Carbon Isotope Fractionation Factors Research Articles

Geochemical evidence for biodegradation in high-rank coals from Qinshui Basin, North China

Understanding the generation of secondary microbial methane (SMM) is important for the evaluation of natural gas resources and instructive for the stimulation of methane production. Coal seams are popular targets for extracting in situ preserved methane and studying microbe-stimulated methane yield. However, few studies have been done on overmature coals. Here we collected gas samples from coals varying from bituminous to meta-anthracite in the Qinshui Basin, North China, and analysed the molecular and stable isotopic compositions to systematically evaluate the influence of biodegradation in high-rank coals in geological settings. The stable isotope signatures (δ13C and δD) of methane are dominated by the thermal decomposition of organic matter in deep coals but inconsistent with the maturity rank of shallow burial coals. The decoupling of coal maturity with C1/C2+ ratios and δ13C-CH4 values, and positive δ13C-CO2 values (−9.2 to +24.4 ‰) suggest biodegradation of light wet gases (C2+ components) and CO2 reduction. Negative trends between δ13C-CO2 and CH4/CO2 in shallow burial coal seams reveal the mutual conversion of CH4 and CO2 and carbon isotope exchange, driven by microorganisms. The calculated isotopic temperatures (33–328 °C) based on the carbon isotope fractionation factors between CH4 and CO2 (1.024–1.069) demonstrate that carbon isotope exchange is prevalent in high-rank coals. It also reveals that the burial depth is an imperative factor in controlling microbial environments and thus the biodegradation process. This study implicates the potential of high-rank coals as the target for microbial-enhanced methane recovery and also implies that microorganisms are widely involved in reservoir carbon cycling.

Is ebullition or diffusion more important as methane emission pathway in a shallow subsaline lake?

Methane (CH4) emissions via ebullition contribute significantly to greenhouse gas emissions from freshwater bodies. According to the literature, the ebullition pathway may even be the most important pathway in some cases, particularly in shallow lakes. Ebullition rates are not often estimated because of the high uncertainty associated with episodic releases, leading to difficulties in their determination. This study provides an estimate of such emissions in a large, shallow, subsaline lake in eastern Austria, Lake Neusiedl, and compares them to the diffusion pathway. Ebullition gas sampling was conducted every 5–10 days over a period of 107 days from late March to mid-July 2021, using ebullition traps placed in three distinct locations: Reed belt, Channel and Open water/Lake. The aim was to study the temporal and spatial heterogeneity of ebullition and its contribution to total emissions. At the same time, several water quality and other environmental parameters were measured and then tested against the CH4 ebullition rates to explore them as potential drivers for this pathway. The carbon isotope fractionation factor (αC) of the measured CH4 ebullition gas, ranging from 1.03 to 1.06, indicates a dominance of the acetoclastic methanogenesis in the sediments of Lake Neusiedl, regardless of the location. The Reed belt location showed the highest mean CH4 ebullition rate (17 ± 28 mg CH4 m−2 d−1), which is >340-fold higher than the mean of the other two locations, and demonstrated also a strong temperature dependency. In all locations at Lake Neusiedl, the median CH4 fluxes via diffusion are significantly higher than via ebullition. Our analyses do not confirm the dominance of the ebullition pathway in any of the studied locations. Whereas at the Reed belt, ebullition accounts for 48 % of the CH4 emissions, in the other two locations, is responsible only for about 1 %.

Carbon and hydrogen isotope fractionation of phthalates during photocatalysis reactions in aqueous solution containing Fe(III) complexes or iron minerals

The hydrogen and carbon isotope fractionation factor (ε2H, ε13C) of dimethyl-, diethyl‑ and dibutyl phthalic acid ester during photosensitized degradation by artificial sunlight with Fe(III) ions and iron minerals (hematite, goethite and magnetite) in aqueous solution were examined by compound-specific isotope analysis (CSIA) in order to analyze the degradation mechanism. Hematite does not catalyze photosensitized degradation of phthalates. The correlation of 2H and 13C isotope fractionation (Λ = Δδ2H/Δδ13C) of phthalates with increasing chain length (dimethyl-; diethyl‑; and dibutyl-) were compared with values of the ∙OH radical model reaction with the aromatic ring as well as acidic and alkaline hydrolysis. The Λ values of die photosensitized reaction of diethyl phthalate with goethite (-5.1 ± 1.8) and magnetite (-18.9 ± 3.9) show a large difference compared to Fe(III) solutions (4.7 ± 0.9 to 4.8 ± 1.0) suggesting specific reaction mechanisms. The fractionation factors determined here have potential to characterize the degradation of phthalates catalyzed by photo-induced reaction of Fe(III), goethite and magnetite in natural system and in remediation approaches.

