Carbon Isotope Determinations Research Articles

Identifying Putative Subsurface Microbial Drivers of Methane Flux on Earth and Mars

On Earth microorganisms are critical drivers of the methane cycle, both producing and consuming methane (Boetius et al. 2000, Knittel and Boetius 2009, Orphan et al. 2001). Molecular and isotopic-based investigations of archaeal-bacterial consortia catalyzing the anaerobic oxidation of methane (AOM) in marine methane seeps identified the pivotal role of these microorganisms in mitigating the release of methane into the atmosphere (Knittel and Boetius 2009, Orphan et al. 2001). In the marine environment, AOM is predominantly carried out by closely associated consortia of methanotrophic archaea (ANME) and sulfate reducing bacteria (SRB) coupling methane oxidation to sulfate reduction in the absence of oxygen. Wolf Spring (WS), Axel Heiberg Island, Nunavut is a hypersaline cold spring methane seep and the only known terrestrial permafrost hosted methane seep known to host ANME-1 archaea associated with AOM (Niederberger et al. 2010, Magnuson et al. 2022). Wolf Spring is an unparalleled analogue for putative subsurface brines and sites of methane release on Mars. Enigmatic observations of methane in the near-surface Martian atmosphere remain a tantalizing potential biosignature. The combination of field site characterization, microbial microcosm experiments, and in situ methane monitoring represents a coordinated interdisciplinary effort to identify methane driven microbial metabolisms not only critical to understanding methane flux in the Arctic, but also as possible drivers to the methane cycle on Mars. Detailed microbial characterization of these springs has identified a chemotrophic community dominated by sulfur cycling (Altshuler et al. 2022, Niederberger et al. 2010). To date, microbial and geochemical characterization has been carried out on sediment samples to a few centimeters depth. This study expands on these initial studies, with the successful collection and analysis of deeper sediment cores at WS focusing on AOM activity to better understand the microorganisms involved and the methane cycling capacity at depth. Two decades of observing methane on Mars (Mumma et al. 2009) have generated data indicative of a dynamic, geochemical system characterized by a profile similar to the release of methane from seeps on Earth (Etiope and Oehler 2019) producing both distinct pulses known as plumes and slow background seepage. These observations suggest as of yet unknown geochemical and potentially geobiological methane sources and sinks. While methane can be produced abiotically (Etiope and Lollar 2013), on Earth most methane is biogenic. Determining the biogenicity of CH4 is non-trivial and requires a correlated approach including determination of carbon isotopes. In terrestrial systems, biogenic CH4 is 13C depleted. To characterize methane sources and sinks on Mars, near surface measurements at a frequency not possible with existing instrumentation are required. We are currently developing off-axis integrated cavity-enhanced output (OA-ICOS) spectrometry as a portable trace gas analyzer capable of obtaining high frequency measurements of methane at the sub-ppb level (Sapers et al. 2021). Optimizing OA-ICOS trace methane measurements at WS will help refine sensitivity and measurement cadence in a Mars-like environment as well as providing new remote methane monitoring capabilities for Arctic methane emissions. We are currently developing in situ 12CH4:13CH4 capabilities using OA-ICOS technology. The importance of δ 13C as a biosignature is summarized in Fig. 1.

Reassessing the origin of lentil cultivation in the Pre-Pottery Neolithic of Southwest Asia: new evidence from carbon isotope analysis at Gusir Höyük

Lentils represent the most common pulse crop found in Southwest Asian Neolithic archaeobotanical assemblages. Using carbon isotope determinations and palaeoclimatic modelling this paper investigates variability and change through time in lentil growth habitats at Gusir Höyük, a Pre-Pottery Neolithic habitation site in the Upper Tigris basin of southeast Anatolia dated to the 12th and 11th millennia cal bp. The elevated lentil carbon isotope values from samples dated to ~ 10,500–10,300 cal bp possibly points to greater water availability and the potential onset of lentil cultivation at Gusir Höyük during the Early Pre-Pottery Neolithic B. The paper revisits previous hypotheses about the process and timing of initial lentil domestication in the region through a re-evaluation of materials available from other Neolithic sites in SW Asia. Its findings highlight the important role of wild lentil populations growing in higher-rainfall localities, which may have responded more readily to increasing water availability in the context of early cultivation. It also explores the potential offered by isotopic approaches for detecting early cultivation, highlighting the need for future studies on pulse and cereal crop progenitors in SW Asia and beyond.

