents (including allophane) generally increased with temperature. Results from structural equation modelling revealed that, although a warmer temperature tended to accelerate soil organic C decomposition as inferred from molecular fingerprints, it also exerted a positive effect on soil total C presumably by enhancing plant C input. Despite a close linkage between mineral-associated C and soil organic C, the increased abundance of reactive minerals at 30–85 cm depth with temperature did not increase soil organic C concentration at that depth. We therefore propose that fresh C inputs, rather than reactive minerals, mediate soil C responses to temperature across the thermal gradient of volcanic soils under humid-temperate climatic conditions.

Chemical characterization of soil organic matter in differents management practices in the Cerrado-Pantanal ecotone

Different land uses and occupations have a direct impact on edaphic attributes, which can be identified by soil quality indicators, with organic matter being an excellent edaphic quality indicator. The objective of the work was to chemically characterize soil organic matter (SOM) in different management systems in the municipality of Aquidauana, Mato Grosso do Sul, Brazil. Soil samples were collected in five areas with different management practices, in addition to an area of native Cerrado forest (NF), and the managed areas were: exposed soil (ES), conventional tillage system (CTS), no-tillage (NTS), permanent pasture (PP) and sugarcane (SC). In addition to the determination of bulk density (Bd), total organic carbon (TOC) contents and stocks were quantified, as well as carbon (C) contents and stocks of the humified fractions: humic acid (AH), fulvic acid (AF) and humin (HUM), with subsequent calculation of HA/FA and alkaline extract/humin (AE/HUM) and TOC stratification index (SI). There were no significant changes for Ds after six years of implementation of management systems. TOC levels and stocks were higher in PP and NTS areas. The HUM fraction was predominant, with higher contents and stocks of all fractions for the PP and NTS areas, demonstrating the greater intensity of the humification process in these management systems, with greater stability of C in the soil after six years of experiment. The PCA indicated a difference between the areas that present conservation practices in the ES and CTS areas. The NF, NTS, PP and SC areas contribute to SOM stabilization.

Electrochemical Framework for Dynamic Tracking of Soil Organic Matter (SOM)

Soil Health parameters serve as excellent surrogate measures towards assessing environmental quality and understanding effects of climate change mitigation via carbon sequestration. Soil health monitoring in the near past has been highly qualitative and speculative with more recent advancements still trying to fill the void of determining a holistic soil profile through phantom based measurements. In fields and crop-circles, testing methods to perform in-situ profiling however from one lab is likely to vary with another and hence results are not universally compatible. Additionally, these normalized techniques ideally involve empirical approaches, extensive sample preparation which adds on to a temporal factor along with equipment for extraction and subsequently-analysis. Even with the upcoming research breakthroughs in non-destructive approaches like spectroscopy and tomography-based models, there is a barrier in-terms of equipment complexity and availability as well as high costs and high logistical overhead.There are a wide number of redox systems present in soil that exist in reduced state under ideal conditions (submerged). While Carbon monitoring would be highly beneficial in the field, an overall sense of the organic matter nutrients present in soil denoted as soil organic matter understanding would offer a wholistic manner of surveying soil and environmental health. In this work, faradaic and non-faradaic measurements are utilized to determine threshold changes in soil electrochemical activity due to presence of various electroactive substances present in the matrix. First, a galvanostatic approach using the chronopotentiometry modality was applied to a soil slurry system- ‘test’ sample to which different levels (w/v %) of the following amendments were added to see how the effective charge modulation can be visualized. Utilizing this experimental framework as preliminary information- A hypothesized mechanism of interaction between the RTIL (Room Temperature Ionic-Liquid) modified electrode and the OM functional moieties based on hydrogen bonding and pi-pi interactions captured using charge-based (chronocoulometry) and interfacial mapping-based (electrochemical impedance spectroscopy) method is utilized to design and test a first-of-a-kind electrochemical soil organic matter (SOM) sensor. Figure 1. Chronopotentiometric characterization of soil organic matter using an RTIL modified electrode. Figure 1

Reproducibility of Rock-Eval® thermal analysis for soil organic matter characterization