Carbon isotope effects in the sorption of chlorinated ethenes on biochar and activated carbon

As an alternative to activated carbon, biochar is a promising, environmentally friendly sorbent that can be used to remove organic groundwater pollutants, such as chlorinated ethenes (CEs). Stable isotope fractionation in biofilters is used to quantify pollutant degradation and to distinguish degradation from pollutant sorption on e.g. biochar. However, the sorption of CEs on biochar, and the potential abiotic fractionation processes remain to be tested. The sorption process of CEs and ethene on activated carbon and biochar was investigated with regard to the isotope effects for the differentiation from microbial degradation processes. Results from physical and chemical characterization of biochar indicated that biochar feedstock and pyrolysis conditions determined sorption performance depending on the surface chemistry and the pore size distribution of the coarse sorbent particles. The sorption capacity of the activated carbon was significantly higher with highly chlorinated ethenes, but similar to the biochars with low chlorination. Apparent carbon isotope fractionation factors (ε) of +0.1 to −4.4 ‰ were found above measurement uncertainties of GC/IRMS. The extent of isotope enrichment of the 13C bearing isotopologues in the residual aqueous phase (ε < 0) was characteristic for individual pairs of pollutant and sorbent material and could be related to pore-filling processes limited by the micropore size distribution of sorbent materials and the chemical properties of sorbed pollutants. Especially the large isotope fractionation during the sorption of ethene led to the assumption that diffusion processes within the pore matrix of the sorbent particles contributed to the observed isotope effects, but should still be considered a property of sorption. Concluding on the results indicated that sorption processes can have a significant contribution to carbon isotope fractionation in CEs and ethene. These should not be neglected in the evaluation of biofilters for groundwater purification, in which CEs are simultaneously degraded by microbes.

Carbon and hydrogen stable isotope fractionation due to monooxygenation of short-chain alkanes by butane monooxygenase of Thauera butanivorans Bu-B1211.

Multi element compound-specific stable isotope analysis (ME-CSIA) is a tool to assess (bio)chemical reactions of molecules in the environment based on their isotopic fingerprints. To that effect, ME-CSIA concepts are initially developed with laboratory model experiments to determine the isotope fractionation factors specific for distinct (bio)chemical reactions. Here, we determined for the first time the carbon and hydrogen isotope fractionation factors for the monooxygenation of the short-chain alkanes ethane, propane, and butane. As model organism we used Thauera butanivorans strain Bu-B1211 which employs a non-haem iron monooxygenase (butane monooxygenase) to activate alkanes. Monooxygenation of alkanes was associated with strong carbon and hydrogen isotope effects: εbulkC = -2.95 ± 0.5 ‰ for ethane, -2.68 ± 0.1 ‰ for propane, -1.19 ± 0.18 ‰ for butane; εbulkH = -56.3 ± 15 ‰ for ethane, -40.5 ± 2.3 ‰ for propane, -14.6 ± 3.6 ‰ for butane. This resulted in lambda (Λ ≈ εHbulk/εCbulk) values of 16.2 ± 3.7 for ethane, 13.2 ± 0.7 for propane, and 11.4 ± 2.8 for butane. The results show that ME-CSIA can be used to track the occurrence and impact of monooxygenase-dependent aerobic processes converting short-chain alkanes in natural settings like marine and terrestrial seeps, gas reservoirs, and other geological formations impacted by natural gas.

Carbon isotope fractionation of di-(2-ethylhexyl)-phthalate during photosensitized degradation by •OH and [formula omitted] for characterization of reaction mechanisms