First evidence and potential sources of novel brominated flame retardants and BDE 209 in the deepest ocean

Organic anthropogenic pollutants reach even the deepest parts of the oceans, i.e., the hadal trenches. We here presented the concentrations, influencing factors, and potential sources of polybrominated diphenyl ethers (PBDEs) and novel brominated flame retardants (NBFRs) in hadal sediments and amphipods from the Mariana, Mussau and New Britain trenches. Results showed that BDE 209 was the dominant PBDEs congener and DBDPE was the dominant NBFRs. No significant correlation was found between TOC contents and PBDEs or NBFRs levels in sediment. Lipid content and body length were the potential important factors affecting variation in pollutant concentrations in the carapace & muscle of amphipods, while the pollution levels of viscera were mainly affected by the sex and lipid content. PBDEs and NBFRs might reach trench surface seawater through long-range atmospheric transport and oceans currents but with little contribution from the Great Pacific Garbage Patch. Determination of carbon and nitrogen isotopes indicated that the pollutants were transported and accumulated in amphipods and sediment via different pathways. PBDEs and NBFRs in the hadal sediments were generally transported via the settling of sediment particles of either marine or terrigenous origin whereas in amphipods they accumulated via feeding on animal carrion through the food web. This is the first study reporting on BDE 209 and NBFR contaminations in hadal settings and provide new insight on influencing factors and sources of PBDEs and NBFRs in the deepest oceans.

Investment of needle nitrogen to photosynthesis controls the nonlinear productivity response of young Chinese fir trees to nitrogen deposition

Plant carbon (C) assimilation is expected to nonlinearly increase with continuously increasing nitrogen (N) deposition, causing a N saturation threshold for productivity. However, the response of plant productivity to N deposition rates and further the N saturation threshold still await comprehensive quantization for forest ecosystem. Here, we tested the effect of N addition on aboveground net primary productivity (ANPP) of three-year old Chinese fir (Cunninghamia lanceolata) trees by adding N at 0, 5.6, 11.2, 22.4, and 44.8 g N m−2 yr−1 for 2.5 years. The N saturation threshold was estimated based on a quadratic-plus-plateau model. Results showed that ANPP transitioned from an increasing stage with increasing N addition rate to a plateaued stage at an N rate of 16.3 g N m−2 yr−1. The response of ANPP to N addition rates was well explained by the net photosynthetic rates of needles. Results from the dual isotope measurement [simultaneous determination of needle stable carbon (δ13C) and oxygen (δ18O) isotopes] indicated that the photosynthetic capacity, rather than the stomatal conductance, mediated the response of photosynthesis and ANPP of the young Chinese fir trees to N addition. Accordingly, the amount of needle N partitioning to water-soluble fraction, which is associated with the photosynthetic capacity, also responded to N enrichment with a nonlinear increase. Our study will contribute to a more accurate prediction on the influence of N deposition on C cycles in Chinese fir plantations.

Determination of carbon isotopes in carbonates (calcite, dolomite, magnesite, and siderite) by femtosecond laser ablation multi-collector ICP-MS

This study reports a method for in situ determination of stable carbon (δ13C‰) isotope compositions for various carbonate matrices including calcite, dolomite, magnesite and siderite by fs-LA-MC-ICP-MS with a precision of 0.37–0.68‰.

