Carbon Isotope Fractionation Factors Research Articles

Different types of methane monooxygenases produce similar carbon and hydrogen isotope fractionation patterns during methane oxidation

We determined the stable carbon and hydrogen isotope fractionation factors for methane oxidation under oxic conditions using strains with known degradation pathways. The aerobic oxidation of methane can be initiated by two different forms of enzymes known as methane monooxygenases (MMO). The expression of these enzymes is type-specific and dependent upon the adjusted copper concentration in the medium (or environment). In this study, the expression of either the soluble MMO or the particulate MMO was supported by adjusting the copper concentrations in the growth medium. Taxonomically different aerobic methanotrophic strains, mainly belonging to the alpha- and gamma- classes of Proteobacteria, produced methane isotope enrichment factors ( ε bulk) ranging from −14.8 to −27.9‰ for carbon, and from −110.0 to −231.5‰ for hydrogen. The ratio of hydrogen versus carbon discrimination ( Λ = ( α H −1 − 1)/( α C −1 − 1) ≈ Δ( δ 2H)/Δ( δ 13C)) were similar for all tested cultures, and are also identical to values calculated from previously published enrichment factors for aerobic and anaerobic methane degradation. In contrast, Λ-values for the abiotic oxidation of methane with OH radicals (this process is considered as the main removal process for methane from the atmosphere) were significantly higher than the values derived from biotic oxidation. Due to the low variability of microbial methane isotope fractionation patterns, we propose that combined carbon and hydrogen isotope fractionation analyses can be used to monitor and assess the occurrence of microbial methane oxidation in marine or terrestrial environments. However, it is not possible to distinguish distinct aerobic or anaerobic methane-oxidation pathways by this approach.

Insights into Enzyme Kinetics of Chloroethane Biodegradation Using Compound Specific Stable Isotopes

While compound specific isotope analysis (CSIA) has been used extensively to investigate remediation of chlorinated ethenes, to date considerably less information is available on its applicability to chlorinated ethanes. In this study, biodegradation of 1,1,1-trichloroethane (1,1,1-TCA) and 1,1-dichloroethane (1,1-DCA) was carried out by a Dehalobacter-containing mixed culture. Carbon isotope fractionation factors (ε) measured during whole cell degradation demonstrated that values for 1,1,1-TCA and 1,1-DCA (-1.8‰ and -10.5‰, respectively) were significantly smaller than values reported for abiotic reductive dechlorination of these same compounds. Similar results were found in experiments degrading these two priority pollutants by cell free extracts (CFE) where values of -0.8‰ and -7.9‰, respectively, were observed. For 1,1,1-TCA in particular, the large kinetic isotope effect expected for cleavage of a C-Cl bond was almost completely masked during biodegradation by both whole cells and CFE. Comparison to previous studies demonstrates that these patterns of isotopic fractionation are not attributable to transport effects across the cell membrane, as had been seen for other compounds such as PCE. In contrast these results reflect significant differences in the kinetics of the enzymes catalyzing chlorinated ethane degradation.

Assessment of MTBE biodegradation pathways by two-dimensional isotope analysis in mixed bacterial consortia under different redox conditions

The fuel oxygenate, methyl tert-butyl ether (MTBE), although now widely banned or substituted, remains a persistent groundwater contaminant. Multidimensional compound-specific isotope analysis (CSIA) of carbon and hydrogen is being developed for determining the extent of MTBE loss due to biodegradation and can also potentially distinguish between different biodegradation pathways. Carbon and hydrogen isotopic fractionation factors were determined for MTBE degradation in aerobic and anaerobic laboratory cultures. The carbon isotopic enrichment factor (epsilonC) for aerobic MTBE degradation by a bacterial consortium containing the aerobic MTBE-degrading bacterium, Variovorax paradoxus, was -1.1 +/- 0.2 per thousand and the hydrogen isotope enrichment factor (epsilonH) was -15 +/- 2 per thousand. This corresponds to an approximated lambda value (Lambda = epsilonH/epsilonC) of 14. Carbon isotope enrichment factors for anaerobic MTBE-degrading enrichment cultures were -7.0 +/- 0.2 per thousand and did not vary based on the original inoculum source, redox condition of the enrichment, or supplementation with syringic acid as a co-substrate. The hydrogen enrichment factors of cultures without syringic acid were insignificant, however a strong hydrogen enrichment factor of -41 +/- 3 per thousand was observed for cultures which were fed syringic acid during MTBE degradation. The Lambda = 6 obtained for NYsyr cultures might be diagnostic for the stimulation of anaerobic MTBE degradation by methoxylated compounds by an as yet unknown pathway and mechanism. The stable-isotope enrichment factors determined in this study will enhance the use of CSIA for monitoring anaerobic and aerobic MTBE biodegradation in situ.