Rock-Eval® (RE) is a thermal analysis technique increasingly used to characterise soil organic matter. To interpret the results, particularly when investigating differences between samples, it is necessary to know the expected ranges of analytical error associated with the RE measurements. Moreover, the RE analyzer is now at its seventh version (RE7) while most literature results were produced using the previous version (RE6). Thus, a characterization of the reproducibility of RE measurements is necessary. We measured the reproducibility of RE measurements using fifteen samples from French croplands and forests that were analysed on five different RE instruments, located in different laboratories and belonging to both generations RE6 and RE7. From each RE analysis, we extracted RE parameters commonly used for soil organic matter characterization and we performed the prediction of the active and stable soil organic carbon fractions using a machine learning model (PartySOC) that uses RE parameters. We obtained a measure of the expected relative errors in RE parameters and PartySOC predictions per instrument, across instruments of the same generation and across generations. We found that the parameters total organic carbon (TOC), mineral carbon (MinC) and R-index are well reproducible, even across the RE6 and RE7 generations. Instead, the hydrogen index (HI) and oxygen index (OI) are more sensitive to signal variations, even within the same generation, especially when TOC is low. The PartySOC predictions were well reproducible across RE6 instruments but not across RE generations. In the future, the results of this study will help discriminate relevant differences between soil samples characterised using RE thermal analysis.

Soil pH and Soluble Organic Matter Shifts Exerted by Heating Affect Microbial Response

Fire-induced alterations to soil pH and organic matter play an important role in the post-fire microbial response. However, the magnitude of which each parameter affects this response is still unclear. The main objective of this work was to determine the magnitude in which soil pH and organic matter fire-induced alterations condition the response of viable and cultivable micro-organisms using laboratory heating, mimicking a range of fire intensities. Four heating treatments were applied to unaltered forest soil: unheated, 300, 450, and 500 °C. In order to isolate the effect of nutrient or pH heating-induced changes, different culture media were prepared using soil:water extracts from the different heated soils, nutrient, and pH amendments. Each medium was inoculated with different dilutions of a microbial suspension from the same original, unaltered soil, and microbial abundance was estimated. Concurrently, freeze-dry aliquots from each soil:water extract were analyzed by pyrolysis-gas chromatography/mass spectrometry. The microbial abundance in media prepared with heated soil was lower than that in media prepared with unheated soil. Nutrient addition and pH compensation appear to promote microbial proliferation in unaltered and low-intensity heated treatments, but not in those heated at the highest temperatures. Soil organic matter characterization showed a reduction in the number of organic compounds in soil-heated treatments and a marked increase in aromatic compounds, which could be related to the observed low microbial proliferation.

Advances in Molecular and Microscale Characterization of Soil Organic Matter: Current Limitations and Future Prospects.

Soil organic matter (SOM) comprises a continuum of organic materials from granular organic debris to small organic molecules and contains more organic carbon than global vegetation and the atmosphere combined. It has remarkable effects on soil ecological functions and the global carbon cycle as well as the fate of pollutants in the terrestrial ecosystem. Therefore, characterization of SOM is an important topic in soil science, ecology, and environmental science. Chemical complexity and spatial heterogeneity are by far the two biggest challenges to our understanding of SOM. Recent developments in analytical techniques and methods provide the opportunity to reveal SOM composition at the molecular level and to observe its distribution in soils at micro- and nanoscales, which have greatly improved our understanding of SOM. This paper reviews the outstanding advances in SOM characterization regarding these two issues from target and nontarget analyses comprising molecular marker analysis, ultrahigh-resolution mass spectrometry analysis, and in situ microscopic imaging techniques such as synchrotron-based spectromicroscopy, nanoscale secondary ion mass spectrometry, and emerging electron and optical microscopic imaging techniques. However, current techniques and methods remain far from unlocking the unknown properties of SOM. We systematically point out the limitations of the current technologies and outline the future prospects for comprehensive characterization of SOM at the molecular level and micro- and nanoscales, paying particular attention to issues of environmental concern.