Diagenesis and the Conditions of Deposition of the Middle Jurassic Siderite Rocks from the Northern Margin of the Holy Cross Mountains (Poland)

The aim of the present study is to reconstruct sedimentary conditions of Middle Jurassic rocks that contain siderites to identify the mineral composition of the inserbeds and to recognize the origin of the siderite. Thin inserbeds of siderite rocks occur most frequently within Bajocian siliciclastic deposits and, more rarely, Aalenian and Bathonian. The research material comes from 11 boreholes located in the north and northeastern margins of the Holy Cross Mountains. The research methods included sedimentological analyses, and studies in polarizing and scanning electron microscopes, staining of carbonates, cathodoluminescence, X-ray structural analysis, and stable carbon and oxygen isotopic determinations were used. Middle Jurassic sideritic rocks are most often represented by clayey siderites, which also include muddy and sandy varieties and siderite sandstones. There are also local occurrences of coquinas, claystones, mudstones, and siderite conglomerates. The main component of sideritic rocks is sideroplesite. Berthierine, pistomesite, calcite, and ankerite are important components, too. The action of diagenetic processes of cementation, compaction, replacement, and alteration within the Middle Jurassic deposits was most intense during the eo- and mesodiagenesis. The sedimentological analysis showed that most of the studied siderites were formed in a low-oxygenated marine environment, mainly in the transition zone between the normal and storm wave bases and in the lower and middle shoreface zones. The results of the petrographic, mineralogical, and geochemical studies indicated the origin of the sideritic rocks mainly in the marine environment, with the participation of meteoric water. There were slight differences in the chemical composition of sideroplesite depending on the environment it crystallized in. There was no correlation between the values of the carbon isotope determinations in the sideroplesite and the environmental conditions of its crystallization. Slight differences were visible in the case of the average values of δ18O in the sideroplesite.

Deep CO2 release revealed by stable isotope and diffuse degassing surveys at Vulcano (Aeolian Islands) in 2015–2018

The partitioning of carbon dioxide (CO2) released by soils at Vulcano Island (Aeolian Islands, Italy) was performed by combining the CO2 flux and the carbon isotope measurements. Based on this method, the amount of CO2 of volcanic origin was quantified six times during the period 2015–2018. The data analysis allowed us to establish the correlation between CO2 soil degassing and changes in the contribution of volcanic fluids. Carbon isotope determinations were performed in situ to enhance the coverage of data collection in space and time. These data were combined with both the CO2 contents in the ground gases and the soil CO2 flux. The amount of volcanic CO2 was distinguished from that of biogenic origin by implementing a three-component mixing model. The results of this study indicate that the increase in CO2 output in September 2018 reflects the increase in volcanic gas emissions. The measurement method and analysis presented in this work are sufficiently general to be applicable to the monitoring programs of active volcanoes.

Bio-geochemical evolution and critical element mineralization in the Cretaceous-Cenozoic coals from the southern Far East Russia and northeastern China

A total of 23 coal and two carbonaceous shale samples of the Early Cretaceous-Miocene, collected from different basins in the Primorye and Sakhalin regions of Russia, and the Hunchun basin of northeastern China, were used in this study for determination of major oxides (by AES and XRF), trace elements (by ICP-MS), sulfur, carbon, carbon isotopes, and major components of extractable organic matter (wet-chemical and gas-chromatographic analyses). Interpretation of the obtained data was intended to elucidate geodynamic and other geological processes controlling bio-geochemical compositions and critical element mineralization of the studied coals. The data suggest that inorganic matter in the studied coals was derived mainly from the Paleozoic granites and the Cenozoic felsic and intermediate tuffs. The Early Eocene and, in particular, Oligocene epochs represent the two major mineralization events in the regions, resulting in enrichment of Ge, W, Be, Mo, Sb, Sr, Ba, REY (rare earth elements and Y), Th, and U in the coals. The two events were related respectively to the Indo-Eurasian collision followed by plate reorganization in the Pacific and the opening of the Sea of Japan. The geodynamic changes were probably associated with mantle-derived volcanic activity and intensive degassing, which enhanced the capability of surficial and ground waters to leach and redistribute trace metals. Organic matter played a significant role in the system of mineralization, starting from weathering of source rocks and ending at the coal diagenetic stage. Organic acids significantly accelerated leaching and transportation of the metals, while they, together with sulfate-reducing bacteria, kerogen, alcohols + ketones, and n-alkanols in particular, played an important role in metal precipitation, mainly through reactions of cation exchange and adsorption.