Compound specific isotope analysis (CSIA) has been used to explore the degradation pathways of di-(2-ethylhexyl)-phthalate (DEHP) in photosensitization experiments with H2O2, TiO2, Fe(III) and PMS. The reaction kinetics could be described by first order rate constants [k (h−1)]. The highest degradation rates of DEHP were observed in UV/TiO2 catalyzed reactions.Carbon isotope fractionation factors (εC) and metabolite patterns were examined to analyze the radical reaction. Similar carbon isotope fractionation factors have been found in experiments dominated by OH radical reactions (UV/H2O2, −1.2 ± 0.3‰; UV/TiO2, −1.2 ± 0.5‰). An inverse carbon isotope fractionation has been found for UV/Fe(III) (0.8 ± 0.2‰) and SO4− (0.6 ± 0.2‰ at pH 3) catalyzed reactions indicating that these reactions preferentially taking place at the H atom bond at tert-butyl carbon of the side chain. This hypothesis is further supported by an experiment with isobutyl acetate (εC = 1.8 ± 0.2‰ for UV/Fe(III) at pH 7; εC = -1.1 ± 0.2‰ for UV/PMS at pH 6 and εC = 0.9 ± 0.3‰ for UV/PMS at pH 3).Quenching experiments in the UV/PMS catalyzed reaction indicated a coexistence of SO4− and OH at pH 7. SO4− was the predominant radical at pH 3. OH was found to be the predominant radical in UV/H2O2 and UV/TiO2 reactions. CSIA indicated that the addition of OH to the aromatic ring of DEHP by the RAF pathway is the main reaction mechanism in UV/H2O2 and UV/TiO2 experiments. The SO4− attacks preferentially the C-H bond at the tert-butyl carbon position of the side chain of DEHP which causes an inverse carbon isotope effect. Thus, carbon isotope enrichment factors can be used to distinguish different radical reaction mechanisms with DEHP as a substrate.

Temperature and organic carbon quality control the anaerobic carbon mineralization in peat profiles via modulating microbes: A case study of Changbai Mountain

Peatlands account for a significant fraction of the global carbon stock. However, the complex interplay of abiotic and biotic factors governing anaerobic carbon mineralization in response to warming remains unclear. In this study, peat sediments were collected from a typical northern peatland-Changbai Mountain to investigate the behavior and mechanism of anaerobic carbon mineralization in response to depth (0–200 cm) and temperature (5 °C, 15 °C and 20 °C), by integrating geochemical and microbial analysis. Several indices including humification indexes (HI), aromaticity, and water extractable organic carbon (WEOC) components were applied to evaluate carbon quality, while 16S rRNA sequencing was used to measure microbial composition. Regardless of temperature, degradations of carbon quality and associated reduction in microbial abundance as well as diversity resulted in a decrease in anaerobic carbon mineralization (both CO2 and CH4) towards greater depth. Warming either from 5 °C to 15 °C or 20 °C significantly increased anaerobic carbon mineralization in all depth profiles by improving carbon availability. Enhanced carbon availabilities were mediated by the change in microbial composition (p < 0.01) and an increase in metabolic activities, which was particularly evident in the enhanced β-glucosidase activity and microbial collaborations. A remarkable increase of over 10-fold in the relative abundance of the Geothrix genus was observed under warming. Overall, warming resulted in an enhanced contribution of CH4 emission and a higher ratio of hydrogenotrophic methanogenesis, as evidenced by carbon isotope fractionation factors. In addition, deep peat soils (>100 cm) with recalcitrant carbon demonstrated greater temperature sensitivity (Q10: ∼2.0) than shallow peat soils (Q10:∼1.2) when temperature increased from 15 °C to 20 °C. The findings of this study have significantly deepened our understanding for mechanisms of carbon quality and microbe-driven anaerobic carbon mineralization in peatlands under global warming.

Bacterial physiology highlighted by the δ13C fractionation of bacteriohopanetetrol isomers