The influence of organic and inorganic nutrient inputs on soil organic carbon functional groups content and maize yields

Locally available organic inputs to soil, solely or in combination with inorganic fertilizers, are used to reverse declining soil fertility and improve soil organic matter content (SOM) in smallholder farms of most Sub-Saharan Africa (SSA) countries. Soil organic matter characterization can indicate soil organic input, carbon (C) sequestration potential, or even an authentication tool for soil C dynamics in C stocks accounting. This study determined the effects of the long-term application of selected integrated soil fertility management (ISFM) technologies on SOM functional group composition and maize yields. The study was carried out on an ongoing long-term soil fertility field experiment established in 2004 in Mbeere South sub-county, the drier part of upper Eastern Kenya. The experimental design was a randomized complete block design. The ISFM treatments were 60 kg ha−1 nitrogen (N) from goat manure (GM60); 30 kg ha−1 inorganic N fertilizer (IF30); 60 kg ha−1 inorganic N fertilizer (IF60); GM30+IF30; 90 kg ha−1 inorganic N fertilizer (IF90); 60 kg ha−1 N from lantana (Lantana camara) (LC60); LC30+IF30; 60 kg ha−1 N from mucuna beans (Mucuna pruriens) (MP60); MP30+IF30; 60 kg ha−1 N from Mexican sunflower (Tithonia diversifolia) (TD60); TD30+IF30, and a control with no inputs. The C compositions of ground soil samples and organic amendments were analyzed using 13C solid-state NMR. The GM60, GM30+IF30, LC60, and TD60 treatments had much higher Alkyl and O-Alkyl C SOM functional groups than the control and other treatments. The average soil C for the control was 7.47 mg kg−1 and ranged from 5.03 to 7.37, 9.57 to 18.77, and 7.03–14.50 mg kg−1 for inorganic fertilizers, organic fertilizers, and organic + inorganic fertilizers, respectively. The mean grain yield for the control was 0.56 Mg ha−1 and ranged from 1.51 to 1.99, 1.94 to 4.16, and 2.98–4.60 Mg ha−1 for inorganic fertilizers, organic fertilizers, and organic + inorganic fertilizers, respectively. The results showed that a long-term application of sole organic fertilizers or combined with inorganic fertilizers increases maize yield and soil C sequestration potential. The increase was attributed to high Alkyl and O-Alkyl C SOM functional groups. Hence, knowing the C fraction content of organic inputs is vital in determining the best-fit management technologies for ameliorating soil fertility and sustaining and/or improving crop yields.

Soil Organic Matter Characterization by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FTICR MS): A Critical Review of Sample Preparation, Analysis, and Data Interpretation.

The biogeochemical cycling of soil organic matter (SOM) plays a central role in regulating soil health, water quality, carbon storage, and greenhouse gas emissions. Thus, many studies have been conducted to reveal how anthropogenic and climate variables affect carbon sequestration and nutrient cycling. Among the analytical techniques used to better understand the speciation and transformation of SOM, Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) is the only technique that has sufficient mass resolving power to separate and accurately assign elemental compositions to individual SOM molecules. The global increase in the application of FTICR MS to address SOM complexity has highlighted the many challenges and opportunities associated with SOM sample preparation, FTICR MS analysis, and mass spectral interpretation. Here, we provide a critical review of recent strategies for SOM characterization by FTICR MS with emphasis on SOM sample collection, preparation, analysis, and data interpretation. Data processing and visualization methods are presented with suggested workflows that detail the considerations needed for the application of molecular information derived from FTICR MS. Finally, we highlight current research gaps, biases, and future directions needed to improve our understanding of organic matter chemistry and cycling within terrestrial ecosystems.