Impact of nitrate contamination on the analysis of carbon and oxygen isotopes in carbonate and a low-temperature phosphoric acid digestion approach for online sample preparation.

High-temperature phosphoric acid digestion combined with continuous-flow isotope ratio mass spectrometry (CF-IRMS) is one of the standard methods for the determination of carbon and oxygen isotopes in carbonate. However, the routine purification protocol has not been tested by samples with nitrate contamination. Here we show that the standard protocol is not capable of measuring such samples with satisfactory accuracy and precision. We tested the low-temperature (25°C) phosphoric acid digestion method combined with CF-IRMS for the measurement of natural carbonate samples with ~1% nitrate concentration and nitrate-doped carbonate standards. We also recorded the Raman spectra of the gases extracted from the nitrate-doped carbonate standards to identify the gas species that affect the results. The accuracy and precision of δ18 O values for nitrate-doped carbonate standards digested at 72°C are much poorer than routine measurements. For the measurements of doped standards reacted at 25°C, the accuracy and precision of the δ13 C and δ18 O values are indistinguishable from normal results. Raman spectra demonstrate that NO2 generated from high-temperature digestion is the possible interference species for oxygen isotope measurement. Low-temperature digestion is a reliable method for the measurement of carbon and oxygen isotopes in nitrate-contaminated carbonate samples because the formation of NO2 can be largely reduced. This approach can be applied to some nitrate-rich speleothems, evaporites and other samples with nitrate contamination.

'Lake Gorgana' - A paleolake in the Lower Danube Valley revealed using multi-proxy and regionalisation approaches

Geoarchaeological research at the archaeological site of Mǎgura Gorgana should elucidate the environmental setting Neolithic and Copper Age communities were faced with, when they settled along the Lower Danube Valley, Romania. This should enable a better understanding of human-environment interactions, the natural preconditions of a trading network, as well as the living conditions of the human population during this time. To reach this aim numerous corings were conducted in the study area and the sediments were analysed using a multitude of methods, as total element analyses, determination of carbon and nitrogen isotopes, radiocarbon and OSL-dating and microfaunistical analyses. The present study shows how the interpretation of the results applying a multi-proxy and a ‘regionalisation’ approach reveals the existence and the extent of ‘Lake Gorgana’, a vast paleolake that covered nearly the whole floodplain in the study area during a long phase of the Holocene and, in particular, during the settlement period at Mǎgura Gorgana in the 5th millennium BC. These new findings are of great importance for archaeological concerns but the lake sediments even more represent a valuable geoarchive containing information about e.g. changing lake ecology, paleoclimate, and the human footprint with regard to different spatial and temporal scales.

Determination of carbon isotope ratios for honey samples by means of a liquid chromatography/isotope ratio mass spectrometry system coupled with a post-column pump.

Liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) has been used to authenticate and trace products such as honey, wine, and lemon juice, and compounds such as caffeine and pesticides. However, LC/IRMS has several disadvantages, including the high cost of the CO2 membrane and blocking by solidified sodium persulfate. Here, we developed an improved system for determining carbon isotope ratios using LC/IRMS. The main improvement was the use of a post-column pump. Using the improved system, we determined δ13 C values for glucose with high accuracy and precision (0.1‰ and 0.1‰, respectively; n=3). The glucose, fructose, disaccharide, trisaccharide, and organic acid constituents of honey samples were analyzed using LC/IRMS. The δ13 C values for glucose, fructose, disaccharides, trisaccharides, and organic acids ranged from -27.0 to -24.2‰, -26.8 to -24.0‰, -28.8 to -24.0‰, -27.8 to -22.8‰, and-30.6 to -27.4‰, respectively. The analysis time was a third to a half of that required for analysis by previously reported methods. The column flow rate could be arbitrarily adjusted with the post-column pump. We applied the improved method to 26 commercial honey samples. Our results can be expected to be useful for other researchers who use LC/IRMS.

