Predicting Stability of Barley Straw-Derived Biochars Using Fourier Transform Infrared Spectroscopy.

In order to estimate the ability of biochar to sequester carbon as part of greenhouse gas removal technology, there is a need for rapid and accessible estimations of biochar stability. This study employs a novel method using Fourier transform infrared spectroscopy (FTIR) to predict common stability indicators, namely H:C and O:C molar ratios. Biochars derived from barley straw were produced at temperatures from 150 to 700 °C. The greatest compositional changes of the biochars occurred between 200 and 400 °C. All biochars produced at ≥400 °C achieved H:C < 0.7 and O:C < 0.4, indicative of biochars suitable for soil application. Regression models were built using FTIR data to predict H:C and O:C molar ratios. The H:C model produced a coefficient of determination (R 2) of 0.99, mean absolute percentage error (MAPE) 6.86%, and root-mean-square error (RMSE) of 0.07. The O:C model achieved the same R 2 (0.99), MAPE of 9.02%, and RMSE of 0.03. Our results demonstrate that combining FTIR data with modeling is a promising rapid and accessible method for attaining biochar stability data.

Organic Biosignature Degradation in Hydrothermal and Serpentinizing Environments: Implications for Life Detection on Icy Moons and Mars.

Evidence of liquid water is a primary indicator of habitability on the icy moons in our outer solar system as well as on terrestrial planets such as Mars. If liquid water-containing environments host life, some of its organic remains can be fossilized and preserved as organic biosignatures. However, inorganic materials may also be present and water-assisted organic-inorganic reactions can transform the organic architecture of biological remains. Our understanding of the fate of these organic remains can be assisted by experimental simulations that monitor the chemical changes that occur in microbial organic matter due to the presence of water and minerals. We performed hydrothermal experiments at temperatures between 100°C and 300°C involving lipid-rich microbes and natural serpentinite mineral mixtures generated by the subaqueous hydrothermal alteration of ultramafic rock. The products reveal what the signals of life may look like when subjected to water-organic-inorganic reactions. Straight- and branched-chain lipids in unaltered samples are joined by cyclization and aromatization products in hydrothermally altered samples. Hydrothermal reactions produce distinct products that are not present in the starting materials, including small, single-ring, heteroatomic, and aromatic compounds such as indoles and phenols. Hydrothermal reactions in the presence of serpentinite minerals lead to significant reduction of these organic structures and their replacement by diketopiperazines (DKPs) and dihydropyrazines (DHPs), which may be compounds that are distinct to organic-inorganic reactions. Given that the precursors of DKPs and DHPs are normally lost during early diagenesis, the presence of these compounds can be an indicator of coexisting recent life and hydrothermal processing in the presence of minerals. However, laboratory experiments reveal that the formation and preservation of these compounds can only occur within a distinct temperature window. Our findings are relevant to life detection missions that aim to access hydrothermal and serpentinizing environments in the subsurfaces of icy moons and Mars.

Solid-Phase Microextraction for Organic Contamination Control Throughout Assembly and Operational Phases of Space Missions.

Space missions concerned with life detection contain highly sensitive instruments for the detection of organics. Terrestrial contamination can interfere with signals of indigenous organics in samples and has the potential to cause false-positive biosignature detections, which may lead to incorrect suggestions of the presence of life elsewhere in the solar system. This study assessed the capability of solid-phase microextraction (SPME) as a method for monitoring organic contamination encountered by spacecraft hardware during assembly and operation. SPME-gas chromatography-mass spectrometry (SPME-GC-MS) analysis was performed on potential contaminant source materials, which are commonly used in spacecraft construction. The sensitivity of SPME-GC-MS to organics was assessed in the context of contaminants identified in molecular wipes taken from hardware surfaces on the ExoMars Rosalind Franklin rover. SPME was found to be effective at detecting a wide range of common organic contaminants that include aromatic hydrocarbons, aliphatic hydrocarbons, nitrogen-containing compounds, alcohols, and carbonyls. A notable example of correlation of contaminant with source material was the detection of benzenamine compounds in an epoxy adhesive analyzed by SPME-GC-MS and in the ExoMars rover surface wipe samples. The current form of SPME-GC-MS does not enable quantitative evaluation of contaminants, nor is it suitable for the detection of every group of organic molecules relevant to astrobiological contamination concerns, namely large and/or polar molecules such as amino acids. However, it nonetheless represents an effective new monitoring method for rapid, easy identification of organic contaminants commonly present on spacecraft hardware and could thus be utilized in future space missions as part of their contamination control and mitigation protocols.