Lipid biomarkers, such as the various bacteriohopanetetrol (BHT) isomers studied here, are useful tools in tracing bacterially mediated nitrogen and carbon cycle processes affecting greenhouse gas emissions, including the anaerobic oxidation of ammonia. Three BHT isomers occur commonly in the environment. By gas chromatography, BHT-34S elutes first; it is produced by numerous bacteria. The two later eluting isomers are more constrained in their origin. The marine anammox bacteria ‘Ca. Scalindua’ is the only known producer of a BHT isomer of unknown stereochemistry (BHT-x), making BHT-x a diagnostic biomarker in anoxic marine settings. The BHT-34R isomer is produced by three freshwater aerobic heterotrophic producers (Frankia spp., Acetobacter pasteurianus, and Komagataeibacter xylinus), a freshwater serine-cycle (Type II) methanotroph (Methylocella palustris), and the freshwater anammox ‘Ca. Brocadia’, which makes the detection of freshwater anammox using BHT-34R more complicated. We investigated whether the source of BHT-34R in freshwater environments could be ascertained via its δ13C value. We used conventional on-column gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) (as opposed to high temperature GC-C-IRMS) to determine the δ13C composition of acetylated BHT isomers in cultured bacteria and bacterial enrichments. We combined these with bulk biomass and substrate δ13C compositions to establish carbon isotopic fractionation factors. The two anammox genera had large fractionation factors from dissolved inorganic carbon (DIC) to biomass (Δ13Cbiomass – DIC = –43.8 to –26.4 ‰) and to BHTs (Δ13CBHT – DIC = –53.8 to –38.2 ‰), which clearly distinguished them from the freshwater aerobic heterotrophic producers (Δ13Cbiomass – substrate = –2.3 to –0.1 ‰; Δ13CBHT – substrate = –12.8 to 5.2 ‰). Methylocella assimilated mainly carbon from DIC, rather than from methane, into its biomass and BHT, and previous work suggested this assimilation comes with relatively small fractionation. Thus, in peatlands, the BHT δ13C values of Methylocella would not reflect the low δ13C values of biogenic methane. Consequently, the presence of BHT-34R with low δ13C values relates to ‘Ca. Brocadia’ and presents a novel tool to trace anammox in freshwater environments.

Sulfate concentrations affect sulfate reduction pathways and methane consumption in coastal wetlands

Coastal wetlands are an important source of methane emissions, and understanding the mechanisms that control methane emissions from coastal wetlands is of great significance to global warming. Anaerobic oxidation of methane driven by sulfate is an important process to prevent methane emissions from coastal wetlands. The effects of environmental changes on this process and the function of the sulfate-methane transition zone (SMTZ) are poorly understood. In this study, spatiotemporal variations in pore-water geochemistry (concentrations of SO42-, CH4 and DIC as well as δ13C-DIC and δ13C-CH4) in the Beidagang wetland, Tianjin, China, were investigated to unravel factors controlling the role of anaerobic oxidation of methane in coastal wetlands. Results show that the geochemical profile of pore-water is characterized by significant spatial and temporal variability, which may be related to changes in sulfate concentration, temperature and dissolved oxygen. The carbon isotope fractionation factors (εC) during methane oxidation range from 8.9‰ to 12.5‰, indicating that the sulfate-driven anaerobic oxidation of methane (S-AOM) dominates the methane oxidation in the Beidagang coastal wetland in both winter and summer, in both high and low salinity wetlands, and in both open water and littoral areas. However, sulfate concentration has a strong influence on the sulfate reduction pathways and methane consumption. The consumption of methane and sulfate by S-AOM is more significant in coastal wetlands with high sulfate concentrations, with S-AOM consuming nearly all of the upward-diffusing methane (96%) and downward-diffusing sulfate (96%). In addition, the dissolved inorganic carbon (DIC) produced in the pore-water mainly comes from methanogenesis, accounting for more than 80% of the total DIC pool, but in the areas with high sulfate concentrations in water column, the contribution of S-AOM to the DIC pool is greater, although only a small fraction of the total DIC pool (9%). The depth and width of the SMTZ show a clear spatial and temporal pattern, with active methanogenesis activity and upward high methane flux shoaling the SMTZ and increasing the risk of high methane emissions from coastal wetlands with low sulfate concentrations. Our findings highlight the importance of sulfate-driven anaerobic oxidation of methane in coastal wetlands and the effect of sulfate concentration on it. It contributes to our understanding of the mechanism of methane production and emissions from the coastal wetland system, particularly in light of the increased demand for coastal wetland restoration under global warming.

Assessment of aerobic biodegradation of lower-chlorinated benzenes in contaminated groundwater using field-derived microcosms and compound-specific carbon isotope fractionation