In Situ Carbon Isotope Analysis by Laser Ablation MC-ICP-MS.

Carbon isotopes have been widely used in tracing a wide variety of geological and environmental processes. The carbon isotope composition of bulk rocks and minerals was conventionally analyzed by isotope ratio mass spectrometry (IRMS), and, more recently, secondary ionization mass spectrometry (SIMS) has been widely used to determine carbon isotope composition of carbon-bearing solid materials with good spatial resolution. Here, we present a new method that couples a RESOlution S155 193 nm laser ablation system with a Nu Plasma II MC-ICP-MS, with the aim of measuring carbon isotopes in situ in carbonate minerals (i.e., calcite and aragonite). Under routine operating conditions for δ13C analysis, instrumental bias generally drifts by 0.8‰-2.0‰ in a typical analytical session of 2-3 h. Using a magmatic calcite as the standard, the carbon isotopic composition was determined for a suite of calcite samples with δ13C values in the range of -6.94‰ to 1.48‰. The obtained δ13C data are comparable to IRMS values. The combined standard uncertainty for magmatic calcite is <0.3‰ (1s). No significant matrix effects have been identified in calcite with the amplitude of chemical composition variation (i.e., MnO, SrO, MgO, or FeO) up to 2.5 wt %. Two modern corals were investigated using magmatic calcite as the calibration standard, and the average δ13C values for both corals are similar to the bulk IRMS values. Moreover, coral exhibits significant heterogeneity in carbon isotope compositions, with differences up to 4.85‰ within an individual coral. This study indicates that LA-MC-ICP-MS can serve as an appropriate method to analyze carbon isotopes of carbonate minerals in situ.

Evolution of chemical and isotopic composition of inorganic carbon in a complex semi-arid zone environment: Consequences for groundwater dating using radiocarbon

Estimating groundwater age is important for any groundwater resource assessment and radiocarbon (14C) dating of dissolved inorganic carbon (DIC) can provide this information. In semi-arid zone (i.e. water-limited environments), there are a multitude of reasons why 14C dating of groundwater and traditional correction models may not be directly transferable. Some include; (1) the complex hydrological responses of these systems that lead to a mixture of different ages in the aquifer(s), (2) the varied sources, origins and ages of organic matter in the unsaturated zone and (3) high evaporation rates. These all influence the evolution of DIC and are not easily accounted for in traditional correction models. In this study, we determined carbon isotope data for; DIC in water, carbonate minerals in the sediments, sediment organic matter, soil gas CO2 from the unsaturated zone, and vegetation samples. The samples were collected after an extended drought, and again after a flood event, to capture the evolution of DIC after varying hydrological regimes. A graphical method (Han et al., 2012) was applied for interpretation of the carbon geochemical and isotopic data. Simple forward mass-balance modelling was carried out on key geochemical processes involving carbon and agreed well with observed data. High values of DIC and δ13CDIC, and low 14CDIC could not be explained by a simple carbonate mineral–CO2 gas dissolution process. Instead it is suggested that during extended drought, water–sediment interaction leads to ion exchange processes within the top ∼10–20m of the aquifer which promotes greater calcite dissolution in saline groundwater. This process was found to contribute more than half of the DIC, which is from a mostly ‘dead’ carbon source. DIC is also influenced by carbon exchange between DIC in water and carbonate minerals found in the top 2m of the unsaturated zone. This process occurs because of repeated dissolution/precipitation of carbonate that is dependent on the water salinity driven by drought and periodic flooding conditions. This study shows that although 14C cannot be directly applied as a dating tool in some circumstances, carbon geochemical/isotopic data can be useful in hydrological investigations related to identifying groundwater sources, mixing relations, recharge processes, geochemical evolution, and interaction with surface water.