Characteristics of fine and ultrafine aerosols in the London underground.

Underground railway systems are recognised spaces of increased personal pollution exposure. We studied the number-size distribution and physico-chemical characteristics of ultrafine (PM0.1), fine (PM0.1-2.5) and coarse (PM2.5-10) particles collected on a London underground platform. Particle number concentrations gradually increased throughout the day, with a maximum concentration between 18:00 h and 21:00 h (local time). There was a maximum decrease in mass for the PM2.5, PM2.5-10 and black carbon of 3.9, 4.5 and ~ 21-times, respectively, between operable (OpHrs) and non-operable (N-OpHrs) hours. Average PM10 (52 µg m-3) and PM2.5 (34 µg m-3) concentrations over the full data showed levels above the World Health Organization Air Quality Guidelines. Respiratory deposition doses of particle number and mass concentrations were calculated and found to be two- and four-times higher during OpHrs compared with N-OpHrs, reflecting events such as train arrival/departure during OpHrs. Organic compounds were composed of aromatic hydrocarbons and polycyclic aromatic hydrocarbons (PAHs) which are known to be harmful to health. Specific ratios of PAHs were identified for underground transport that may reflect an interaction between PAHs and fine particles. Scanning transmission electron microscopy (STEM) chemical maps of fine and ultrafine fractions show they are composed of Fe and O in the form of magnetite and nanosized mixtures of metals including Cr, Al, Ni and Mn. These findings, and the low air change rate (0.17 to 0.46 h-1), highlight the need to improve the ventilation conditions.

The Winchcombe meteorite, a unique and pristine witness from the outer solar system.

Direct links between carbonaceous chondrites and their parent bodies in the solar system are rare. The Winchcombe meteorite is the most accurately recorded carbonaceous chondrite fall. Its pre-atmospheric orbit and cosmic-ray exposure age confirm that it arrived on Earth shortly after ejection from a primitive asteroid. Recovered only hours after falling, the composition of the Winchcombe meteorite is largely unmodified by the terrestrial environment. It contains abundant hydrated silicates formed during fluid-rock reactions, and carbon- and nitrogen-bearing organic matter including soluble protein amino acids. The near-pristine hydrogen isotopic composition of the Winchcombe meteorite is comparable to the terrestrial hydrosphere, providing further evidence that volatile-rich carbonaceous asteroids played an important role in the origin of Earth's water.

Hydrothermal Processing of Microorganisms: Mass Spectral Signals of Degraded Biosignatures for Life Detection on Icy Moons.

Life detection missions to the outer solar system are concentrating on the icy moons of Jupiter and Saturn and their inferred subsurface oceans. Access to evidence of habitability, and possibly even life, is facilitated by the ejection of subsurface material in plumes and outgassing fissures. Orbiting spacecraft can intersect the plume material or detect past sputtered remnants of outgassed products and analyze the contents using instruments such as mass spectrometers. Hydrothermalism has been proposed for the subsurface environments of icy moons, and the organic remains of any associated life would be expected to suffer some degradation through hydrothermalism, radiolysis, or spacecraft flyby impact fragmentation. Hydrothermalism is treated here for the first time in the context of the Europa Clipper mission. To assess the influence of hydrothermalism on the ability of orbiting mass spectrometers to detect degrading signals of life, we have subjected Earth microorganisms to laboratory hydrothermal processing. The processed microorganism samples were then analyzed using gas chromatography-mass spectrometry (GC-MS), and mass spectra were generated. Certain compound classes, such as carbohydrates and proteins, are significantly altered by hydrothermal processing, resulting in small one-ring and two-ring aromatic compounds such as indoles and phenols. However, lipid fragments, such as fatty acids, retain their fidelity, and their provenance is easily recognized as biological in origin. Our data indicate that mass spectrometry measurements in the plumes of icy moons, using instruments such as the MAss Spectrometer for Planetary Exploration (MASPEX) onboard the upcoming Europa Clipper mission, can reveal the presence of life even after significant degradation by hydrothermal processing has taken place.