Molecular characterization of soil organic matter and its extractable humic fraction from long-term field experiments under different cropping systems

Soil Organic Matter (SOM) is essential for soil stability, fertility and crop productivity. Recent findings showed that SOM supramolecular structure is strongly influenced by cultivation and land management. In this study, we investigated the molecular dynamics of organic matter in soils from three long-term field-experiments which were subjected for 20 consecutive years to the following crop managements: i) non cultivated (Control); ii) maize mono-culture; and iii) maize-leguminous (Vicia Faba) rotation. Off-line pyrolysis TMAH-GC-MS (thermochemolysis) and solid-state 13C NMR spectroscopy were applied for the direct molecular characterization of OM components in both the bulk soils and their humic extracts. While 20 consecutive years of Maize mono-cultivation led to alteration of the SOM hydrophobic composition, with a decrease in alkyl and aliphatic compounds (e.g. 47.4% reduction in fatty acids), and an increase in hydrophilic labile components (42.2%), the crop-rotated soils showed a partial preservation of the pristine SOM composition by maintaining the content of hydrophobic and lipid constituents (only 1.8% reduction). Our results suggest that different cropping systems change the SOM molecular dynamics and stability in long-term field experiments by primarily altering the hydrophobic components of SOM. In particular, Maize mono-cultivation leads to a progressive degradation of SOM quality. Yet, the introduction of a leguminous species in crop rotation with Maize maybe an advantageous strategy to preserve SOM quality while reducing SOC losses.

Chemical composition of soil organic matter and potential enzyme activity in the topsoil along a moisture gradient in the High Arctic (Svalbard)

The soil active layer in the High Arctic is a reservoir for nutrients and microbial communities, but their behavior and interactions remain unclear. Therefore, the objective of this study was to characterize soil organic matter (SOM) composition and potential enzyme activities in soil active layer of Svalbard. Samples from two different sites and two different soil layers (biocrust: 0–1 cm; mineral soil: 5–10 cm depth) were collected along a moisture gradient. The molecular SOM composition was assessed using pyrolysis-field ionization mass spectrometry (Py-FIMS), and the potential activities of seven extracellular enzymes characteristic for different soil microbial processes were measured. In general, SOM in both sites was dominated by lipids/sterols, alkylaromatics and phenols/lignin monomers. Dehydroergosterol prevailed in a majority of the samples. Biocrusts samples in both sites had higher total ion intensity and volatile matter, but lower lipid and alkylaromatics contents. The potential enzyme activities varied significantly across the sites, depths and moisture gradients and, in general, were higher in the biocrusts than in underlying soils. Besides, majority of measured enzyme activities were lower in the wetter plots. Taken together, our results expand the knowledge of SOM and enzyme activity in High Arctic topsoil, which is important for better understanding the dynamic of soil biotic and abiotic processes in polar environments, particularly in the context of climate change.

Do Fulvic, Humic, and Humin Carbon Fractions Represent Meaningful Biological, Physical, and Chemical Carbon Pools?

Humic fractions have been used to characterize soil organic matter chemistry and to identify different carbon (C) pools. The relevance of alkaline fractionation is questioned because of probable artifacts created by the extraction. Fractionation using water‐soluble organic C (WSOC), soil microbial biomass C (SMBC), permanganate‐oxidizable C (POX‐C), and particulate organic matter C (POM‐C) methodologies are sensitive to C changes. Mineral‐associated organic C (MAOC) includes WSOC and other C fractions that are not all passive C. We measured 15 proficiency soils ranging in texture, mineralogy, pH, and soil organic C (SOC) to isolate C pools with differing turnover times. We define WSOC, SMBC, POX‐C, and fulvic acid (FA) as fast pools. Slow cycling pools include POM‐C and humic acid (HA) fractions, whereas passive pools include MAOC minus WSOC (PMAOC) and humin C (HC). Data showed FA was correlated with POX‐C (r = 0.84) as a fast C pool but was slightly more correlated with PMAOC (r = 0.86), a passive pool. Slow C pools of HA and POM‐C were not strongly correlated (r = 0.49). All alkaline fractions of C showed highest correlations to PMAOC, followed by POX‐C, and all showed poor correlations with POM‐C. The conventional first extraction fraction (A) yielded 70% recovery of the total FA extracted, whereas only 45% of HA was obtain. Acid soils on average had higher total FA. The regression model using physical fractionation was the best (SOC = WSOC + POM‐C + PMAOC, R2 0.98). Only the HA fraction was a significant parameter in the regression of SOC = FA + HA + HC (p = 0.0029).Core Ideas Humic acid C showed the strongest correlation to all C pools except POM‐C. Recovery of fulvic C in conventional fraction was 70% of a total of three fractions. Recovery of humic acid C in conventional fraction was 45% of a total of three fractions. The distribution of fulvic (FA) to humic acid (HA) C changed with acid soils having higher FA. Regression of soil organic C = FA + HA + humin only showed the HA to be significant.