Distribution of iridium and associated geochemistry across the Triassic–Jurassic boundary in sections at Kuhjoch and Kendlbach, Northern Calcareous Alps, Austria

Samples from strata spanning the Triassic–Jurassic boundary at the GSSP at Kuhjoch and at Kendlbachgraben were studied by NAA, XRF and combustion analysis to determine Ir levels and associated geochemistry. The results are compared to previously determined carbon isotope stratigraphy at these sections. Ir concentrations in the limestones of the Kössen Formation at Kuhjoch are very low (<10pg/g) below the top of the formation, rising to 26pg/g, in the T-bed at the top of the Eiberg Member. The Tiefengraben Member of the Kendlbach Formation exhibits higher concentrations of Ir in general relative to the strata below. The shift to higher levels is abrupt at the base of the member, coinciding with a decrease in carbonate content. Concentrations of 60 to 80pg/g are typical through the entire thickness of the Schattwald Beds and into the gray Tiefengraben Member strata, peaking at 145pg/g. Concentrations decline to 30–40pg/g above 680cm from the formation base, coinciding with increasing carbonate content. The analyses from the Kendlbachgraben section compare well with those from Kuhjoch, with similar difference in Ir concentration between the Kössen and Kendlbach formations. In both sections, the initial increase in Ir corresponds to the initial carbon isotope excursion. Concentrations of redox-sensitive elements indicate transient reducing conditions during deposition of the uppermost Kössen Formation, but oxidizing conditions during Tiefengraben Member deposition. The Al/Ti ratio indicates more intense weathering during deposition of the lowermost 20cm of the Tiefengraben Member, but otherwise consistently humid to sub-humid climate prevailed during deposition. The primary control on Ir concentration in the sampled section is formation lithology, although there are variations within the Tiefengraben Member that are independent of carbonate content. Enrichment of Ir at the top of the T-bed is associated with a redox boundary, but the cause of other variations is undetermined.

Root‐related rhodochrosite and concretionary siderite formation in oxygen‐deficient conditions induced by a ground‐water table rise

AbstractSedimentological, mineralogical, stable carbon and oxygen isotope determinations and biomarker analyses were performed on siderite concretions occurring in terrestrial silts to understand their formation and to characterize the sedimentary and diagenetic conditions favouring their growth. High δ13C values (6·4‰ on average) indicate that siderite precipitated in an anoxic environment where bacterial methanogenesis operated. The development of anoxic conditions during shallow burial was induced by a change in sedimentary environment from flood plain to swamp, related to a rise of the ground‐water table. Large amounts of decaying plant debris led to efficient oxygen consumption within the pore‐water in the peat. Oxygen depletion, in combination with a decrease in sedimentation rate, promoted anoxic diagenetic conditions under the swamp and favoured abundant siderite precipitation. This shows how a change in sedimentary conditions can have a profound impact on the early‐diagenetic environment and carbonate authigenesis. The concretions contain numerous rhizoliths; they are cemented with calcium‐rhodochrosite, a feature which has not been reported before. The rhodochrosite cement has negative δ13C values (−16·5‰ on average) and precipitated in suboxic conditions due to microbial degradation of roots coupled to manganese reduction. The exceptional preservation of the epidermis/exodermis and xylem vessels of former root tissues indicates that the rhodochrosite formed shortly after the death of a root in water‐logged sediments. Rhodochrosite precipitated during the initial stages of concretionary growth in suboxic microenvironments within roots, while siderite cementation occurred simultaneously around them in anoxic conditions. These suboxic microenvironments developed because oxygen was transported from the overlying oxygenated soil into sediments saturated with anoxic water via roots acting as permeable conduits. This model explains how separate generations of carbonate cements having different mineralogy and isotopic compositions, which would conventionally be regarded as cements precipitated sequentially in different diagenetic zones during gradual burial, can form simultaneously in shallow burial settings where strong redox gradients exist around vertically oriented permeable root structures.