Mineral Matrix Effects on Pyrolysis Products of Kerogens Infer Difficulties in Determining Biological Provenance of Macromolecular Organic Matter at Mars.

Ancient martian organic matter is likely to take the form of kerogen-like recalcitrant macromolecular organic matter (MOM), existing in close association with reactive mineral surfaces, especially iron oxides. Detecting and identifying a biological origin for martian MOM will therefore be of utmost importance for life-detection efforts at Mars. We show that Type I and Type IV kerogens provide effective analogues for putative martian MOM of biological and abiological (meteoric) provenances, respectively. We analyze the pyrolytic breakdown products when these kerogens are mixed with mineral matrices highly relevant for the search for life on Mars. We demonstrate that, using traditional thermal techniques as generally used by the Sample Analysis at Mars and Mars Organic Molecule Analyser instruments, even the breakdown products of highly recalcitrant MOM are transformed during analysis in the presence of reactive mineral surfaces, particularly iron. Analytical transformation reduces the diagnostic ability of this technique, as detected transformation products of both biological and abiological MOM may be identical (low molecular weight gas phases and benzene) and indistinguishable. The severity of transformational effects increased through calcite < kaolinite < hematite < nontronite < magnetite < goethite. Due to their representation of various habitable aqueous environments and the preservation potential of organic matter by iron, it is not advisable to completely avoid iron-rich strata. We conclude that hematite-rich localities, with evidence of extensive aqueous alteration of originally reducing phases, such as the Vera Rubin Ridge, may be relatively promising targets for identifying martian biologically sourced MOM.

Transformation of Cyanobacterial Biomolecules by Iron Oxides During Flash Pyrolysis: Implications for Mars Life-Detection Missions.

Answering the question of whether life ever existed on Mars is a key goal of both NASA's and ESA's imminent Mars rover missions. The obfuscatory effects of oxidizing salts, such as perchlorates and sulfates, on organic matter during thermal decomposition analysis techniques are well established. Less well studied are the transformative effects of iron oxides and (oxy)hydroxides, which are present in great abundances in the martian regolith. We examined the products of flash pyrolysis-gas chromatography-mass spectrometry (a technique analogous to the thermal techniques employed by past, current, and future landed Mars missions) which form when the cyanobacteria Arthrospira platensis are heated in the presence of a variety of Mars-relevant iron-bearing minerals. We found that iron oxides/(oxy)hydroxides have transformative effects on the pyrolytic products of cyanobacterial biomolecules. Both the abundance and variety of molecular species detected were decreased as iron substrates transformed biomolecules, by both oxidative and reductive processes, into lower fidelity alkanes, aromatic and aryl-bonded hydrocarbons. Despite the loss of fidelity, a suite that contains mid-length alkanes and polyaromatic hydrocarbons and/or aryl-bonded molecules in iron-rich samples subjected to pyrolysis may allude to the transformation of cyanobacterially derived mid-long chain length fatty acids (particularly unsaturated fatty acids) originally present in the sample. Hematite was found to be the iron oxide with the lowest transformation potential, and because this iron oxide has a high affinity for codeposition of organic matter and preservation over geological timescales, sampling at Mars should target sediments/strata that have undergone a diagenetic history encouraging the dehydration, dihydroxylation, and oxidation of more reactive iron-bearing phases to hematite by looking for (mineralogical) evidence of the activity of oxidizing, acidic/neutral, and either hot or long-lived fluids.