Biodegradation of lower chlorinated benzenes (tri-, di- and monochlorobenzene) was assessed at a coastal aquifer contaminated with multiple chlorinated aromatic hydrocarbons. Field-derived microcosms, established with groundwater from the source zone and amended with a mixture of lower chlorinated benzenes, evidenced biodegradation of monochlorobenzene (MCB) and 1,4-dichlorobenzene (1,4-DCB) in aerobic microcosms, whereas the addition of lactate in anaerobic microcosms did not enhance anaerobic reductive dechlorination. Aerobic microcosms established with groundwater from the plume consumed several doses of MCB and concomitantly degraded the three isomers of dichlorobenzene with no observable inhibitory effect. In the light of these results, we assessed the applicability of compound stable isotope analysis to monitor a potential aerobic remediation treatment of MCB and 1,4-DCB in this site. The carbon isotopic fractionation factors (ε) obtained from field-derived microcosms were -0.7‰ ± 0.1 ‰ and -1.0‰ ± 0.2 ‰ for MCB and 1,4-DCB, respectively. For 1,4-DCB, the carbon isotope fractionation during aerobic biodegradation was reported for the first time. The weak carbon isotope fractionation values for the aerobic pathway would only allow tracing of in situ degradation in aquifer parts with high extent of biodegradation. However, based on the carbon isotope effects measured in this and previous studies, relatively high carbon isotope shifts (i.e., ∆δ13C > 4.0 ‰) of MCB or 1,4-DCB in contaminated groundwater would suggest that their biodegradation is controlled by anaerobic reductive dechlorination.

Simultaneous Calculation of Chemical and Isotope Equilibria Using the GEOCHEQ_Isotope Software: Oxygen Isotopes

—The GEOCHEQ_Isotope software package, elaborated previously for modeling chemical and carbon isotope equilibria in hydrothermal and hydrogeochemical systems by minimizing the Gibbs energy, is extended to the simultaneous calculation of carbon and oxygen isotopic effects. Similar to what was done for carbon, the β-factor formalism was used to develop algorithms and a database for calculating the isotopic effects of oxygen. According to the developed algorithm, the Gibbs energy of formation of a rare isotopologue, G*(P, T), is calculated through the Gibbs energy of formation of the main isotopologue, the value of the β18O factor of this substance, and the mass ratio of the rare (18O) and main (16O) isotopes. The isotope mixture is assumed to be ideal. The temperature dependence of the β-factor is unified as a polynomial in reciprocal absolute temperature. Necessary information on oxygen isotope equilibria involving important geochemical compounds was critically analyzed, and the available data were reconciled and modified. The temperature dependences of the β18O-factors were correspondingly optimized. The thermodynamic database was updated by adding information on the temperature dependence of β18O-factors specified by polynomial coefficients for each substance. The usage of the GEOCHEQ_Isotope software package and the corresponding database is demonstrated by modeling the dependence of oxygen and carbon isotope fractionation factors on the acidity of the solution (pH) in a carbonate hydrothermal system. The simulation results are in a good agreement with experimental data available from the literature. The enrichment of dissolved carbonates in the 18O heavy oxygen isotope relative to water decreases with increasing pH of the system. At the same time, a pH increase results in a decrease in the negative carbon isotope shift between calcite and dissolved carbonates. At high pH values (~11), the isotope shift inversion and the enrichment of the dissolved carbonate in the heavy carbon isotope relative to calcite are predicted.

Hydrogen and carbon isotope fractionation factors of aerobic methane oxidation in deep-sea water

Abstract. Isotope fractionation factors associated with various biogeochemical processes are important in ensuring the reliable use of isotope tracers in biogeosciences at large. Methane is a key component of the subsurface biosphere and a notable greenhouse gas, making the accurate evaluation of methane cycles, including microbial methanotrophy, imperative. Although the isotope fractionation factors associated with methanotrophy have been examined under various conditions, the dual-isotope fractionation factors of aerobic methanotrophy in oxic seawater remain unclear. Here, we investigated hydrogen and carbon isotope ratios of methane as well as the relevant biogeochemical parameters and microbial community compositions in hydrothermal plumes in the Okinawa Trough. Methanotrophs were found to be abundant in plumes above the Hatoma Knoll vent site, and we succeeded in simultaneously determining hydrogen and carbon isotope fractionation factors associated with the aerobic oxidation of methane (εH=49.4±5.0 ‰, εC=5.2±0.4 ‰) – the former being the first of its kind ever reported. This εH value is comparable with values reported from terrestrial ecosystems but clearly lower than those from aerobic and anaerobic methanotroph enrichment cultures, as well as incubations of methanotrophic isolates. The covariation factor between δ13CCH4 and δDCH4, Λ (9.4 or 8.8 determined using two different methods), was consistent with those from methanotrophic isolate incubations. These values are valuable for understanding dynamics of methane cycling in the marine realm, and future applications of the approach to other habitats with methanotrophic activity will help reveal whether the small εH value observed is a ubiquitous feature across all marine systems.