Characterization of anaerobic xylene biodegradation by two‐dimensional isotope fractionation analysis

We determined stable carbon and hydrogen isotope fractionation factors for anaerobic degradation of xylene isomers by several pure and mixed cultures. All cultures initiated xylene degradation by the addition of fumarate to a methyl moiety, as is known from the literature or verified by the presence of methylbenzylsuccinates as metabolic intermediates. Additionally, the A subunit of benzylsuccinate synthase (bssA) was identified in the majority of the cultures by bssA-targeted primers. Xylene degradation was always coupled to a significant carbon and hydrogen isotope fractionation. The values of the apparent kinetic isotope effect (AKIE) for carbon and hydrogen indicate that the cleavage of a carbon-hydrogen bond is an isotope-sensitive step during fumarate addition to xylene isomers. The slopes of the linear regression for hydrogen (Δδ(2) H) versus carbon (Δδ(13) C) discrimination (Λ = Δδ(2) H/Δδ(13) C ≈ εHbulk /εCbulk ) ranged from 12 ± 4 to 29 ± 5 and were comparable to Λ values previously determined for anaerobic toluene degradation. The results suggest that combined carbon and hydrogen isotope fractionation analyses can be used to monitor anaerobic xylene degradation at contaminated sites.

Stable Isotope Fractionation of γ-Hexachlorocyclohexane (Lindane) during Reductive Dechlorination by Two Strains of Sulfate-Reducing Bacteria

Carbon isotope fractionation factors were determined with the dichloro elimination of gamma-hexachlorocyclohexane (gamma-HCH) by the sulfate-reducing bacteria Desulfococcus multivorans DSM 2059 and Desulfovibrio gigas DSM 1382. Both strains are known for cometabolic HCH dechlorination. Degradation experiments with gamma-HCH in concentrations of 22-25 gammaM were carried out using benzoate (for D. multivorans) and lactate (for D. gigas) as electron donors, respectively. Gamma-HCH was dechlorinated by both bacterial strains within four weeks, and the metabolites gamma-3,4,5,6-tetrachlorocyclohexene (gamma-TCCH), chlorobenzene (CB), and benzene were formed. The carbon isotope fractionation of gamma-HCH dechlorination was quantified by the Rayleigh model, using a bulk enrichment factor (epsilon C) of -3.9 +/- 0.6 for D. gigas and -3.4 +/- 0.5 for D. multivorans, which correspond to apparent kinetic isotope effect (AKIEc) values of 1.023 +/- 0.004 or 1.02 +/- 0.003 for stepwise Cl-C bond cleavage. The extent and range of isotope fractionation suggest that gamma-HCH dechlorination can be monitored in anoxic environments by compound-specific isotope analysis (CSIA).