Pyrolysis of Carboxylic Acids in the Presence of Iron Oxides: Implications for Life Detection on Missions to Mars.

The search for, and characterization of, organic matter on Mars is central to efforts in identifying habitable environments and detecting evidence of life in the martian surface and near surface. Iron oxides are ubiquitous in the martian regolith and are known to be associated with the deposition and preservation of organic matter in certain terrestrial environments, thus iron oxide-rich sediments are potential targets for life-detection missions. The most frequently used protocol for martian organic matter characterization (also planned for use on ExoMars) has been thermal extraction for the transfer of organic matter to gas chromatography-mass spectrometry (GC-MS) detectors. For the effective use of thermal extraction for martian samples, it is necessary to explore how potential biomarker organic molecules evolve during this process in the presence of iron oxides. We have thermally decomposed iron oxides simultaneously with (z)-octadec-9-enoic and n-octadecanoic acids and analyzed the products through pyrolysis-GC-MS. We found that the thermally driven dehydration, reduction, and recrystallization of iron oxides transformed fatty acids. Overall detectability of products greatly reduced, molecular diversity decreased, unsaturated products decreased, and aromatization increased. The severity of this effect increased as reduction potential of the iron oxide and inferred free radical formation increased. Of the iron oxides tested hematite showed the least transformative effects, followed by magnetite, goethite, then ferrihydrite. It was possible to identify the saturation state of the parent carboxylic acid at high (0.5 wt %) concentrations by the distribution of n-alkylbenzenes in the pyrolysis products. When selecting life-detection targets on Mars, localities where hematite is the dominant iron oxide could be targeted preferentially, otherwise thermal analysis of carboxylic acids, or similar biomarker molecules, will lead to enhanced polymerization, aromatization, and breakdown, which will in turn reduce the fidelity of the original biomarker, similar to changes normally observed during thermal maturation.

Portable and rapid arsenic speciation in synthetic and natural waters by an As(V)-selective chemisorbent, validated against anodic stripping voltammetry.

Inorganic arsenic speciation, i.e. the differentiation between arsenite and arsenate, is an important step for any program aiming to address the global issue of arsenic contaminated groundwater, whether for monitoring purposes or the development of new water treatment regimes. Reliable speciation by easy-to-use, portable and cost-effective analytical techniques is still challenging for both synthetic and natural waters. Here we demonstrate the first application of an As(V)-selective chemisorbent material for simple and portable speciation of arsenic using handheld syringes, enabling high sample throughput with minimal set-up costs. We first show that ImpAs efficiently removes As(V) from a variety of synthetic groundwaters with a single treatment, whilst As(III) is not retained. We then exemplify the potential of ImpAs for simple and fast speciation by determining rate constants for the photooxidation of As(III) in the presence of a TiO2 photocatalyst. Finally, we successfully speciate natural waters spiked with a mix of As(III) and As(V) in both Indian and UK groundwaters with less than 5 mg L-1 dissolved iron. Experimental results using ImpAs agreed with anodic stripping voltammetry (ASV), a benchmark portable technique, with analysis conditions optimised here for the groundwaters of South Asia. This new analytical tool is simple, portable and fast, and should find applications within the overall multi-disciplinary remediation effort that is taking place to tackle this worldwide arsenic problem.

Solid Phase Micro Extraction: Potential for Organic Contamination Control for Planetary Protection of Life-Detection Missions to the Icy Moons of the Outer Solar System.

