Ultra-high resolution mass spectrometry of physical speciation patterns of organic matter in fire-affected soils.

Fire is one of the most important modulating factors of the environment and the forest inducing chemical and biological changes on the most reactive soil component, the soil organic matter (SOM). Assuming the complex composition of the SOM, we used an ultra-high resolution mass spectrometry analysis technique to assess the chemical composition and fire-induced alterations in soil particle size fractions (coarse and fine) from a sandy soil in a Mediterranean oak forest at Doñana National Park (Southwest Spain). Electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICRMS) showed that the coarse fraction of soils not affected by fires consisted mainly of polyphenolic compounds consistent with little-transformed SOM and fresh biomass, whereas the fine fraction was enriched in protein and lipid like homologues suggesting microbially reworked SOM. In fire-affected SOM, the coarse fraction contained a high proportion of aromatic compounds, consistent with inputs of charred litter or in situ chemical transformation of the SOM. Analysis of the fine fraction revealed two differentiated chemical families pointing to the existence of two carbon pools; a native microbial-derived moiety composed of lipids and polypeptide compounds, and a secondary, pyrogenic or thermally-altered moiety rich in aromatic compounds. This work represents the first application of ultra-high resolution mass spectrometry to study the chemical composition of SOM in different particle size fractions.

Wildfire effects on lipid composition and hydrophobicity of bulk soil and soil size fractions under Quercus suber cover (SW-Spain).

Soil water repellency (hydrophobicity) prevents water from wetting or infiltrating soils, triggering changes in the ecosystems. Fire may develop, enhance or destroy hydrophobicity in previously wettable or water-repellent soils. Soil water repellency is mostly influenced by the quality and quantity of soil organic matter, particularly the lipid fraction. Here we report the results of a study on the effect of fire on the distribution of soil lipids and their role in the hydrophobicity grade of six particle size fractions (2-1, 1-0.5, 0.5-0.25, 0.25-0.1, 0.1-0.05 and <0.05mm) of an Arenosol under Quercus suber canopy at the Doñana National Park (SW-Spain). Hydrophobicity was determined using water drop penetration time test. Field emission scanning electron microscopy (FESEM) was used to assess the presence and morphology of the inorganic and organic soil components in the particle size fractions. Soil lipids were Soxhlet extracted with a dichloromethane-methanol mixture. Fatty acids (FAs) and neutral lipids were separated, derivatized, identified and quantified by gas chromatography/mass spectrometry and gas chromatography/flame ionization detection. The hydrophobicity values of soil samples and fractions were statistically different (P < 0.05), for both, the unburnt and burnt soils, and particle size fractions. All samples displayed a similar distribution of FAs, straight-chain saturated acids in the C14-C32 range, and neutral lipids (n-alkan-1-ols, n-alkanes), only differing in their relative abundances. Among possible biogeochemical mechanisms responsible for the changes in soil lipids, the observed depletion of long chain FAs (C≥24) in the coarse fraction is best explained by thermal cracking caused by the heat of the fire. The enrichment of long chain FAs observed in other fractions suggests possible exogenous additions of charred, lipid-rich, material, like cork suberin or other plant-derived macromolecules (cutins). Principal component analysis was used to study the relationships between hydrophobicity with soil organic matter and its different components. Extractable organic matter (EOM) and specifically long chain FAs content were positively correlated to soil hydrophobicity. Therefore, the latter could be used as biomarkers surrogated to hydrophobicity in sandy soils.

Compound-specific stable carbon isotopic signature of carbohydrate pyrolysis products from C3 and C4 plants.