Calculation of site-specific carbon-isotope fractionation in pedogenic oxide minerals

Ab initio molecular dynamics and quantum chemistry techniques are used to calculate the structure, vibrational frequencies, and carbon-isotope fractionation factors of the carbon dioxide component [CO(2)(m)] of soil (oxy)hydroxide minerals goethite, diaspore, and gibbsite. We have identified two possible pathways of incorporation of CO(2)(m) into (oxy)hydroxide crystal structures: one in which the C(4+) substitutes for four H(+) [CO(2)(m)(A)] and another in which C(4+) substitutes for (Al(3+),Fe(3+)) + H(+) [CO(2)(m)(B)]. Calculations of isotope fractionation factors give large differences between the two structures, with the CO(2)(m)(A) being isotopically lighter than CO(2)(m)(B) by approximately 10 per mil in the case of gibbsite and nearly 20 per mil in the case of goethite. The reduced partition function ratio of CO(2)(m)(B) structure in goethite differs from CO(2)(g) by <1 per mil. The predicted fractionation for gibbsite is >10 per mil higher, close to those measured for calcite and aragonite. The surprisingly large difference in the carbon-isotope fractionation factor between the CO(2)(m)(A) and CO(2)(m)(B) structures within a given mineral suggests that the isotopic signatures of soil (oxy)hydroxide could be heterogeneous.

Distribution of lipid biomarkers and carbon isotope fractionation in contrasting trophic environments of the South East Pacific

Abstract. The distribution of lipid biomarkers and their stable carbon isotope composition was investigated on suspended particles from different contrasting trophic environments at six sites in the South East Pacific. High algal biomass with diatom-related lipids (24-methylcholesta-5,24(28)-dien-3β-ol, C25 HBI alkenes, C16:4 FA, C20:5 FA) was characteristic in the upwelling zone, whereas haptophyte lipids (long-chain (C37-C39) unsaturated ketones) were proportionally most abundant in the nutrient-poor settings of the centre of the South Pacific Gyre and on its easter edge. The dinoflagellate–sterol, 4α-23,24-trimethylcholest-22(E)-en-3β-ol, was a minor contributor in all of the studied area and the cyanobacteria-hydrocarbon, C17n-alkane, was at maximum in the high nutrient low chlorophyll regime of the subequatorial waters near the Marquesas archipelago. The taxonomic and spatial variability of the relationships between carbon photosynthetic fractionation and environmental conditions for four specific algal taxa (diatoms, haptophytes, dinoflagellates and cyanobacteria) was also investigated. The carbon isotope fractionation factor (εp) of the 24-methylcholesta-5,24(28)-dien-3β-ol diatom marker, varied over a range of 16% along the different trophic systems. In contrast, εp of dinoflagellate, cyanobacteria and alkenone markers varied only by 7–10‰. The low fractionation factors and small variations between the different phytoplankton markers measured in the upwelling area likely reveals uniformly high specific growth rates within the four phytoplankton taxa, and/or that transport of inorganic carbon into phytoplankton cells may not only occur by diffusion but also by other carbon concentrating mechanisms (CCM). In contrast, in the oligotrophic zone, i.e. gyre and eastgyre, relatively high εp values, especially for the diatom marker, indicate diffusive CO2 uptake by the eukaryotic phytoplankton. At these nutrient-poor sites, the lower εp values for haptophytes, dinoflagellates and cyanobacteria indicate higher growth rates or major differences on the carbon uptake mechanisms compared to diatoms.

Assessment of microbial methane oxidation above a petroleum‐contaminated aquifer using a combination of in situ techniques

