Kinetic Isotope Effect Research Articles

Kinetic Isotope Effects During Reduction of Fe(III) to Fe(II): Large Normal and Inverse Isotope Effects for Abiotic Reduction and Smaller Fractionations by Phytoplankton in Culture

AbstractIron stable isotopes (δ56Fe) are a useful tool for studying Earth processes, many of which involve redox transformations between Fe(III) and Fe(II). Here, we present two related experimental efforts, a study of the kinetic isotope effects (KIEs) associated with the reduction of Fe(III)‐ethylenediaminetetraacetic acid (EDTA) to Fe(II), and measurements of the biological fractionation of Fe isotopes by phytoplankton in culture. Reductants tested were ascorbate, hydroxylamine, Mn(II), dithionite, and photoreduction at pH between 5 and 9 and temperatures from 0 to 100°C. Isotope fractionations were very large, and included both normal and inverse KIEs, ranging from −4‰ to +5‰. Experiments were reproducible, yielding similar results for triplicate experiments run concurrently and for experiments run weeks apart. However, fractionations were not predictable, without a clear relationship to reaction rate, temperature, pH, or the reductant used. Acquisition of Fe by eukaryotic phytoplankton also often involves the reduction of Fe(III) to Fe(II). Several species of diatoms and a coccolithophore were tested for Fe isotope fractionation in culture using EDTA, NTA, and DFB as Fe(III) chelating ligands, yielding fractionations from −1.3‰ to +0.6‰. Biological isotope effects were also unpredictable, showing no clear relationship to species, growth rate, or Fe concentration. Variability in Fe isotope fractionation observed in culture may be explained in part by the sensitivity of KIEs. This work has implications for the industrial purification of isotopes, interpretation of natural δ56Fe, and the use of Fe isotopes as a tracer Fe source and biological processes in the ocean and other natural systems.

Hydrogen isotope effect on thermodynamic and kinetic properties of Pd based ternary alloys: Pd–Ag-M (M = Au, Zr)

Owing to the importance of hydrogen and its isotopes in energy sector, the present study investigates thermodynamic and kinetic hydrogen isotope effects on desorption reaction of newly synthesized Pd-based ternary alloy-hydrides/deuterides viz., Pd0.89Ag0.05M0.06 (M = Au, Zr)–H/D. Alloys were prepared by arc melting method followed by characterization using PXRD, SEM, TXRF and EDX techniques. The effect of hydrogen isotopes on thermodynamics of H2/D2 desorption reaction was investigated by generating pressure-temperature isotherms (PCIs) at various temperatures using a Sieverts' type volumetric apparatus. The PCT diagram of alloys depicts a single plateau which confirms desorption to be a one-step process. Both alloy systems display a normal isotope effect as indicated by higher plateau pressure for deuterides as compared to hydrides. The enthalpy and entropy changes for the desorption reactions were calculated using Van't Hoff equation. The separation factor derived from PCT data indicates that Pd0.89Ag0.05Au0.06 alloy has a significantly high separation factor as compared to other Pd based alloys reported in the literature. To get more insight into this behavior, non-isothermal kinetic analysis of H2/D2 desorption based on Differential Scanning Calorimeter (DSC) data acquired at four different heating rates was conducted and activation energies of desorption were calculated by invoking Kissinger's equation. The activation energies of alloys were found to be lower than that of palladium which explains the high separation factor of alloys as compared to Pd. The desired combination of higher separation factor and lower activation energy makes Pd0.89Ag0.05Au0.06 alloy a promising material for practical application in the separation of hydrogen isotopes via self-displacement gas chromatography (SDGC).

Resolving and correcting for kinetic biases on methane seep paleotemperature using carbonate ∆47/∆48 analysis.