BACKGROUND: Pyrolysis-compound specific isotopic analysis (Py-CSIA: Py-GC-(FID)-C-IRMS) is a relatively novel technique that allows on-line quantification of stable isotope proportions in chromatographically separated products released by pyrolysis. Validation of the Py-CSIA technique is compulsory for molecular traceability in basic and applied research. In this work, commercial sucrose from C4 (sugarcane) and C3 (sugarbeet) photosystem plants and admixtures were studied using analytical pyrolysis (Py-GC/MS), bulk δ(13)C IRMS and δ(13)C Py-CSIA. RESULTS: Major pyrolysis compounds were furfural (F), furfural-5-hydroxymethyl (HMF) and levoglucosan (LV). Bulk and main pyrolysis compound δ(13)C (‰) values were dependent on plant origin: C3 (F, -24.65 ± 0.89; HMF, -22.07 ± 0.41‰; LV, -21.74 ± 0.17‰) and C4 (F, -14.35 ± 0.89‰; HMF, -11.22 ± 0.54‰; LV, -11.44 ± 1.26‰). Significant regressions were obtained for δ(13)C of bulk and pyrolysis compounds in C3 and C4 admixtures. Furfural (F) was found (13)C depleted with respect to bulk and HMF and LV, indicating the incorporation of the light carbon atom in position 6 of carbohydrates in the furan ring after pyrolysis. CONCLUSION: This is the first detailed report on the δ(13)C signature of major pyrolytically generated carbohydrate-derived molecules. The information provided by Py-CSIA is valuable for identifying source marker compounds of use in food science/fraud detection or in environmental research.

Molecular characterisation of a bio-based active packaging containing Origanum vulgare L. essential oil using pyrolysis gas chromatography-mass spectrometry.

BACKGROUND: Environmental, economic and safety challenges motivate shift towards safer materials for food packaging. New bioactive packaging techniques, i.e. addition of essential plant oils (EOs), are gaining attention by creating barriers to protect products from spoilage. Analytical pyrolysis gas chromatography-mass spectrometry (Py-GC-MS) was used to fingerprint a bioactive polylactic acid (PLA) with polybutylene succinate (PBS) (950 g kg(-1) :50 g kg(-1) ) film extruded with variable quantities (0, 20, 50 and 100 g kg(-1) ) of Origanum vulgare EO. RESULTS: Main PLA:PBS pyrolysis products were lactide enantiomers and monomer units from the major PLA fraction and succinic acid anhydride from the PBS fraction. Oregano EO pyrolysis released cymene, terpinene and thymol/carvacrol peaks as diagnostic peaks for EO. In fact, linear correlation coefficients better than 0.950R(2) value (P < 0.001) were found between the chromatographic area of the diagnostic peaks and the amount of oregano EO in the bioplastic. CONCLUSION: The pyrolytic behaviour of a bio-based active package polymer including EO is studied in detail. Identified diagnostic compounds provide a tool to monitor the quantity of EO incorporated into the PLA:PBS polymeric matrix. Analytical pyrolysis is proposed as a rapid technique for the identification and quantification of additives within bio-based plastic matrices. © 2015 Society of Chemical Industry.

Pyrolysis-gas chromatography/mass spectrometry identification of distinctive structures providing humic character to organic materials.

Flash pyrolysis-gas chromatography/mass spectrometry(Py-GC/MS) was used to study the structural transformations of humic fractions formed as a result of composting processes of diverse organic materials (solid wastes of wineries, solid olive-mill wastes, domestic wastes, ovine manures plus straw, and mixtures of animal manures). Sodium hydroxide-extracted total humic-like extracts (THE; humic plus fulvic acids) from the composted and the initial noncomposted wastes and several reference humic and fulvic acids from soils were analyzed. These results were compared with results from previous studies using 13C-cross polarization magic angle spinning-nuclear magnetic resonance (NMR), UV-visible, and fluorescence emission spectroscopies. Alkylbenzenes and alkylphenols predominate in the pyrograms of the soil humic acids, whereas the fulvic acids showed higher contents of phenolic and polysaccharide-derived compounds. The pyrolysates of THE from the composted samples showed an increase in aromatic and nitrogenated structures and a decrease in polysaccharide-derived compounds. The aromatic contents as determined by Py-GC/MS and 13C-NMR were well correlated in the reference humic substances and THE from composted materials (r = 0.99 and 0.94, respectively; P < 0.01) but not in the case of THE from noncomposted materials, probably due to an aliphatic enhancement in the pyrolysates of these samples and other secondary reactions. The content in alkylbenzenes was consistent with the variations found previously for several UV-visible and fluorescence indexes as a function of the degree of humification, suggesting their involvement in structures that are a characteristic feature of the formation and evolution of humic substances.