Carbon and hydrogen isotope fractionation during aerobic oxidation of short-chain alkanes in experimental incubations of vent fluids

Aerobic oxidation of short-chain alkanes was observed in gas samples from the Lutao intertidal hydrothermal vents in Taiwan, during storage at 20 °C for up to 29 months without adding bacterial strains and replenishing substrates. The carbon isotope fractionation factors (εC) of methane (C1), ethane (C2), and propane (C3), were calculated using the Rayleigh fractionation equation to be −37.1 ± 7.5‰, −14.8 ± 4.8‰, and −4.7 ± 5.2‰, respectively. The hydrogen isotope fractionation factor (εH) of methane was determined to be −281 ± 187‰. DNA sequencing of the 16S rRNA gene in the vent fluids suggests that aerobic oxidation is dominated by methanotrophs of the genera Methylomicrobium and Methylophaga, which use the ribulose monophosphate pathway (RuMP). The degrees of isotope fractionation (εC and εH values) herein are larger than previously reported values, possibly due to the limited O2 supply and low abundance of aerobic methane-oxidizing bacteria in the experiments. Since the fractionation factor of methane is higher than those of ethane and propane, the aerobic oxidation of thermogenic or microbial alkanes could produce a carbon isotope reversal, which is frequently noted as a trait of abiotic hydrocarbons. This work demonstrates that in addition to anaerobic microbial oxidation, aerobic oxidation with a low cell density can also produce significant isotope fractionation of alkanes in geological closed/semi-closed environments and open flow reaction systems that are characterized by moderate temperatures and a limited supply of substrates and O2; these environments include cold seeps, mud volcanoes, and low-temperature hydrothermal plumes/aquifers/reservoirs.

Origin and Source Characterization of Methane in the Shallow-Water Environment of Southern Lake Tanganyika Rift Basin, Tanzania

Lake Tanganyika, located in the western part of Tanzania between 3°S and 9°S lat, harbours hydrocarbon and non-hydrocarbon gases in its northern and southern shallow-water environment. In this study, a geochemical analysis of stable carbon and hydrogen isotopes and an interpretation of individual gas molecular composition were done in order to determine the origin and composition of the naturally occurring hydrocarbon gases in Tanganyika Basin. Nitrogen, a non-hydrocarbon gas is a major component (76.69%–78.31%) by volume percentage followed by methane (11.68%–12.94%) and other higher hydrocarbons (0.16%–1.63%). The isotopic composition of carbon δ 13 C 1 and hydrogen δ DC 1 ranges from –65.32‰ to –65.81‰ and –272.5‰ to –275.9‰ respectively. The isotopic compositions of ethane (δ DC 2 = –36.7‰ to –35.2‰) and propane (δ DC 3 = –31.3‰ to –27.5‰) reflect the thermogenic origin of these higher hydrocarbons. According to molecular characterization of carbon and hydrogen isotope ratios and δ D-values, methane gas falls in the biogenic origin category and is formed by carbon dioxide reduction. The isotopic composition of CO 2 varies between –8.6‰ and –3.4‰. CO 2 reduction is also regarded as a mechanism of biogenic methane formation based on carbon isotope fractionation factors (greater than 0.16).

Theoretical calculation of position-specific carbon and hydrogen isotope equilibriums in butane isomers

Position-specific isotope analysis has shown its potential to reveal information regarding formation, migration, and conversion procesess of hydrocarbons. The intramolecular isotope compositions of butane are promising to serve as a new thermometer and tracer. Therefore, position-specific isotope signatures in butane at equilibrium are needed for calibrating experimental measurements, establishing new geothermometers, and recognizing kinetic isotope effects. Here we conduct quantum chemistry modeling with corrections beyond the harmonic approximation and the Born-Oppenheimer approximation to obtain accurate intramolecular and intermolecular carbon and hydrogen isotope fractionation factors for butane isomers at equilibrium. Temperature dependences of these equilibrium isotope effects are presented for the range from 0 °C to 726.85 °C. The contribution of higher-order energy terms to 1000lnα values beyond the Bigeleisen-Mayer equation is found to be comparable to the magnitude of current experimental precisions. In addition to the significance of anharmonicity, the contribution of hindered internal rotation and diagonal Born-Oppenheimer correction is found to be important for accurate predictions of position-specific hydrogen isotope equilibriums. The abundance ratio of n-butane to i-butane at equilibriums is also calculated at various temperatures. Our results provide fundamental understanding of equilibrium properties for studying position-specific isotope effects in butane.