Methane-derived authigenic carbonate often constitutes the sole remaining record of relic methane seeps. The clumped (∆47) and oxygen isotopic composition of seep carbonates often yield inaccurate temperatures, attributed to kinetic isotope effects and modification of seawater isotope composition by hydrate water. Here, we analyzed the dual-clumped isotope (∆47/∆48) composition of authigenic carbonate from a modern methane seep. We demonstrate that aragonite forms closest to isotopic equilibrium such that its ∆47 can directly yield the correct formational temperature, whereas calcite is unambiguously biased by kinetic isotope effects. Numerical models show that the observed bias in the isotopic composition arises from rate-limiting dehydration/dehydroxylation of HCO3- alongside diffusive fractionation, which can be corrected for with analysis of carbonate ∆47/∆48 values. We demonstrate the utility of dual-clumped isotope analysis for studying seep carbonates, as it reveals the origin and magnitude of kinetic biases and can be used to reconstruct paleotemperature and seawater δ18O.

Unraveling the Oxidation Mechanism of Formic Acid on Pd(111) Electrode: Implication from pH Effect and H/D Kinetic Isotope Effect

Unraveling the Oxidation Mechanism of Formic Acid on Pd(111) Electrode: Implication from pH Effect and H/D Kinetic Isotope Effect

Hydrogen Tunneling Exhibiting Unexpectedly Small Primary Kinetic Isotope Effects.

Probing quantum mechanical tunneling (QMT) in chemical reactions is crucial to understanding and developing new transformations. Primary H/D kinetic isotopic effects (KIEs) beyond the semiclassical maximum values of 7-10 (room temperature) are commonly used to assess substantial QMT contributions in one-step hydrogen transfer reactions, because of the much greater QMT probability of protium vs. deuterium. Nevertheless, we report here the discovery of a reaction model occurring exclusively by H-atom QMT with residual primary H/D KIEs. 2-Hydroxyphenylnitrene, generated in N2 matrix, was found to isomerize to an imino-ketone via sequential (domino) QMT involving anti to syn OH-rotamerization (rate determining step) and [1,4]-H shift reactions. These sequential QMT transformations were also observed in the OD-deuterated sample, and unexpected primary H/D KIEs between 3 and 4 were measured at 3 to 20 K. Analogous residual primary H/D KIEs were found in the anti to syn OH-rotamerization QMT of 2-cyanophenol in a N2 matrix. Evidence strongly indicates that these intriguing isotope-insensitive QMT reactivities arise due to the solvation effects of the N2 matrix medium, putatively through coupling with the moving H/D tunneling particle. Should a similar scenario be extrapolated to conventional solution conditions, then QMT may have been overlooked in many chemical reactions.

Accessing Z‐Enynes via Cobalt‐Catalyzed Propargylic Dehydrogenation

AbstractAlkenes constitute an enabling motif in organic synthesis, as they can be functionalized to form highly substituted molecules. Z‐alkenes are generally challenging to access due to the thermodynamic preference for the formation of E‐alkenes compared to Z‐alkenes. Dehydrogenation methodologies to selectively form Z‐alkenes have not yet been reported. Herein, we report a Z‐selective, propargylic dehydrogenation that provides 1,3‐enynes through the invention of a Co‐catalyzed oxidation system. Observation of a kinetic isotope effect (KIE) revealed that deprotonation of the propargylic position is the rate limiting step. Additionally, isomerization experiments were conducted and confirmed that the observed Z‐selectivity is a kinetic effect. A proposed stereomechanistic model for the Z‐selectivity is included.

Accessing Z-Enynes via Cobalt-Catalyzed Propargylic Dehydrogenation.

Alkenes constitute an enabling motif in organic synthesis, as they can be functionalized to form highly substituted molecules. Z-alkenes are generally challenging to access due to the thermodynamic preference for the formation of E-alkenes compared to Z-alkenes. Dehydrogenation methodologies to selectively form Z-alkenes have not yet been reported. Herein, we report a Z-selective, propargylic dehydrogenation that provides 1,3-enynes through the invention of a Co-catalyzed oxidation system. Observation of a kinetic isotope effect (KIE) revealed that deprotonation of the propargylic position is the rate limiting step. Additionally, isomerization experiments were conducted and confirmed that the observed Z-selectivity is a kinetic effect. A proposed stereomechanistic model for the Z-selectivity is included.

High-Resolution Crossed-Beam Dynamics Studies of the D + Para-H2 → HD + H Reaction at 1.21 eV.