Discerning natural and anthropogenic organic matter inputs to salt marsh sediments of Ria Formosa lagoon (South Portugal).

Sedimentary organic matter (OM) origin and molecular composition provide useful information to understand carbon cycling in coastal wetlands. Core sediments from threors' Contributionse transects along Ria Formosa lagoon intertidal zone were analysed using analytical pyrolysis (Py-GC/MS) to determine composition, distribution and origin of sedimentary OM. The distribution of alkyl compounds (alkanes, alkanoic acids and alkan-2-ones), polycyclic aromatic hydrocarbons (PAHs), lignin-derived methoxyphenols, linear alkylbenzenes (LABs), steranes and hopanes indicated OM inputs to the intertidal environment from natural-autochthonous and allochthonous-as well as anthropogenic. Several n-alkane geochemical indices used to assess the distribution of main OM sources (terrestrial and marine) in the sediments indicate that algal and aquatic macrophyte derived OM inputs dominated over terrigenous plant sources. The lignin-derived methoxyphenol assemblage, dominated by vinylguaiacol and vinylsyringol derivatives in all sediments, points to large OM contribution from higher plants. The spatial distributions of PAHs (polyaromatic hydrocarbons) showed that most pollution sources were mixed sources including both pyrogenic and petrogenic. Low carbon preference indexes (CPI > 1) for n-alkanes, the presence of UCM (unresolved complex mixture) and the distribution of hopanes (C29-C36) and steranes (C27-C29) suggested localized petroleum-derived hydrocarbon inputs to the core sediments. Series of LABs were found in most sediment samples also pointing to domestic sewage anthropogenic contributions to the sediment OM.

Fire effects on C and H isotopic composition in plant biomass and soil: Bulk and particle size fractions.

This work studies carbon (C) and hydrogen (H) isotope composition of plant biomass and soil organic matter (SOM) in an attempt to assess both, changes exerted by fire and possible inputs of charred materials to the soil after a wildfire. Isotope composition of bulk soil, soil particle size fractions and biomass of the dominant standing vegetation in the area (Quercus suber) from Doñana National Park (SW-Spain) were studied by isotope ratio mass spectrometry (IRMS). SOM C isotope composition indicates the occurrence of two SOM pools with different degree of alteration. Coarse soil fractions (>0.5 mm) were found 13C depleted with δ13C values close to those in leaf biomass, pointing to a predominance of poorly transformed SOM. Conversely, fine fractions (<0.1 mm) were found enriched in 13C as corresponds to a more humified SOM. The fire produced no changes in this trend, although a consistent 13C enrichment (c. 1‰) was observed in all soil fractions with decreasing size. Concerning H isotopes, the coarse fractions (>0.5 mm) displayed significant lower δ2H values than the intermediate and fine ones (<0.5 mm), again similar to those in leaf biomass (c. -80‰), whereas the fine fractions were found deuterium (2H)-enriched with significant higher δ2H values (c. 50‰), suggesting physical speciation of H depending on soil particle size. The fire produced a significant 2H depletion (Δ2H c. -10‰) in the finer fractions (<0.1 mm). The study of stable isotope analysis added new information and complements the results obtained by other proxies to better understand the effect of fire on SOM.

Physico-chemical and microbial perturbations of Andalusian pine forest soils following a wildfire.

Wildfires are a recurrent disturbance in Mediterranean forests, triggered by high fuel load, high environmental temperature and low humidity. Although, human intervention is behind the initiation of most fire episodes, the situation is likely to worsen in the future due to the effects of climate change in the Mediterranean "hot-spot". Here we study chemical, physical and microbial characteristics of burnt soils from two well differentiated sites at Sierra de Cazorla, Segura and Las Villas Natural Park, Andalusia, (Spain) affected and unaffected by a wildfire, and followed their evolution for three years. The soils affected by a severe surface burn showed a significant increase in organic matter after 3years from the fire. Viable bacteria and fungi also increased, especially 2-3years post-burning. Substrate induced respiration (SIR) also increased significantly in burnt soil from site 1 (rendzina on carbonate) while a significant decrease was observed in the burnt soils sampled from site 2 (calcic luvisols) in samples taken one month after the wildfire. A recovery in both SIR and organic matter was observed after 2 and 3years. Of seven soil enzymes studied, only phosphatase activity was significantly higher in most burnt soils over the three years. Analysis of bacterial community diversity using clone libraries showed a recovery in the number of phyla in burnt soils after 2 and 3years in both sites, with an increase in Proteobacteria and Firmicutes and a decrease in Acidobacteria phyla. For Bacteroidetes, the percentages were lower in most burnt samples. This study reveals that if wildfire increases the organic matter availability, then the microbial community responds with increased activity and biomass production. Although fire exerts an initial impact on the soil bacterial community, its structure and functional profile soon recovers (after 2-3years) contributing to soil recovery.