Molecular features of organic matter in anthropogenic earthen mounds, canals and lagoons in the Pago Lindo archaeological complex (Tacuarembó, Uruguayan lowlands) are controlled by pedogenetic processes and fire practices

The Pago Lindo site in the River Plate basin (Uruguay) is an important pre-hispanic mound settlement (ca. 3000–600 BP), of which the technologically and socially advanced nature have only recently been revealed. Different angles of pedogenetic and palaeo-ecological science are rapidly improving our understanding of the history of the site and the relationships between human activity and habitat alteration. Here we add to this progress by molecular characterization of soil organic matter (SOM) in anthropogenic earthen mounds, canals and lagoons from Pago Lindo, by pyrolysis-GC–MS. The results showed that the SOM in the earthen mound and the archaeological lagoon are composed of microbial (partially chitin), aliphatic (partially root-derived) and pyrogenic (from fireplaces or wildfires) materials, whereas the canal and younger lagoon contained SOM with larger proportions of relatively intact plant remains (lignin and polysaccharides from herbaceous species). In most systems, a clear degradation trend can be observed with increasing depth –either from intact plant remains to microbial tissues (canal, young lagoon), or from microbial tissues to recalcitrant aliphatic and pyrogenic sources (earthen mound, ancient lagoon)– which is indicative of a strong control of decay intensity on SOM composition. Multivariate statistics confirmed that most variability in pyrolysis fingerprints can be attributed to degradation/preservation dynamics, which probably erased most of the molecular information on habitat development under the influence of past societies, including nearby maize cultivation.

Stable carbon and nitrogen isotopic composition of leaves, litter, and soils of various ecosystems along an elevational and land-use gradient at Mount Kilimanjaro, Tanzania

Abstract. Variations in the stable isotopic composition of carbon (δ13C) and nitrogen (δ15N) of fresh leaves, litter, and topsoils were used to characterize soil organic matter dynamics of 12 tropical ecosystems in the Mount Kilimanjaro region, Tanzania. We studied a total of 60 sites distributed along five individual elevational transects (860–4550 m a.s.l.), which define a strong climatic and land-use gradient encompassing semi-natural and managed ecosystems. The combined effects of contrasting environmental conditions, vegetation, soil, and management practices had a strong impact on the δ13C and δ15N values observed in the different ecosystems. The relative abundance of C3 and C4 plants greatly determined the δ13C of a given ecosystem. In contrast, δ15N values were largely controlled by land-use intensification and climatic conditions. The large δ13C enrichment factors (δ13Clitter − δ13Csoil) and low soil C∕N ratios observed in managed and disturbed systems agree well with the notion of altered SOM dynamics. Besides the systematic removal of the plant biomass characteristic of agricultural systems, annual litterfall patterns may also explain the comparatively lower contents of C and N observed in the topsoils of these intensively managed sites. Both δ15N values and calculated δ15N-based enrichment factors (δ15Nlitter − δ15Nsoil) suggest the tightest nitrogen cycling at high-elevation (> 3000 m a.s.l.) ecosystems and more open nitrogen cycling both in grass-dominated and intensively managed cropping systems. However, claims about the nature of the N cycle (i.e. open or closed) should not be made solely on the basis of soil δ15N as other processes that barely discriminate against 15N (i.e. soil nitrate leaching) have been shown to be quite significant in Mount Kilimanjaro's forest ecosystems. The negative correlation of δ15N values with soil nitrogen content and the positive correlation with mean annual temperature suggest reduced mineralization rates and thus limited nitrogen availability, at least in high-elevation ecosystems. By contrast, intensively managed systems are characterized by lower soil nitrogen contents and warmer conditions, leading together with nitrogen fertilizer inputs to lower nitrogen retention and thus significantly higher soil δ15N values. A simple function driven by soil nitrogen content and mean annual temperature explained 68 % of the variability in soil δ15N values across all sites. Based on our results, we suggest that in addition to land-use intensification, increasing temperatures in a changing climate may promote soil carbon and nitrogen losses, thus altering the otherwise stable soil organic matter dynamics of Mount Kilimanjaro's forest ecosystems.