Emissions of the greenhouse gas CH4, which is often produced in contaminated aquifers, are reduced or eliminated by microbial CH4 oxidation in the overlying vadose zone. The aim of this field study was to estimate kinetic parameters and isotope fractionation factors for CH4 oxidation in situ in the vadose zone above a methanogenic aquifer in Studen, Switzerland, and to characterize the involved methanotrophic communities. To quantify kinetic parameters, several field tests, so‐called gas push‐pull tests (GPPTs), with CH4 injection concentrations ranging from 17 to 80 mL L−1 were performed. An apparent Vmax of 0.70 ± 0.15 mmol CH4 (L soil air)−1 h−1 and an apparent Km of 0.28 ± 0.09 mmol CH4 (L soil air)−1 was estimated for CH4 oxidation at 2.7 m depth, close to the groundwater table. At 1.1 m depth, Km (0.13 ± 0.02 mmol CH4 (L soil air)−1) was in a similar range, but Vmax (0.076 ± 0.006 mmol CH4 (L soil air)−1 h−1) was an order of magnitude lower. At 2.7 m, apparent first‐order rate constants determined from a CH4 gas profile (1.9 h−1) and from a single GPPT (2.0 ± 0.03 h−1) were in good agreement. Above the groundwater table, a Vmax much higher than the in situ CH4 oxidation rate prior to GPPTs indicated a high buffer capacity for CH4. At both depths, known methanotrophic species affiliated with Methylosarcina and Methylocystis were detected by cloning and sequencing. Apparent stable carbon isotope fractionation factors α for CH4 oxidation determined during GPPTs ranged from 1.006 to 1.032. Variability was likely due to differences in methanotrophic activity and CH4 availability leading to different degrees of mass transfer limitation. This complicates the use of stable isotopes as an independent quantification method.

Quantum-Chemical Calculations of Carbon-Isotope Fractionation in CO2(g), Aqueous Carbonate Species, and Carbonate Minerals

Quantum chemical calculations on large supermolecular carbonate-water and carbonate mineral clusters are used to predict equilibrium constants for 13,12C-isotope-exchange reactions between CO2(g), aqueous carbonate species, and the common carbonate minerals. For the aqueous species, we evaluate the influence of the size and conformational variability of the solvation shell, the exchange-correlation functional, and the basis set. The choice of exchange-correlation functional (PBE vs B3LYP), the basis set (6-31G* vs aug-cc-pVDZ), and solvation shell size (first shell only vs first shell and a partial second shell) each produce changes of approximately 5-10 per mil in the reduced partition function ratio. Conformational variability gives rise to a standard error of approximately 0.5 per mil using approximately 10 solute-solvent conformations. The best results are obtained with the B3LYP/aug-cc-pVDZ combination, but because the improvements in the basis set and exchange correlation functional drive the reduced partition function ratios in opposite directions, reasonably good results are also obtained with the PBE/6-31G* combination. To construct molecular clusters representative of mineral environments, a new method is introduced on the basis of conservation of Pauling bond strength. Using these clusters as models for minerals, calculations of mineral-gas and mineral-aqueous carbon-isotope fractionation factors, are in good agreement with experimental measurements. Carbon-isotope fractionation factors for gas, aqueous, and mineral phases are thus integrated into a single theoretical/computational framework.

Methane Oxidation in Biofilters Measured by Mass-Balance and Stable Isotope Methods

Simultaneous flux and isotope measurements on compost and sand biofilters showed that the fraction of CH4 oxidized, calculated from delta13C measurements using a closed system model (f(oxir,C)), averaged only 0.455 of the fraction oxidized based on mass-balance measurements (f(oxm)). The discrepancy between f(oxm) and f(oxir,C) may be partly due to complete oxidation of a portion of the inflow gas, thereby eliminating its contribution to the emitted methane on which isotopic measurements are conducted. To relate f(oxir,C) and f(oxm) a simple binary closed-system model is proposed that assumes that f(oxir,C) refers to only part of the inflow, P, and that the remainder of inflow (1 - P) is completely oxidized before reaching the outlet. This model is compared to the standard open-system model. The H-isotope fraction oxidized (f(oxir,H)) was determined for a subset of samples and found to be not significantly different from f(oxir,C). The carbon isotope fractionation factor, alphaox,C = 1.0244, and the H-isotope fractionation factor, alphaox,H = 1.2370, were determined by incubation studies. Delta13C measurements indicated that the emitted flow was more strongly oxidized by the compost biofilters (f(oxir,C) = 0.362, f(oxm) = 0.757) than the sand biofilters (f(oxir,C) = 0.222, f(oxm) = 0.609).