Understanding kinetic isotope effects is important in the study of the reaction dynamics of elementary chemical reactions, particularly those involving hydrogen atoms and molecules. As one of the isotopic variants of the hydrogen exchange reaction, the D + para-H2 reaction has attracted much attention. However, experimental studies of this reaction have been limited primarily due to its strong experimental background noise. In this study, by using the velocity map ion imaging method and the near-threshold ionization technique, together with improvements on the vacuum condition in the vicinity of the collision zone, background noise was reduced significantly, and quantum state-resolved differential cross sections (DCSs) for the D + para-H2 reaction at a collision energy of 1.21 eV were acquired in a crossed molecular beams experiment. Interestingly, clear rotational state-dependent angular distributions were noticed in the quantum state-resolved DCSs. The most intense peak's positions for HD (v', j') products shift to different scattering directions as the product's ro-vibrational quantum number increases. Two different microscopic reaction mechanisms are found to be involved in this reaction for HD products in different vibrational states. The results show a direct correlation between the scattering angle and the product's rotational quantum number, revealing that the contributions of impact parameters are strongly influenced by the corresponding centrifugal barrier.

Gold(III) Auracycles featuring C(sp3)−Au−C(sp2) Bonds: Synthesis and Mechanistic Insights into the Cycloauration Step

The direct auration of arenes is a key step in numerous gold‐catalyzed reactions. Although reported more than 100 years ago, understanding of its underlying mechanism has been hampered by the difficulties in the isolation of relevant intermediates given the propensity of gold(III) species to undergo reductive elimination. Here, we report the synthesis and isolation of a new family of intriguing zwitterionic [C(sp3)^C(sp2)]‐auracyclopentanes, as well as of their alkyl‐gold(III) precursors and demonstrate their value as mechanistic probes to study the Csp2‐Au bond‐forming event. Experimental investigations employing Kinetic Isotope Effects (KIE), Hammett plot, and Eyring analysis provided important insights into the formation of the auracycle. The data suggest a SEAr mechanism wherein the slowest step might be the p‐coordination between the arene and the gold(III) center, en route to the Wheland intermediate. We also show that these auracyclopentanes can work as catalysts in several gold‐promoted transformations.

Heavy-atom tunnelling in singlet oxygen deactivation predicted by instanton theory with branch-point singularities

The reactive singlet state of oxygen (O2) can decay to the triplet ground state nonradiatively in the presence of a solvent. There is a controversy about whether tunnelling is involved in this nonadiabatic spin-crossover process. Semiclassical instanton theory provides a reliable and practical computational method for elucidating the reaction mechanism and can account for nuclear quantum effects such as zero-point energy and multidimensional tunnelling. However, the previously developed instanton theory is not directly applicable to this system because of a branch-point singularity which appears in the flux correlation function. Here we derive a new instanton theory for cases dominated by the singularity, leading to a new picture of tunnelling in nonadiabatic processes. Together with multireference electronic-structure theory, this provides a rigorous framework based on first principles that we apply to calculate the decay rate of singlet oxygen in water. The results indicate a new reaction mechanism that is 27 orders of magnitude faster at room temperature than the classical process through the minimum-energy crossing point. We find significant heavy-atom tunnelling contributions as well as a large temperature-dependent H2O/D2O kinetic isotope effect of approximately 20, in excellent agreement with experiment.

Palaeo-environmental significance of evaporative calcite crusts in the Untersee Oasis, East Antarctica