Conclusively detecting, or ruling out the possibility of, life on the icy moons of the outer Solar System will require spacecraft missions to undergo rigorous planetary protection and contamination control procedures to achieve extremely low levels of organic terrestrial contamination. Contamination control is necessary to avoid forward contamination of the body of interest and to avoid the detection of false-positive signals, which could either mask indigenous organic chemistry of interest or cause an astrobiological false alarm. Here we test a new method for rapidly and inexpensively assessing the organic cleanliness of spaceflight hardware surfaces using solid phase micro extraction (SPME) fibers to directly swab surfaces. The results suggest that the method is both time and cost efficient. The SPME-gas chromatography-mass spectrometry (SPME-GC-MS) method is sensitive to common midweight, nonpolar contaminant compounds, for example, aliphatic and aromatic hydrocarbons, which are common contaminants in laboratory settings. While we demonstrate the potential of SPME for surface sampling, the GC-MS instrumentation restricts the SPME-GC-MS technique's sensitivity to larger polar and nonvolatile compounds. Although not used in this study, to increase the potential range of detectable compounds, SPME can also be used in conjunction with high-performance liquid chromatography/liquid chromatography-mass spectrometry systems suitable for polar analytes (Kataoka et al., 2000). Thus, our SPME method presents an opportunity to monitor organic contamination in a relatively rapid and routine way that produces information-rich data sets.

The Search for Hesperian Organic Matter on Mars: Pyrolysis Studies of Sediments Rich in Sulfur and Iron.

Jarosite on Mars is of significant geological and astrobiological interest, as it forms in acidic aqueous conditions that are potentially habitable for acidophilic organisms. Jarosite can provide environmental context and may host organic matter. The most common extraction technique used to search for organic compounds on the surface of Mars is pyrolysis. However, thermal decomposition of jarosite releases oxygen into pyrolysis ovens, which degrades organic signals. Jarosite has a close association with the iron oxyhydroxide goethite in many depositional/diagenetic environments. Hematite can form by dehydration of goethite or directly from jarosite under certain aqueous conditions. Goethite and hematite are significantly more amenable than jarosite for pyrolysis experiments employed to search for organic matter. Analysis of the mineralogy and organic chemistry of samples from a natural acidic stream revealed a diverse response for organic compounds during pyrolysis of goethite-rich layers but a poor response for jarosite-rich or mixed jarosite-goethite samples. Goethite units that are associated with jarosite, but do not contain jarosite themselves, should be targeted for organic detection pyrolysis experiments on Mars. These findings are extremely timely, as exploration targets for Mars Science Laboratory include Vera Rubin Ridge (formerly known as "Hematite Ridge"), which may have formed from goethite precursors. Key Words: Mars-Pyrolysis-Jarosite-Goethite-Hematite-Biosignatures. Astrobiology 18, 454-464.

Multiple Cosmic Sources for Meteorite Macromolecules?

The major organic component in carbonaceous meteorites is an organic macromolecular material. The Murchison macromolecular material comprises aromatic units connected by aliphatic and heteroatom-containing linkages or occluded within the wider structure. The macromolecular material source environment remains elusive. Traditionally, attempts to determine source have strived to identify a single environment. Here, we apply a highly efficient hydrogenolysis method to liberate units from the macromolecular material and use mass spectrometric techniques to determine their chemical structures and individual stable carbon isotope ratios. We confirm that the macromolecular material comprises a labile fraction with small aromatic units enriched in (13)C and a refractory fraction made up of large aromatic units depleted in (13)C. Our findings suggest that the macromolecular material may be derived from at least two separate environments. Compound-specific carbon isotope trends for aromatic compounds with carbon number may reflect mixing of the two sources. The story of the quantitatively dominant macromolecular material in meteorites appears to be made up of more than one chapter.

Heat, Aromatic Units, and Iron-Rich Phyllosilicates: A Mechanism for Making Macromolecules in the Early Solar System.