Carbon and hydrogen isotope fractionation associated with the aerobic microbial oxidation of methane, ethane, propane and butane

Carbon isotope fractionation factors associated with the aerobic consumption of methane (C1), ethane (C2), propane (C3), and n-butane (C4) were determined from incubations of marine sediment collected from the Coal Oil Point hydrocarbon seep field, located offshore Santa Barbara, CA. Hydrogen isotope fractionation factors for C1, C2 and C3 were determined concurrently. Fresh sediment samples from two seep areas were each slurried with sea water and treated with C1, C2, C3 or C4, or with mixtures of all four gases. Triplicate samples were incubated aerobically at 15 °C, and the stable isotope composition and headspace levels of C1–C4 were monitored over the course of the experiment. Oxidation was observed for all C1–C4 gases, with an apparent preference for C3 and C4 over C1 and C2 in the mixed-gas treatments. Fractionation factors were calculated using a Rayleigh model by comparing the δ 13C and δD of the residual C1–C4 gases to their headspace levels. Carbon isotope fractionation factors (reported in ε or ( α-1) × 1000 notation) were consistent between seep areas and were −26.5‰ ± 3.9 for C1, −8.0‰ ± 1.7 for C2, −4.8‰ ± 0.9 for C3 and −2.9‰ ± 0.9 for C4. Fractionation factors determined from mixed gas incubations were similar to those determined from individual gas incubations, though greater variability was observed during C1 consumption. In the case of C1 and C3 consumption, carbon isotope fractionation appears to decrease as substrate becomes limiting. Hydrogen isotope fractionation factors determined from the two seep areas differed for C1 oxidation but were similar for C2 and C3. Hydrogen isotope fractionation factors ranged from −319.9‰ to −156.4‰ for C1 incubations, and averaged −61.9‰ ± 8.3 for C2 incubations and −15.1‰ ± 1.9 for C3 incubations. The fractionation factors presented here may be applied to estimate the extent of C1–C4 oxidation in natural gas samples, and should prove useful in further studying the microbial oxidation of these compounds in the natural environment.

Preferential formation of 13C– 18O bonds in carbonate minerals, estimated using first-principles lattice dynamics

Equilibrium constants for internal isotopic exchange reactions of the type: Ca 12 C 18 O 16 O 2 + Ca 13 C 16 O 3 ↔ Ca 13 C 18 O 16 O 2 + Ca 12 C 16 O 3 for individual CO 3 2− groups in the carbonate minerals calcite (CaCO 3), aragonite (CaCO 3), dolomite (CaMg(CO 3) 2), magnesite (MgCO 3), witherite (BaCO 3), and nahcolite (NaHCO 3) are calculated using first-principles lattice dynamics. Calculations rely on density functional perturbation theory (DFPT) with norm-conserving planewave pseudopotentials to determine the vibrational frequencies of isotopically substituted crystals. Our results predict an ∼0.4‰ excess of 13 C 18 O 16 O 2 2 − groups in all studied carbonate minerals at room-temperature equilibrium, relative to what would be expected in a stochastic mixture of carbonate isotopologues with the same bulk 13C/ 12C, 18O/ 16O, and 17O/ 16O ratios. The amount of excess 13 C 18 O 16 O 2 2 − decreases with increasing temperature of equilibration, from 0.5‰ at 0 °C to <0.1‰ at 300 °C, suggesting that measurements of multiply substituted isotopologues of carbonate could be used to infer temperatures of ancient carbonate mineral precipitation and alteration events, even where the δ 18O of coexisting fluids is uncertain. The predicted temperature sensitivity of the equilibrium constant is ∼0.003‰/°C at 25 °C. Estimated equilibrium constants for the formation of 13 C 18 O 16 O 2 2 − are remarkably uniform for the variety of minerals studied, suggesting that temperature calibrations will also be applicable to carbonate minerals not studied here without greatly compromising accuracy. A related equilibrium constant for the reaction: Ca 12 C 18 O 16 O 2 + Ca 12 C 17 O 16 O 2 ↔ Ca 12 C 18 O 17 O 16 O + Ca 12 C 16 O 3 in calcite indicates formation of 0.1‰ excess 12C 18O 17O 16O 2− at 25 °C. In a conventional phosphoric acid reaction of carbonate to form CO 2 for mass-spectrometric analysis, molecules derived from 13 C 18 O 16 O 2 2 − dominate (∼96%) the mass 47 signal, and 12C 18O 17O 16O 2− contributes most of the remainder (3%). This suggests that carbonate internal equilibration temperatures can be recovered from acid-generated CO 2 if abundances of isotopologues with mass 44–47 can be measured to sufficient precision. We have also calculated 18O/ 16O and 13C/ 12C reduced partition function ratios for carbonate minerals, and find them to be in good agreement with experiments and empirical calibrations. Carbon and oxygen isotope fractionation factors in hypothetical 40Mg—magnesite and 40Ba—witherite indicate that M 2+-cation mass does not contribute significantly to equilibrium isotopic fractionations between carbonate minerals.