Lower hydrogen flux leads to larger carbon isotopic fractionation of methane and biomarkers during hydrogenotrophic methanogenesis

The isotopic composition of lipid biomarkers and biomass in sedimentary environments are widely used to infer microbial metabolisms and constrain carbon cycling processes. It has been observed that metabolic energy availability in the form of H2 impacts the stable carbon isotopes of CH4 during hydrogenotrophic methanogenesis, but it is unknown whether this relationship extends to lipids and amino acids. Since lipids and amino acids are long-lived, they can be used to reconstruct past conditions over geologic timescales. Therefore, the controls on their isotopic signatures are important to constrain to better interpret past environments.In this study, the isotopic distributions of carbon metabolized and reduced by the hyperthermophile Methanocaldococcus jannaschii were quantified following growth at 82 °C in a chemostat with high (80–83 µM) and low (15–27 µM) H2 concentrations. As has been shown previously, the stable carbon isotope fractionation factors for CH4 were >15‰ larger in low H2 experiments than in high H2 experiments. Lipid biomarkers and amino acids were similarly impacted with approximately 10‰ larger fractionation factors under low H2 conditions. The increase in fractionation factors can be related to the lower availability of thermodynamic energy, suggesting that even larger fractionation factors would be observed in methanogens living close to their threshold energy needs, as they do in most environments. The resulting isotopic signatures of long-lived lipid biomarkers synthesized by hydrogenotrophic methanogens may become as ‘superlight’ as those synthesized by archaea carrying out the anaerobic oxidation of methane. These results help to describe the underlying mechanisms that determine the isotopic composition of long-lived biomarkers and provide constraints for interpreting these signatures in the environment.

Experimental carbonatite/graphite carbon isotope fractionation and carbonate/graphite geothermometry

Abstract Carbon isotope exchange between carbon-bearing high temperature phases records the carbon (re-) processing in the Earth's interior, where the vast majority of global carbon is stored. Redox reactions between carbonate phases and elemental carbon govern the mobility of carbon, which then can be traced by its isotopes. We determined the carbon isotope fractionation factor between graphite and a Na2CO3-CaCO3 melt at 900–1500 °C and 1 GPa; The failure to isotopically equilibrate preexisting graphite led us to synthesize graphite anew from organic material during the melting of the carbonate mixture. Graphite growth proceeds by (1) decomposition of organic material into globular amorphous carbon, (2) restructuring into nano-crystalline graphite, and (3) recrystallization into hexagonal graphite flakes. Each transition is accompanied by carbon isotope exchange with the carbonate melt. High-temperature (1200–1500 °C) equilibrium isotope fractionation with type (3) graphite can be described by Δ 13 C c a r b o n a t e - g r a p h i t e = 3.17 ( ± 0.07 ) · 10 6 T 2 (temperature T in K). As the experiments do not yield equilibrated bulk graphite at lower temperatures, we combined the ≥1200 °C experimental data with those derived from upper amphibolite and lower granulite facies carbonate-graphite pairs (Kitchen and Valley, 1995; Valley and O'Neil, 1981). This yields the general fractionation function Δ 13 C c a r b o n a t e - g r a p h i t e = 3.37 ± 0.04 · 10 6 T 2 usable as a geothermometer for solid or liquid carbonate at ≥600 °C. Similar to previous observations, lower-temperature experiments (≤1100 °C) deviate from equilibrium. By comparing our results to diffusion and growth rates in graphite, we show that at ≤1100 °C carbon diffusion is slower than graphite growth, hence equilibrium surface isotope effects govern isotope fractionation between graphite and carbonate melt and determine the isotopic composition of newly formed graphite. The competition between diffusive isotope exchange and growth rates requires a more careful interpretation of isotope zoning in graphite and diamond. Based on graphite crystallization rates and bulk isotope equilibration, a minimum diffusivity of Dgraphite = 2 × 10−17 m2s−1 for T > 1150 °C is required. This value is significantly higher than calculated from experimental carbon self-diffusion constants (∼1.6 × 10−29 m2 s−1) but in good agreement with the value calculated for mono-vacancy migration (∼2.8 × 10−16 m2 s−1).