Laser ablation molecular isotopic spectrometry of carbon isotopes

Quantitative determination of carbon isotopes using Laser Ablation Molecular Isotopic Spectrometry (LAMIS) is described. Optical emission of diatomic molecules CN and C2 is used in these measurements. Two quantification approaches are presented: empirical calibration of spectra using a set of reference standards and numerical fitting of a simulated spectrum to the experimental one. Formation mechanisms of C2 and CN in laser ablation plasma are briefly reviewed to provide insights for implementation of LAMIS measurements. A simulated spectrum of the 12C2 Swan system was synthesized using four constituents within 473.5–476.5nm. Simulation included three branches of 12C2 (1-0), branches R(0-0) and R(1-1), and branch P(9-8) of 12C2. Spectral positions of the tail lines in R(0-0) and R(1-1) were experimentally measured, since they were not accurately known before. The Swan band (1-0) of the isotopologue 13C12C was also simulated. Fitting to the experimental spectrum yielded the ratio 13C/12C=1.08% in a good agreement with measurements by isotope ratio mass spectrometry. LAMIS promises to be useful in coal, oil and shale exploration, carbon sequestration monitoring, and agronomy studies.

GC/MS method for determining carbon isotope enrichment and concentration of underivatized short-chain fatty acids by direct aqueous solution injection of biogas digester samples

In anaerobic digestion of organic matter, several metabolic pathways are involved during the simultaneous production and consumption of short-chain fatty acids (SCFA) in general and acetate in particular. Understanding the role of each pathway requires both the determination of the concentration and isotope enrichment of intermediates in conjunction with isotope labeled substrates. The objective of this study was to establish a rapid and simple GC/MS method for determining the isotope enrichment of acetate and concentration of underivatized short-chain fatty acids (SCFA) in biogas digester samples by direct liquid injection of acidified aqueous samples. Sample preparation involves only acidification, centrifugation and filtration of the aqueous solution followed by direct injection of the aqueous supernatant solution onto a polar column. With the sample preparation and GC/MS conditions employed, well-resolved and sharp peaks of underivatized SCFA were obtained in a reasonably short time. Good recovery (96.6–102.3%) as well as low detection (4–7µmol/L) and quantification limits (14–22µmol/L) were obtained for all the 6 SCFA studied. Good linearity was achieved for both concentration and isotope enrichment measurement with regression coefficients higher than 0.9978 and 0.9996, respectively. The method has a good intra- and inter-day precision with a relative standard deviation (RSD) below 6% for determining the tracer-to-tracee ratio (TTR) of both [2-13C]acetate and [U-13C]acetate. It has also a good intra– and inter-day precision with a RSD below 6% and 5% for determining the concentration of standard solution and biogas digester samples, respectively. Acidification of biogas digester samples with oxalic acid provided the low pH required for the protonation of SCFA and thus, allows the extraction of SCFA from the complex sample matrix. Moreover, oxalic acid was the source of formic acid which was produced in the injector set at high temperature. The produced formic acid prevented the adsorption of SCFA in the column, thereby eliminating peak tailing and ghost peaks. The applicability of the validated GC/MS method for determining the concentration of acetate and its 13C isotope enrichment in anaerobic digester samples was tested and the results demonstrated the suitability of this method for identifying the metabolic pathways involved in degradation and production of acetate.

Phylogeny of Sesuvioideae (Aizoaceae) – Biogeography, leaf anatomy and the evolution of C4 photosynthesis