Variable carbon isotope fractionation expressed by aerobic CH 4-oxidizing bacteria

Carbon isotope fractionation factors reported for aerobic bacterial oxidation of CH 4 ( α CH 4 – CO 2 ) range from 1.003 to 1.039. In a series of experiments designed to monitor changes in the carbon isotopic fractionation of CH 4 by Type I and Type II methanotrophic bacteria, we found that the magnitude of fractionation was largely due to the first oxidation step catalyzed by methane monooxygenase (MMO). The most important factor that modulates the ( α CH 4 – CH 3 OH ) is the fraction of the total CH 4 oxidized per unit time, which strongly correlates to the cell density of the growth cultures under constant flow conditions. At cell densities of less than 0.1 g/L, fractionation factors greater than 1.03 were observed, whereas at cell densities greater than 0.5 g/L the fractionation factors decreased to as low as 1.002. At low cell densities, low concentrations of MMO limit the amount of CH 4 oxidized, while at higher cell densities, the overall rates of CH 4 oxidation increase sufficiently that diffusion of CH 4 from the gaseous to dissolved state and into the cells is likely the rate-determining step. Thus, the residual CH 4 is more fractionated at low cell densities, when only a small fraction of the total CH 4 has been oxidized, than at high cell densities, when up to 40% of the influent CH 4 has been utilized. Therefore, since Rayleigh distillation behavior is not observed, δ 13C values of the residual CH 4 cannot be used to infer the amount oxidized in either laboratory or field-studies. The measured ( α CH 4 – CH 3 OH ) was the same for both Type I and Type II methanotrophs expressing particulate or soluble MMO. However, large differences in the δ 13C values of biomass produced by the two types of methanotrophs were observed. Methylosinus trichosporium OB3b (Type II) produced biomass with δ 13C values about 15‰ higher than the dissimilated CO 2, whereas Methylomonas methanica (Type I) produced biomass with δ 13C values only about 6‰ higher than the CO 2. These effects were independent of the magnitude of the initial carbon isotope fractionation caused by MMO and were relatively constant despite changing ratios of assimilatory to dissimilatory carbon transformation by the organisms. This suggests that the difference in biomass carbon isotopes is primarily due to differences in the fractionation effect at the formaldehyde branch point in the metabolic pathway, rather than assimilation of CO 2 by Type II methanotrophs.

Carbon isotope fractionation during degradation of benzene, trichloroethene, and tetrachloroethene under ultraviolet light

Stable carbon isotope fractionation factors associated with the degradation of benzene, trichloroethene (TCE), and tetrachloroethene (PCE) under ultraviolet light (254 nm) have been measured experimentally using simple and inexpensive quartz reaction vessels equipped with a syringe valve and septum. Carbon isotope enrichment factors of ebenzene = -6.2 ± 0.3‰, eTCE = -6.8 ± 0.2‰, and ePCE = -4.1 ± 0.2‰ were obtained. The results demonstrate that the simple and inexpensive experimental design used in this study has potential application to determine isotope enrichment factors associated with degradation under conditions more representative of important atmospheric processes, and hence has future application to the study of the behavior of important trace gases such as non-methane hydrocarbons and other volatile organic compounds in the atmosphere.