Aerobic microbial oxidation of hydrocarbon gases: Implications for oil and gas exploration

Abstract The molecular and carbon and hydrogen isotope compositions associated with the aerobic consumption of methane (Cl), ethane (C2) and propane (C3) were determined from incubations of marine sediments collected from the seafloor surface of the gas- and oil-bearing Bohai Bay in eastern China. The fresh sediment samples were incubated aerobically at 28 °C and treated with three types of hydrocarbon gas mixtures as follows: mixture I (C1:C2:C3 = 100:0:0), II (C1:C2:C3 = 99.00:0.70:0.30) and III (C1:C2:C3 = 79.96:10.04:10.00), corresponding to incubations i, ii and iii, respectively. All the hydrocarbons are of thermogenic origin, except C1 (microbial) in incubation ii. The oxidation of C1-C3 was observed in all three treatments, with variations in carbon isotope fractionation factors (eC) of C1 and average values of −10.5 ± 0.1, −5.7 ± 1.6, and −10.9 ± 2.9, respectively. The hydrogen isotope fractionation factors (eH) of C1 in the three incubations also display variation, with average values of −118.4 ± 3.0, −68.3 ± 15.0 and −56.3 ± 8.4, respectively. Experiments with thermogenic C1 display greater carbon fractionation than those with microbial C1. The eC and eH values for C2 and C3 were obtained only in incubation iii. Compared to C1, C2 and C3 exhibit weaker carbon isotope fractionation but stronger hydrogen isotope fractionation. The fractionation factors presented in this study can be applied to estimate the extent of C1-C3 oxidation in natural environments, especially at seafloor and terrestrial seeps where aerobic microbes are commonly present. Our study has important implications in oil and gas exploration, with an alert for exercising caution when using the molecular and isotopic characteristics of hydrocarbon gases to evaluate the gas sources because these are susceptible to aerobic microbial oxidation.

Quantification of Methanogenic Pathways Using Stable Carbon Isotopic Signatures.

In many anaerobic environments methane (CH4) is produced by methanogens, with either H2/CO2 or acetate (i.e., the methyl group) as precursors, through what are referred to as hydrogenotrophic and acetoclastic methanogenic pathways respectively. Their relative contribution to total CH4 production can be quantified by determining the stable carbon isotopic fractionation factors for both pathways as well as the isotopic signatures of CO2, CH4, and the methyl group in acetate of the sample. The procedures for measuring carbon isotopic fractionation factors of both methanogenic pathways and isotopic composition of these compounds by isotope ratio mass spectrometry are described in this chapter. The results are very helpful in evaluating the activity of the methanogens involved in each methanogenic pathway as well as those of other biological pathways with different fractionation factors.

Anaerobic methane oxidation by nitrate: kinetic isotope effect

The ratio of stable carbon isotopes (13C/12C) in different environments serves as a significant limitation in estimating the global balance of methane [Hornibrook et al., 2000]. In this case, the value of 13C/12C largely depends on the kinetic isotope effect associated with the metabolism of microorganisms that produce and consume CH4. The article suggests a dynamic model of the processes of methane formation and its anaerobic oxidation with nitrate by methanotrophic denitrifying microorganisms (DAOM), which allowed estimating the fractionation factor of stable carbon isotopes. In the experiment with peat from the minerotrophic bog [Smemo, Yavitt, 2007], the dynamics of the amount of methane and was measured. The dynamic model showed that the introduction of nitrate leads to a slow decrease in the partial pressure of methane. Since methane in the DAOM process is a substrate, methane is enriched with heavier carbon 13C in the system under study. This leads to an increase in the value . The carbon isotope fractionation factor during methane oxidation with nitrate was equal to 1.018 and comparable with the fraction of carbon isotope fractionation in the process of acetoclastic methanogenesis (1.01). Model calculations have shown that during incubation the apparent fractionation factor of carbon isotopes with the simultaneous formation of methane and DAOM slowly decreases. The ratio of 13C/12C isotopes in dissolved and gaseous methane practically does not differ. The model showed that an increase in the initial concentration of nitrate increases the rate of DAOM, which leads to a decrease in the concentration of dissolved methane. In this case, the value of 13C/12C increases. In field studies, Shi et al. (2017) showed that the presence of DAOM in peat bogs in which fertilizers penetrate can be controlled by the amount of nitrate used and the depth of penetration into the anoxic layer. Two MATLAB files describing DAOM are attached to the article.