Abstract Secondary carbonate precipitates on the surface of clasts have rarely been reported from Antarctica. Here, we infer the origin, age and palaeo-environmental significance of the calcite crusts in the Untersee Oasis, East Antarctica. The distribution of calcite crusts, which are up to 1 mm thick, is limited to locations with residual snow patches, and they have some of the highest δ18O (up to +17.4‰ Vienna Standard Mean Ocean Water (VSMOW)) and δ13C (up to +14.6‰ Vienna Pee Dee Belemnite (VPDB)) compositions of any carbonate deposits in terrestrial polar environments. Their δ18O and δ13C values are substantially enriched with respect to the isotopic values expected from equilibrium precipitation from the δ18O and δ13CDIC (DIC = dissolved inorganic carbon) of snow meltwater. The formation of the calcite crusts is ascribed to the evaporation of residual snow meltwater and the low relative humidity and strong winds, favouring a kinetic isotope effect. The 14C age distribution of the calcite crusts (1550 cal yr bp to modern) provides a minimum age for ice retreat and drainage of the palaeo-lake in Aurkjosen Cirque. However, in this polar desert environment in which surface melting is limited, the calcite crusts require sufficient snow accumulation and air temperatures warm enough to generate meltwater, and their age distribution corresponds to the late Holocene warm-wet climate period.

Enantioselective Si−H Insertion of Arylvinyldiazoesters Promoted by Rhodium(I)/Diene Complexes

AbstractAn enantioselective Si−H insertion between α‐arylvinyldiazoesters and silanes has been developed by employing rhodium(I)/chiral diene complex as the catalyst. This protocol offers a catalytic asymmetric approach to various chiral silanes containing α‐arylvinyl functionality at room temperature with good enantiocontrol (89–99% ee). Transformation of the resulting enantiomerically enriched α‐arylvinyl‐α‐silyl esters led to a variety of other valuable building blocks. The deuterium kinetic isotope effect experiments revealed a rhodium(I)‐mediated concerted Si−H insertion mechanism.

Accurate Quantum Dynamics Calculations for the Cl + CH4/CHD3/CD4 Reaction Rates.

Full-dimensional quantum dynamics simulations of the reaction of Cl with methane and its isotopomers are reported. Thermal rate constants are computed for the Cl + CH4 → HCl + CH3, Cl + CHD3 → HCl + CD3, and Cl + CD4 → DCl + CD3 reactions. Temperatures between 200 and 500 K are considered. In this temperature range, excellent agreement with the experiment is obtained. A detailed analysis of the kinetic isotope effect reveals the crucial importance of the CH3/CD3 umbrella motion. Comparison with approximate ring-polymer molecular dynamics simulations shows significant differences depending on the isotope studied.

Oxygenolytic cleavage of 1,2-diols with dioxygen by a mononuclear nonheme iron complex: Mimicking the reaction of myo-inositol oxygenase

A mononuclear iron(II) complex, [(TpPh2)FeII(OTf)(CH3CN)] (1) (TpPh2 = hydrotris(3,5-diphenylpyrazol-1-yl)borate, OTf = triflate) has been isolated and its efficiency toward the aliphatic CC bond cleavage reaction of 1,2-diols with dioxygen has been investigated. Separate reactions between 1 and different 1,2-diolates form the corresponding iron(II)-diolate complexes in solution. While the iron(II) complex of the tetradentate TPA (tris(2-pyridylmethyl)amine) ligand is not efficient in affecting the CC cleavage of 1,2-diol with dioxygen, complex 1 displays catalytic activity to afford carboxylic acid and aldehyde. Isotope labeling studies with 18O2 reveal that one oxygen atom from dioxygen is incorporated into the carboxylic acid product. The oxygenative CC cleavage reactions occur on the 1,2-diols containing at least one α-H atom. The kinetic isotope effect value of 5.7 supports the abstraction of an α-H by an iron(III)-superoxo species to propagate the CC cleavage reactions. The oxidative cleavage of 1,2-diolates by the iron(II) complex mimics the reaction catalyzed by the nonheme diiron enzyme, myo-inositol oxygenase.