Improving Infrared Spectroscopy Characterization of Soil Organic Matter with Spectral Subtractions.

Soil organic matter (SOM) underlies numerous soil processes and functions. Fourier transform infrared (FTIR) spectroscopy detects infrared-active organic bonds that constitute the organic component of soils. However, the relatively low organic matter content of soils (commonly < 5% by mass) and absorbance overlap of mineral and organic functional groups in the mid-infrared (MIR) region (4,000-400 cm-1) engenders substantial interference by dominant mineral absorbances, challenging or even preventing interpretation of spectra for SOM characterization. Spectral subtractions, a post-hoc mathematical treatment of spectra, can reduce mineral interference and enhance resolution of spectral regions corresponding to organic functional groups by mathematically removing mineral absorbances. This requires a mineral-enriched reference spectrum, which can be empirically obtained for a given soil sample by removing SOM. The mineral-enriched reference spectrum is subtracted from the original (untreated) spectrum of the soil sample to produce a spectrum representing SOM absorbances. Common SOM removal methods include high-temperature combustion ('ashing') and chemical oxidation. Selection of the SOM removal method carries two considerations: (1) the amount of SOM removed, and (2) absorbance artifacts in the mineral reference spectrum and thus the resulting subtraction spectrum. These potential issues can, and should, be identified and quantified in order to avoid fallacious or biased interpretations of spectra for organic functional group composition of SOM. Following SOM removal, the resulting mineral-enriched sample is used to collect a mineral reference spectrum. Several strategies exist to perform subtractions depending on the experimental goals and sample characteristics, most notably the determination of the subtraction factor. The resulting subtraction spectrum requires careful interpretation based on the aforementioned methodology. For many soil and other environmental samples containing substantial mineral components, subtractions have strong potential to improve FTIR spectroscopic characterization of organic matter composition.

Online supercritical fluid extraction mass spectrometry (SFE-LC-FTMS) for sensitive characterization of soil organic matter.

We report a novel technical approach for subcritical fluid extraction (SFE) for organic matter characterization in complex matrices such as soil. The custom platform combines on-line SFE with micro-solid phase extraction, nano liquid chromatography (LC), electrospray ionization and Fourier transform mass spectrometry (SFE-LC-FTMS). We demonstrated the utility of SFE-LC-FTMS, including results from both Orbitrap and FTICR MS, for analysis of complex mixtures of organic compounds in a solid matrix by characterizing soil organic matter in peat, a high-carbon soil. For example, in a single experiment, >6000 molecular formulas can be assigned based upon FTICR MS data from 1-50 μL of soil samples (roughly 1-50 mg of soil, dependent on soil density), nearly twice that typically obtained from direct infusion liquid solvent extraction (LSE) from an order of magnitude larger volume of the same soil. The detected species consisted predominately of lipid-like, lignin-like and protein-like compounds, based on their O/C and H/C ratios, with predominantly CHO and CHONP molecular compositions. These results clearly demonstrate that SFE has the potential to effectively extract a variety of molecular species and could become an important member of a suite of extraction methods for studying SOM and other natural organic matter. This is especially true when comprehensive coverage, minimal sample volumes, and high sensitivity are required, or when the presence of organic solvent residue in residual soil is problematic. The SFE based extraction protocol could potentially enable spatially resolved characterization of organic matter in soil with a resolution of ∼1 mm3 to facilitate studies probing the spatial heterogeneity of soil.