The diversity of methoxyphenols released by pyrolysis-gas chromatography as predictor of soil carbon storage.

The variable extent to which environmental factors are involved in soil carbon storage is currently a subject of controversy. In fact, justifying why some soils accumulate more organic matter than others is not trivial. Some abiotic factors such as organo-mineral associations have classically been invoked as the main drivers for soil C stabilization. However, in this research indirect evidences based on correlations between soil C storage and compositional descriptors of the soil organic matter are presented. It is assumed that the intrinsic structure of soil organic matter should have a bearing in the soil carbon storage. This is examined here by focusing on the methoxyphenols released by direct pyrolysis from a wide variety of topsoil samples from continental Mediterranean ecosystems from Spain with different properties and carbon content. Methoxyphenols are typical signature compounds presumptively informing on the occurrence and degree of alteration of lignin in soils. The methoxyphenol assemblages (12 major guaiacyl- and syringyl-type compounds) were analyzed by pyrolysis-gas chromatography-mass spectrometry. The Shannon-Wiener diversity index was chosen to describe the complexity of this phenolic signature. A series of exploratory statistical analyses (simple regression, partial least squares regression, multidimensional scaling) were applied to analyze the relationships existing between chemical and spectroscopic characteristics and the carbon content in the soils. These treatments coincided in pointing out that significant correlations exist between the progressive molecular diversity of the methoxyphenol assemblages and the concentration of organic carbon stored in the corresponding soils. This potential of the diversity in the phenolic signature as a surrogate index of the carbon storage in soils is tentatively interpreted as the accumulation of plant macromolecules altered into microbially reworked structures not readily recognized by soil enzymes. From a quantitative viewpoint, the partial least squares regression models exclusively based on total abundances of the 12 major methoxyphenols were especially successful in forecasting soil carbon storage.

Analysis of carbon and nitrogen forms in soil fractions after the addition of 15N-compost by 13C and 15N nuclear magnetic resonance.

A quantitative laboratory assessment of the different C and N forms in soil humus fractions was carried out by incubation of a mineral substrate after the addition of (15)N-labeled compost. The experimental design included (i) preparation of the (15)N-labeled organic matter (city refuse compost, 640 g kg(-1) wheat straw and K(15)NO(3) composted for 80 days), (ii) a further 80 day incubation of a mixture of the labeled compost with a mineral soil (32 g kg(-1)), (iii) measurement of stable isotope ratios, and (iv) isolation and structural comparison by (13)C and (15)N cross-polarization, magic-angle spinning nuclear magnetic resonance (NMR) of different organic fractions, i.e., soluble, colloidal (humic and fulvic type), and particulate (free organic matter and humin), from both the compost and the compost-treated soil. The results showed that the amide forms dominated in all of the newly formed N compounds, but an increased amount of alkali insoluble organic fractions was observed after incubation of the soil. The analysis of the insoluble, particulate fractions shows that nonextractable amides constitute the major pool of newly formed N compounds. The particulate soil fraction isolated by flotation in CHBr(3)-MeOH contained 16.8% of the total soil N and 26% of the (15)N. The (13)C NMR spectra showed that the fulvic acid-like fraction (7.6% of the soil N, 8.8% of (15)N) consisted almost completely of a C=O-containing carbohydrate material, whereas the humic acid-like fraction (20.3% of the total soil N, 8.6% of (15)N) resembled an oxidized lignoproteic fraction containing the most significant aromatic domain. The water soluble fraction was, in both soil and compost, the one with the highest isotopic abundance of (15)N (96%), but the (15)N NMR spectrum revealed minor amounts of soluble mineral N in this fraction and the remainder consisting of amide compounds.