CHIRAL SWITCHING CONTROL OF PHARMACEUTICAL SUBSTANCES

Objective: The aim of this study was to demonstrate that chiral switching should be recognized as a widespread phenomenon that extends beyond the production of pure enantiomeric drugs. Methods: To investigate the optical activity of substances from various chemical classes, enantiomers of chiral compounds (Sigma-Aldrich, USA) were chosen: valine and its racemic form (D-valine, L-valine, and racemic valine with optical purity ≥ 99%), L-ascorbic acid (content ≥ 99%), carbohydrates (D-glucose, D-galactose, L-galactose, contents ≥ 99.5%). Solutions were prepared using deuterium-depleted water (DDW–"light" water, D/H=4 ppm), natural deionized high-ohmic water (BD, D/H=140 ppm), and heavy water (99.9% D2O; Sigma-Aldrich). Optical activity was measured using the Atago POL-1/2 polarimeter. Results: One of the components in the racemic medication mixture can act as an inert agent, exhibit toxicity, or undergo undesirable biotransformation mechanisms, resulting in the formation of products with unknown properties. It has been established that a change in the deuterium/protium (D/H) ratio in water leads to a change in the equilibrium and kinetic characteristics of optically active compounds across various chemical classes, such as amino acids, carboxylic acids, and carbohydrates. An inequality was observed in the absolute values of the optical rotation of the L-and D-isomers of valine and galactose, depending on the D/H isotope ratio. The impact of chiral water clusters on optical rotation accounts for the sudden shift in the specific rotation of dilute solutions (less than 0.5%) of L-ascorbic acid in water, based on the D/H ratio. The influence of the isotopic composition of water was confirmed by studying the temperature-dependent mutarotation kinetics of D-glucose and L-and D-galactose in Arrhenius coordinates. The mutarotation process in natural high-resistivity water is characterized by an activation energy (Ea) of 40.8±1.4 kJ mol-1, while in deuterium-depleted water, Ea = 63.6±3.5 kJ mol-1. This results in a kinetic isotope effect for deuterium (KIED) of 1.6. Conclusion: Methodological approaches have been developed to control chiral switching based on the isotopic composition of water in vivo and in vitro. The study of changes in the optical activity of hierarchical structures in the human body, the influence of solvent properties on the mechanisms of optical rotation, as well as the use of KIED values, can be utilized to monitor various chiral transitions in vitro and living organisms.

Large enrichments in fatty acid 2H/1H ratios distinguish respiration from aerobic fermentation in yeast Saccharomyces cerevisiae

Shifts in the hydrogen stable isotopic composition (2H/1H ratio) of lipids relative to water (lipid/water 2H-fractionation) at natural abundances reflect different sources of the central cellular reductant, NADPH, in bacteria. Here, we demonstrate that lipid/water 2H-fractionation (2εfattyacid/water) can also constrain the relative importance of key NADPH pathways in eukaryotes. We used the metabolically flexible yeast Saccharomyces cerevisiae, a microbial model for respiratory and fermentative metabolism in industry and medicine, to investigate 2εfattyacid/water. In chemostats, fatty acids from glycerol-respiring cells were >550‰ 2H-enriched compared to those from cells aerobically fermenting sugars via overflow metabolism, a hallmark feature in cancer. Faster growth decreased 2H/1H ratios, particularly in glycerol-respiring cells by 200‰. Variations in the activities and kinetic isotope effects among NADP+-reducing enzymes indicate cytosolic NADPH supply as the primary control on 2εfattyacid/water. Contributions of cytosolic isocitrate dehydrogenase (cIDH) to NAPDH production drive large 2H-enrichments with substrate metabolism (cIDH is absent during fermentation but contributes up to 20 percent NAPDH during respiration) and slower growth on glycerol (11 percent more NADPH from cIDH). Shifts in NADPH demand associated with cellular lipid abundance explain smaller 2εfattyacid/water variations (<30‰) with growth rate during fermentation. Consistent with these results, tests of murine liver cells had 2H-enriched lipids from slower-growing, healthy respiring cells relative to fast-growing, fermenting hepatocellular carcinoma. Our findings point to the broad potential of lipid 2H/1H ratios as a passive natural tracker of eukaryotic metabolism with applications to distinguish health and disease, complementing studies that rely on complex isotope-tracer addition methods.

Cobalt(II)-Catalyzed Synthesis of γ-Diketones from Aryl Alkenes and Its Utilization in the Synthesis of Various Heterocyclic Compounds.