Sesuvioideae (Aizoaceae) form a small subfamily of drought-tolerant plants exhibiting leaf succulence, halophytic ecology, and the C 4 photosynthetic pathway. Sesuvioideae are sister to the species-rich subfamilies Ruschioideae, Mesembryanthemoideae and Aizooideae that contain many CAM lineages. This close relationship of CAM and C 4 taxa identifies the Sesuvioideae as an important clade to address hypotheses of photosynthetic pathway evolution. This study presents a molecular phylogeny of Sesuvioideae based on five markers ( atpB-rbcL spacer, rps16 intron, trnL - trnF spacer, petB - petD spacer, ITS) and 51 accessions representing all genera and 37 species. We determined carbon isotope data of 103 samples and examined the leaf anatomy of 25 species. A RASP (Reconstruct Ancestral State in Phylogenies) analysis was used to reconstruct the ancestral biogeography of Sesuvioideae and trace its current worldwide distribution. Maximum likelihood character optimization was conducted for six traits related to photosynthetic type and leaf anatomy to characterize the evolution of leaf types in Sesuvioideae. The well-resolved molecular phylogeny revealed an African/Arabian origin of the subfamily with Tribulocarpus as sister to a clade containing Trianthema , Sesuvium , Cypselea and Zaleya . Intercontinental dispersal occurred within Trianthema (to Australia and South America) and Sesuvium / Zaleya (to Australia and North/Central America). Character optimizations favoured a single origin of C 4 photosynthesis with two reversions to the C 3 state, one within American Sesuvium and the other in Trianthema subgenus Trianthema . However, biochemical diversity of the C 4 syndrome in Sesuvioideae might indicate multiple origins of the C 4 pathway. Two C 3 and four C 4 anatomical types (atriplicoid, salsoloid, portulacelloid and pilosoid) are present in the subfamily, based on differences in water storage tissue, vascular bundle arrangements, and chlorenchyma structure. Ancestral state reconstruction indicates multiple losses or reduction of water storage tissue in the subfamily and frequent shifts in leaf anatomical traits.

Aquatic Resources in Foodcrusts: Identification and Implication

Foodcrusts, the charred surface deposits on pottery vessel surfaces, provide a rich source of data regarding container function. This article reviews recent applications focusing on the detection of aquatic resources (marine and freshwater) in pottery vessels using a range of analytical approaches including bulk isotope measurements of carbon and nitrogen, lipid biomarker analysis, and compound-specific carbon isotope determinations. Such data can help to evaluate the presence of reservoir effects when undertaking radiocarbon dating of foodcrust samples. In particular, molecular and isotopic analysis can aid in the selection of suitable candidates for14C where it can be demonstrated that aquatic resources are unlikely to contribute to the residue. Prospects for compound-specific14C analysis of lipids in foodcrusts and ceramic-absorbed residues are also discussed.

Evaluating degradation of hexachlorcyclohexane (HCH) isomers within a contaminated aquifer using compound-specific stable carbon isotope analysis (CSIA)

The applicability of compound-specific stable carbon isotope analysis (CSIA) for assessing biodegradation of hexachlorocyclohexane (HCH) isomers was investigated in a contaminated aquifer at a former pesticide processing facility. A CSIA method was developed and tested for efficacy in determining carbon isotope ratios of HCH isomers in groundwater samples using gas chromatography – isotope ratio mass spectrometry (GC-IRMS). The carbon isotope ratios of HCHs measured for samples taken from the field site confirmed contaminant source zones at former processing facilities, a storage depot and a waste dump site. The 13C-enrichment in HCHs provided evidence for biodegradation of HCHs especially downstream of the contaminant source zones. CSIA from monitoring campaigns in 2008, 2009 and 2010 revealed temporal trends in HCH biodegradation. Thus, the impact and progress of natural attenuation processes could be evaluated within the investigated aquifer. Calculations based on the Rayleigh-equation approach yielded levels of HCH biodegradation ranging from 30 to 86 %. Moreover, time- and distance-dependent in situ first-order biodegradation rate constants were estimated with maximal values of 3 × 10−3 d−1 and 10 × 10−3 m−1 for α-HCH, 11 × 10−3 d−1 and 37 × 10−3 m−1 for β-HCH, and 6 × 10−3 d−1 and 19 × 10−3 m−1 for δ-HCH, respectively. This study highlights the applicability of CSIA for the assessment of HCH biodegradation within contaminated aquifers.