Carbon isotope fractionation in synthetic magnesian calcite

Mg-calcite was precipitated at 25 °C in closed system, free-drift experiments, from solutions containing NaHCO 3, CaCl 2 and MgCl 2. The carbon stable isotope composition of bulk solid and solution were analyzed from subsamples collected during time course experiments of 24 h duration. Considering only the Mg-content and δ 13C values for the bulk solid, the carbon isotope fractionation factor for the Mg-calcite–HCO 3(aq) − system (as 10 3 ln α Mg - cl - HCO 3 ( aq ) - ) increased with average mol percentage of Mg (X Mg) in the solid at a rate of (0.024 ± 0.011) per mol% MgCO 3. Extrapolation of this relationship to the pure calcite end member yields a value of 0.82 ± 0.09, which is similar to published values for the calcite–HCO 3(aq) − system. Although 10 3 ln α Mg - cl – HCO 3 ( aq ) - did not vary for precipitation rates that ranged from 10 3.21 to 10 4.60 μmol m −2 h −1, it was not possible to hold Mg-content of the solid constant, so kinetic effect on 10 3 ln α could not be evaluated from these experiments.

Isotopic Fractionation of Carbon and Nitrogen During the Illicit Processing of Cocaine and Heroin in South America

The forensic application of stable isotope analysis to cocaine and heroin for geolocation of exhibits must take into account the possible enrichment and/or depletion of 13C and 15N during the illicit manufacturing process. Continuous-flow elemental analysis-isotope ratio mass spectrometry was utilized to measure changes in the stable isotope ratios of carbon and nitrogen for both cocaine (N = 92) and heroin/morphine (N = 81) exhibits derived from illicit manufacturing processes utilized by South American clandestine chemists. In controlled settings in South America, there was no siginficiant carbon isotope fractionation during the conversion of cocaine base to cocaine HCI using current illict methodologies. In contrast, nitrogen isotope fractionation for this conversion was 1 per thousand. There was a kinetic carbon isotope ratio fractionation during the acetylation of Colombian morphine to heroin and as a result heroin exhibits will almost always have more negative delta13C values than the original morphine. There was an isotopic fractionation against 15N during the acetylation of morphine base to heroin base, but this effect was not expressed since all of the heroin base was precipitated during the manufacturing process. However, the clandestine process of converting a single batch of heroin base usually involved two consecutive crops of heroin HCl and the latter crop was isotopically depleted as expected from a Rayleigh distillation process. When heroin was deacetylated to morphine, the morphine produced resulted in delta13C values that were indistinguishable from the original morphine. The kinetic carbon isotope fractionation factor for the South American process of morphine acetylation was -1.8 per thousand, allowing calculation of the delta13C values of the acetic anhydride from deacetylated heroin delta13C values.

Stable carbon isotope fractionation during degradation of dichloromethane by methylotrophic bacteria

Stable carbon isotope fractionation during dichloromethane (DCM) degradation by methylotrophic bacteria was investigated under aerobic and nitrate-reducing conditions. The strains studied comprise several Hyphomicrobium strains, Methylobacterium, Methylopila, Methylophilus and Methylorhabdus spp. that are considered to degrade DCM by a glutathione (GSH)-dependent dehalogenase enzyme system in the initial step. The stable carbon isotope fractionation factors (alphaC) of the strains varied under aerobic conditions between 1.043 and 1.071 and under nitrate-reducing conditions between 1.048 and 1.065. Comparison of isotope fractionation under aerobic and nitrate-reducing conditions by individual strains revealed only minor to no differences. The variability in isotope fractionation among strains was found to be related to the polymorphism of the functional genes encoding the DCM dehalogenase.

Stable carbon isotope fractionation during aerobic biodegradation of chlorinated ethenes.