The effect of fire on soil organic matter--a review.

The extent of the soil organic carbon pool doubles that present in the atmosphere and is about two to three times greater than that accumulated in living organisms in all Earth's terrestrial ecosystems. In such a scenario, one of the several ecological and environmental impacts of fires is that biomass burning is a significant source of greenhouse gases responsible for global warming. Nevertheless, the oxidation of biomass is usually incomplete and a range of pyrolysis compounds and particulate organic matter (OM) in aerosols are produced simultaneously to the thermal modification of pre-existing C forms in soil. These changes lead to the evolution of the OM to "pyromorphic humus", composed by rearranged macromolecular substances of weak colloidal properties and an enhanced resistance against chemical and biological degradation. Hence the occurrence of fires in both undisturbed and agricultural ecosystems may produce long-lasting effects on soils' OM composition and dynamics. Due to the large extent of the C pool in soils, small deviations in the different C forms may also have a significant effect in the global C balance and consequently on climate change. This paper reviews the effect of forest fires on the quantity and quality of soils' OM. It is focused mainly on the most stable pool of soil C; i.e., that having a large residence time, composed of free lipids, colloidal fractions, including humic acids (HA) and fulvic acids (FA), and other resilient forms. The main transformations exerted by fire on soil humus include the accumulation of new particulate C forms highly resistant to oxidation and biological degradation including the so-called "black carbon" (BC). Controversial environmental implications of such processes, specifically in the stabilisation of C in soil and their bearing on the global C cycle are discussed.

Complementary multianalytical approach to study the distinctive structural features of the main humic fractions in solution: gray humic acid, brown humic acid, and fulvic acid.

Previous studies have indicated that the main fractions of humic substances (HS), gray humic acid (GHA), brown humic acid (BHA), and fulvic acid (FA), present different molecular patterns in water solution that are probably associated with specific structural features. However, the techniques used in these previous studies did not permit clarification of the principal qualitative characteristics of these structures. To study more in depth this subject several GHA, BHA, and FA have been analyzed through the complementary use of UV-visible and FTIR spectroscopy, (13)C NMR, thermogravimetry, and pyrolysis GC-MS. The results indicate that the studied humic fractions have different and distinctive structural features. Thus, large and nonpolar structural units (paraffins, olefins, terpenes) and aliphatic structures seem to accumulate in the gray fraction, whereas the smallest and more polar (furfural, phenols) and simpler structural units (sugar- and amino acid-related structures) are present in the fulvic one. BHA has a higher content in polycyclic aromatic moieties, S-containing compounds and aromatic structures, thus suggesting the presence of more condensed aromatic rings. Likewise, differences in both the presence of polar groups and the apparent molecular size explain the pattern of solubility as a function of pH and ionic strength (I) that defines each HS fraction. These results also indicate that the structural differences among the HS fractions are not only quantitative (the presence of the same type of structures differing in size and the concentration of functional groups) but also qualitative, because each fraction presented different and distinctive structural domains. These structural domains explain the molecular patterns associated with each HS fraction. Thus, the presence of smaller and more O-functionalized structural units including aromatic domains in FA explain their tendency to form molecular aggregates (hydrogen bridges, metal bridges, and hydrophobic interactions) in solution. This fact could also explain the presence of molecular aggregates in BHA, although to a lesser extent than in FA. Finally, the dominant aliphatic and less functionalized character of GHA may justify its lower tendency to form aggregates in solution at neutral and alkaline pH. Likewise, the results also indicate that the different structural domains associated with these fractions may be the consequence of diverse biosynthetic pathways involving different precursors.

Soil health -- a new challenge for microbiologists and chemists.