The major organic component in carbonaceous chondrites is a highly aromatic macromolecular material. Aromatic organic matter and phyllosilicates are colocated in these meteorites, and it is possible that the physical association represents a synthetic chemical relationship. To explore the potential reactions that could take place to produce the aromatic macromolecular material, we heated various simple aromatic units in the presence of montmorillonite with different exchanged cations. The majority of cation-exchanged montmorillonites tested, sodium-, aluminum-, iron-, nickel-, and cobalt-rich montmorillonites, do not produce polymerization products. By contrast, Fe(3+) cation-exchanged montmorillonite readily facilitates addition reactions between aromatic hydrocarbons. A feasible mechanism for the process is oxidative coupling, which involves a corresponding reduction of the Fe(3+) cation to its Fe(2+) counterpart. A similar reduction process for the other metal cations does not take place, highlighting the importance of iron. This simple process is one feasible mechanism for the construction of aromatic macromolecules such as those found in carbonaceous chondrites. The search for a relationship between Fe(3+)-rich phyllosilicates and aromatic organic structures (particularly dimers, trimers, and more polymerized forms) in carbonaceous chondrites would represent an effective test for constraining the role of clay catalysis in the early Solar System.

Subcritical water extraction of organic matter from sedimentary rocks.

Subcritical water extraction of organic matter containing sedimentary rocks at 300°C and 1500 psi produces extracts comparable to conventional solvent extraction. Subcritical water extraction of previously solvent extracted samples confirms that high molecular weight organic matter (kerogen) degradation is not occurring and that only low molecular weight organic matter (free compounds) are being accessed in analogy to solvent extraction procedures. The sedimentary rocks chosen for extraction span the classic geochemical organic matter types. A type I organic matter-containing sedimentary rock produces n-alkanes and isoprenoidal hydrocarbons at 300°C and 1500 psi that indicate an algal source for the organic matter. Extraction of a rock containing type II organic matter at the same temperature and pressure produces aliphatic hydrocarbons but also aromatic compounds reflecting the increased contributions from terrestrial organic matter in this sample. A type III organic matter-containing sample produces a range of non-polar and polar compounds including polycyclic aromatic hydrocarbons and oxygenated aromatic compounds at 300°C and 1500 psi reflecting a dominantly terrestrial origin for the organic materials. Although extraction at 300°C and 1500 psi produces extracts that are comparable to solvent extraction, lower temperature steps display differences related to organic solubility. The type I organic matter produces no products below 300°C and 1500 psi, reflecting its dominantly aliphatic character, while type II and type III organic matter contribute some polar components to the lower temperature steps, reflecting the chemical heterogeneity of their organic inventory. The separation of polar and non-polar organic compounds by using different temperatures provides the potential for selective extraction that may obviate the need for subsequent preparative chromatography steps. Our results indicate that subcritical water extraction can act as a suitable replacement for conventional solvent extraction of sedimentary rocks, but can also be used for any organic matter containing mineral matrix, including soils and recent sediments, and has the added benefit of tailored extraction for analytes of specific polarities.

Sulfate minerals: a problem for the detection of organic compounds on Mars?

The search for in situ organic matter on Mars involves encounters with minerals and requires an understanding of their influence on lander and rover experiments. Inorganic host materials can be helpful by aiding the preservation of organic compounds or unhelpful by causing the destruction of organic matter during thermal extraction steps. Perchlorates are recognized as confounding minerals for thermal degradation studies. On heating, perchlorates can decompose to produce oxygen, which then oxidizes organic matter. Other common minerals on Mars, such as sulfates, may also produce oxygen upon thermal decay, presenting an additional complication. Different sulfate species decompose within a large range of temperatures. We performed a series of experiments on a sample containing the ferric sulfate jarosite. The sulfate ions within jarosite break down from 500 °C. Carbon dioxide detected during heating of the sample was attributed to oxidation of organic matter. A laboratory standard of ferric sulfate hydrate released sulfur dioxide from 550 °C, and an oxygen peak was detected in the products. Calcium sulfate did not decompose below 1000 °C. Oxygen released from sulfate minerals may have already affected organic compound detection during in situ thermal experiments on Mars missions. A combination of preliminary mineralogical analyses and suitably selected pyrolysis temperatures may increase future success in the search for past or present life on Mars.

Minimising hydrogen sulphide generation during steam assisted production of heavy oil.