Stable isotope analysis is recognized as a powerful tool for monitoring, assessing, and validating in-situ bioremediation processes. In this study, kinetic carbon isotope fractionation factors (epsilon) associated with the aerobic biodegradation of vinyl chloride (VC), cis-1,2-dichloroethylene (cDCE), and trichloroethylene (TCE) were examined. Of the three solvents, the largest fractionation effects were observed for biodegradation of VC. Both metabolic and cometabolic VC degradation were studied using Mycobacterium aurum L1 (grown on VC), Methylosinus trichosporium OB3b (grown on methane), Mycobacterium vaccae JOB5 (grown on propane), and two VC enrichment cultures seeded from contaminated soils of Alameda Point and Travis Air Force Base, CA. M. aurum L1 caused the greatest fractionation (epsilon = -5.7) while for the cometabolic cultures, epsilon values ranged from -3.2 to -4.8. VC fractionation patterns for the enrichment cultures were within the range of those observed for the metabolic and cometabolic cultures (epsilon = -4.5 to -5.5). The fractionation for cometabolic degradation of TCE by Me. trichosporium OB3b was low (epsilon = -1.1), while no quantifiable carbon isotopic fractionation was observed during the cometabolic degradation of cDCE. For all three of the tested chlorinated ethenes, isotopic fractionation measured during aerobic degradation was significantly smaller than that reported for anaerobic reductive dechlorination. This study suggests that analysis of compound-specific isotopic fractionation could assist in determining whether aerobic or anaerobic degradation of VC and cDCE predominates in field applications of in-situ bioremediation. In contrast, isotopic fractionation effects associated with metabolic and cometabolic reactions are not sufficiently dissimilar to distinguish these processes in the field.

Precipitation kinetics and carbon isotope partitioning of inorganic siderite at 25°C and 1 atm

Siderite was precipitated from NaHCO 3 and Fe(ClO 4) 2 solutions under anaerobic conditions at 25°C and 1 atm total pressure using a modified version of the chemo-stat technique and the free-drift technique. Samples of solution and solid were withdrawn at different time intervals during time course experiments to determine the bulk and isotope composition of the solution and solid, and the morphology and mineralogy of the solid. A series of metastable precursors precipitated and dissolved sequentially, culminating in well-crystallized siderite rhombohedra having an average edge of ∼ 2 μm and a limited size distribution. Siderite precipitation rate ranged from 10 0.23 to 10 2.44 μmol•m −2•h −1 for saturation states (with respect to siderite) ranging from near equilibrium to 10 3.53. Calculated carbon isotope fractionation factors (10 3lnα) averaged 8.5 ± 0.2 (1σ n = 4) for the siderite-CO 2(g) system and 0.5 ± 0.2 (1σ n = 4) for the siderite-HCO 3 − (aq) system.

Carbon isotope fractionation during permanganate oxidation of chlorinated ethylenes (cDCE, TCE, PCE).

Permanganate oxidation of chlorinated ethylenes is an attractive technique to effect remediation of these important groundwater contaminants. Stable carbon isotope fractionation associated with permanganate oxidation of trichloroethylene (TCE), tetrachloroethylene (PCE), and cis-1,2-dichloroethylene (cDCE) has been measured, to study the possibility of applying stable carbon isotope analysis as a technique to assess the efficacy of remediation implemented by permanganate oxidation. Average carbon isotope fractionation factors of alphaTCE = 0.9786, alphaPCE = 0.9830, and alphacDCE = 0.9789 were obtained, although the fractionation factor for PCE may be interpreted to change from a value of 0.9779-0.9871 during the course of the reaction. The fractionation factors for all three compounds are quite similar, in contrast to the variation of fractionation factors vs degree of chlorination observed for other degradative processes, such as microbial dechlorination. This may be due to a common rate-determining step for permanganate oxidation of all three compounds studied. The large fractionation factors and the relative lack of dependence of the fractionation factors upon other environmental factors (e.g. oxidation rate, presence of multiple contaminants, incomplete oxidation, presence of chloride in solution) indicate that monitoring delta13C values of chlorinated ethylenes during oxidation with permanganate may be a sensitive, and potentially quantitative, technique to investigate the extent of degradation.