Soil health refers to the biological, chemical, and physical features of soil that are essential to long-term, sustainable agricultural productivity with minimal environmental impact. Thus, soil health provides an overall picture of soil functionality. Although it cannot be measured directly, soil health can be inferred by measuring specific soil properties (e.g. organic matter content) and by observing soil status (e.g. fertility). There is also increased interest in studying soil microorganisms in their particular environments, as microbial diversity is intimately related to soil structure and function. One of the key objectives in determining soil health is to acquire indicators that can be used to evaluate the soil's current status and hence to develop sustainable agricultural systems. In this regard, significant progress has been made over the last few years in the development of specific biomarkers and macromolecular probes, enabling rapid and reliable measurements of soil microbial communities. In addition, modern molecular biological techniques, such as fluorescence in situ hybridization (FISH), reverse transcriptase polymerase chain reaction (RT-PCR), denaturing gradient gel electrophoresis (DGGE), and terminal restriction fragment length polymorphism (T-RFLP), have facilitated the analysis of microbial biodiversity and activity, whereas the application of modern analytical techniques, such as nuclear magnetic resonance (NMR) and pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS), have provided data on soil chemistry. The combination of these two approaches offers promise in determining soil health status.

Soil health - a new challenge for microbiologists and chemists / La salud del suelo - un nuevo reto para microbiólogos y químicos

Soil health refers to the biological, chemical, and physical features of soil that are essential to long-term, sustainable agricultural productivity with minimal environmental impact. Thus, soil health provides an overall picture of soil functionality. Although it cannot be measured directly, soil health can be inferred by measuring specific soil properties (e.g. organic matter content) and by observing soil status (e.g. fertility). There is also increased interest in studying soil microorganisms in their particular environments, as microbial diversity is intimately related to soil structure and function. One of the key objectives in determining soil health is to acquire indicators that can be used to evaluate the soil’s current status and hence to develop sustainable agricultural systems. In this regard, significant progress has been made over the last few years in the development of specific biomarkers and macromolecular probes, enabling rapid and reliable measurements of soil microbial communities. In addition, modern molecular biological techniques, such as fluorescence in situ hybridization (FISH), reverse transcriptase polymerase chain reaction (RT-PCR), denaturing gradient gel electrophoresis (DGGE), and terminal restriction fragment length polymorphism (T-RFLP), have facilitated the analysis of microbial biodiversity and activity, whereas the application of modern analytical techniques, such as nuclear magnetic resonance (NMR) and pyrolysis gas chromatography-mass spectrometry (Py-GC-MS), have provided data on soil chemistry. The combination of these two approaches offers promise in determinings oil health status (AU)

El concepto de salud del suelo se relaciona con las características biológicas, químicas y físicas que son esenciales para una productividad agrícola sostenible a largo plazo con un mínimo impacto ambiental. La salud del suelo es el más fiel reflejo de su funcionalidad. Aunque no puede medirse directamente, se puede inferir a partir de la determinación de propiedades específicas del mismo suelo (p.e. el contenido en materia orgánica) y por la observación de su estado (p.e. la fertilidad). El interés por el estudio de los microorganismos del suelo en su propio medio está aumentando, puesto que la diversidad microbiana está estrechamente relacionada con la estructura y función del suelo. Un objetivo clave para determinar la salud de un suelo es la disponibilidad de indicadores factibles de ser utilizados en la evaluación de su estado y, a partir de aquí, implementar sistemas de agricultura sostenible. El desarrollo de biomarcadores específicos y sondas macromoleculares ha evolucionado considerablemente en los últimos años, lo cual ha permitido obtener medidas fiables y rápidas de las comunidades microbianas del suelo. Por otro lado, las modernas técnicas biomoleculares más recientes (p.e. la hibridación in situ mediante fluorescencia [FISH], la reacción en cadena de la polimerasa mediante transcriptasa inversa [RT-PCR], la electroforesis en gel con gradiente desnaturalizante [DGGE]y el análisis del polimorfismo en la longitud de los fragmentos de restricción terminales [T-RFLP]) permiten analizar la biodiversidad y actividad microbianas, mientras que la aplicación de técnicas analíticas modernas (p.e. resonancia magnética nuclear [NMR], pirolisis-cromatografía de gases-espectrometría de masas [Py-GC-MS]) proporciona datos sobre la química del suelo. La combinación de estas dos aproximaciones metodológicas ofrece buenas perspectivas en la determinación del estado de salud del suelo (AU)