Coupling pyrolysis with mid-infrared spectroscopy (Py-MIRS) to fingerprint soil organic matter bulk chemistry

A novel method coupling pyrolysis with mid-infrared spectroscopy (Py-MIRS) was developed to characterize soil organic matter (SOM) chemistry in soils. The pyrolyzer was interfaced to the MIR spectrometer by means of a Brill cell™ (CDS Analytica). The set-up generates pyrolysis fingerprint spectra from which individual pyrolysis products can be related to SOM bulk chemistry. Py-MIRS development involved the testing of experimental conditions like pyrolysis temperature (550, 700, 1000 °C), heating rate (20 °C s−1 and 20 °C ms−1) and time (15, 30 and 60 s) using reference standard compounds ranging from carbohydrates to phenols varying in chemical and structural composition like levoglusogan, gluten, tannin, syringol, pectin and leucine falling within different compound categories (carbohydrates, amino acids, proteins, phenols, etc.) as well as soil samples. Pyrolysis yields of prominent specific functional groups, like aliphatics (CH stretching at 2930 cm−1) and CC aromatics (1510 cm−1), varied with pyrolysis temperature, heating rate and time. The preferred settings for high pyrolysis yield and minimized secondary reactions were obtained at a pyrolysis temperature of 700 °C, heating rate of 20 °C ms−1 and heating time of 30 s. The suitability of Py-MIRS to detect changes in SOM composition was evaluated by comparing Py-MIRS results to Diffuse Reflectance Fourier Transform mid-Infrared Spectroscopy (DRIFTS) results. Soil samples taken from the Static Fertilization Experiment, Bad Lauchstädt, Germany (Chernozem) revealed a major SOM contribution of the peak at 1750 cm−1 (CO), followed by peaks at 950 (CH), 1510 (CC), 1176 (CH, OH) cm−1, with smaller contributions from the 2930 (CH) and 3015 (CH4) cm−1 peaks, apart from a dominant CO2 peak. Using the preferred pyrolysis settings, Py-MIRS as well as DRIFTS results further indicated that soils receiving organic (e.g. farmyard manure) inputs were highly enriched in aliphatic groups, while their absence favored the accumulation of carboxyl and aromatic groups as well as polysaccharides. Py-MIRS allowed via semi-quantification of pyrolysis products a rapid monitoring of SOM bulk chemistry with a high degree of reproducibility. It was concluded that Py-MIRS represents a fast, effective and reproducible technique to characterize changes in the SOM bulk chemistry as a result of management practices. It also allows to minimize acknowledged constraints of other analytical techniques used to characterize SOM bulk chemistry such as mineral interferences and associated secondary reactions.

Two dimensional NMR spectroscopy for molecular characterization of soil organic matter: Application to boreal soils and litter

Organic soils in boreal ecosystems and peatlands represent a huge global carbon pool and their composition strongly affects soil properties. Nevertheless, the characterization of soil organic matter (SOM) molecular composition, which is essential for elucidating soil carbon processes and turnover, is not easily achieved, and further advances in the area are greatly needed. Two dimensional (2D) liquid state 1H–13C nuclear magnetic resonance (NMR) spectroscopy has been used on dimethyl sulfoxide (DMSO) extracts of SOM to achieve molecular level characterization, with signals from many identifiable molecular groups observable. Here we show that a simple and fast sample preparation allows acquisition of 2D 1H–13C NMR spectra from extracts of plant litter and organic layers in boreal ecosystems, with fast data acquisition. Our 2D NMR spectra revealed several differences in the DMSO extracts of different tree litter samples, O-horizons of forest soil, peat-forming moss (Sphagnum) and peat. The results mirror established differences between OM in soils and litter of different forest ecosystems (e.g. between deciduous and coniferous litter) but also provide indications for research to untangle previously conflicting results (e.g. cutin degradation in soil or carbohydrate degradation in peat). Thus, combination of 2D NMR methods can greatly improve analysis of litter composition and SOM composition, thereby facilitating the elucidation of their roles in biogeochemical and ecological processes that are critical for foreseeing feedback mechanisms for SOM turnover as a result of global environmental change.

示差走査熱量測定（DSC）分析による水田および田畑輪換田の土壌有機物の特徴づけ

示差走査熱量測定（DSC）分析による水田および田畑輪換田の土壌有機物の特徴づけ