The majority of global petroleum is in the form of highly viscous heavy oil. Traditionally heavy oil in sands at shallow depths is accessed by large scale mining activities. Recently steam has been used to allow heavy oil extraction with greatly reduced surface disturbance. However, in situ thermal recovery processes can generate hydrogen sulphide, high levels of which are toxic to humans and corrosive to equipment. Avoiding hydrogen sulphide production is the best possible mitigation strategy. Here we use laboratory aquathermolysis to reproduce conditions that may be experienced during thermal extraction. The results indicate that hydrogen sulphide generation occurs within a specific temperature and pressure window and corresponds to chemical and physical changes in the oil. Asphaltenes are identified as the major source of sulphur. Our findings reveal that for high sulphur heavy oils, the generation of hydrogen sulphide during steam assisted thermal recovery is minimal if temperature and pressure are maintained within specific criteria. This strict pressure and temperature dependence of hydrogen sulphide release can allow access to the world's most voluminous oil deposits without generating excessive amounts of this unwanted gas product.

Contrasting wetland CH4 emission responses to simulated glacial atmospheric CO2 in temperate bogs and fens.

Wetlands were the largest source of atmospheric methane (CH(4) ) during the Last Glacial Maximum (LGM), but the sensitivity of this source to exceptionally low atmospheric CO(2) concentration ([CO(2) ]) at the time has not been examined experimentally. We tested the hypothesis that LGM atmospheric [CO(2) ] reduced CH(4) emissions as a consequence of decreased photosynthate allocation to the rhizosphere. We exposed minerotrophic fen and ombrotrophic bog peatland mesocosms to simulated LGM (c. 200 ppm) or ambient (c. 400 ppm) [CO(2) ] over 21 months (n = 8 per treatment) and measured gaseous CH(4) flux, pore water dissolved CH(4) and volatile fatty acid (VFA; an indicator of plant carbon supply to the rhizosphere) concentrations. Cumulative CH(4) flux from fen mesocosms was suppressed by 29% (P < 0.05) and rhizosphere pore water [CH(4) ] by c. 50% (P < 0.01) in the LGM [CO(2) ], variables that remained unaffected in bog mesocosms. VFA analysis indicated that changes in plant root exudates were not the driving mechanism behind these results. Our data suggest that the LGM [CO(2) ] suppression of wetland CH(4) emissions is contingent on trophic status. The heterogeneous response may be attributable to differences in species assemblage that influence the dominant CH(4) production pathway, rhizosphere supplemented photosynthesis and CH(4) oxidation.

Rapid determination of spore chemistry using thermochemolysis gas chromatography-mass spectrometry and micro-Fourier transform infrared spectroscopy.

Spore chemistry is at the centre of investigations aimed at producing a proxy record of harmful ultraviolet radiation (UV-B) through time. A biochemical proxy is essential owing to an absence of long-term (century or more) instrumental records. Spore cell material contains UV-B absorbing compounds that appear to be synthesised in variable amounts dependent on the ambient UV-B flux. To facilitate these investigations we have developed a rapid method for detecting variations in spore chemistry using combined thermochemolysis gas chromatography-mass spectrometry and micro-Fourier transform infrared spectroscopy. Our method was tested using spores obtained from five populations of the tropical lycopsid Lycopodium cernuum growing across an altitudinal gradient (650-1981 m a.s.l.) in S.E. Asia with the assumption that they experienced a range of UV-B radiation doses. Thermochemolysis and subsequent pyrolysis liberated UV-B pigments (ferulic and para-coumaric acid) from the spores. All of the aromatic compounds liberated from spores by thermochemolysis and pyrolysis were active in UV-B protection. The various functional groups associated with UV-B protecting pigments were rapidly detected by micro-FTIR and included the aromatic C[double bond, length as m-dash]C absorption band which was exclusive to the pigments. We show increases in micro-FTIR aromatic absorption (1510 cm(-1)) with altitude that may reflect a chemical response to higher UV-B flux. Our results indicate that rapid chemical analyses of historical spore samples could provide a record ideally suited to investigations of a proxy for stratospheric O3 layer variability and UV-B flux over historical (century to millennia) timescales.