An earth-abundant Co(II) salt-catalyzed mild and affordable synthetic route has been developed for the synthesis of industrially relevant 1,4-dicarbonyl compounds (or γ-diketones) via oxidative coupling between aryl alkenes and ketones (both cyclic and acyclic) using TBHP and DBU as the oxidant and base, respectively. 1,4-Dicarbonyl compounds are known to be synthesized using expensive metal catalysts, dual catalysts, or low-cost metal complexes combined with an additive or ligand template, which further needs to be synthesized. Herein, we report the synthesis of 1,4-dicarbonyl compounds using cobalt(II) acetate as a catalyst without any expensive co-catalyst or ligand templates. This methodology has a broad substrate scope with significant yields and good functional group tolerance. Generation of unsymmetrical 1,4-dicarbonyls at room temperature and its versatile synthetic expansion to produce synthetically and biologically valuable heterocyclic compounds are salient features of this novel methodology. In addition, various controlled experiments such as primary kinetic isotope effect study, Hammett analysis with variation of the nature of the substituents on the styrene ring, and theoretical calculations (density functional theory) unravel the mechanistic intricacies involved in this new, simple, and atom-economic methodology.

Compound-specific, intra-molecular, and clumped 13C fractionations in the thermal generation and decomposition of ethane and propane: A DFT and kinetic investigation

The distribution of stable isotopes in organic compounds within geochemical systems reflects the reactions they undergo, which are often kinetically governed and can be elucidated through quantification based on first principles: employing quantum chemical simulations to derive energy barriers and kinetic isotope effects (KIEs), followed by kinetic calculations using these parameters. In this paper, the distribution of 13C in hydrocarbon gases during the thermal generation and decomposition of ethane and propane has been addressed. The work is based on abundant and systematic isotopic data, including the compound- and position-specific 13C compositions, recent improvements in kinetic networks in hydrocarbon generation, and state-of-the-art quantum chemical methods that can process organic geochemical systems involving solid phases. Density Functional Theory computations validate the carbonium path of hydrocarbon generation and the free-radical path of their decomposition under geological conditions. The first step of each path (alkyl sidechain protonation for generation and hydrogen abstraction for decomposition) is the rate-limiting step and determines the 13C fractionations of gaseous hydrocarbons, with the reaction rate constrained by the concentration or accessibility of the active species (hydrated protons and surface free radicals in the two paths, respectively). Alkyl protonation has normal 13C KIEs, but hydrogen abstraction on the surfaces of solid organic matter has inverse ones (substituting 12C by 13C accelerates the reaction). These inverse KIEs explain three isotopic phenomena in gaseous hydrocarbons in their late thermal evolution: 1) the depletion of 13C in ethane and propane accompanied by their decreased fraction in hydrocarbon gases (isotope “rollover”); 2) the reversed 13C sequence with carbon number (δ13Cmethane > δ13Cethane > δ13Cpropane); and 3) the depletion of 13C in the center carbon atoms of residual propane. Isotopic fractionation between gaseous hydrocarbons upon generation is evaluated using the Successive Random Chain-Scission model. The covariation of δ13C in alkanes shows a depletion in CH4 compared with the values expected from the reciprocal of the carbon numbers, indicating that this deviation may be intrinsic instead of mixing with microbial methane. The comparison of intramolecular 13C fractionation of propane between computational and observed data suggests the contribution of isoprenoids at the beginning of the generation, in addition to isotopic homogenization between the center and end carbon atoms through the ring closure of intermediate cations. The 13C–13C clumping exhibits a weak KIE in both the generation and decomposition of ethane; the clumping is essentially stochastically governed. This work demonstrates how first-principle calculations can provide significant insights into the details of geochemical reactions governing the isotopic distributions.

One-pot synthesis of multivariate porphyrinic metal-organic frameworks with the insertion of metallobipyridine and the cooperative catalysis for carbonyl hydrosilylation

A series of multivariate porphyrinic metal-organic frameworks with the insertion of bipyridine were obtained via one-pot synthesis. The single Pt site was introduced into the bipyridine linker of BPy@Co-PCN-224 through post-synthetic metalation. The resultant Pt(BPy)Cl2@M-PCN-224, especially when M was Co(II), could efficiently promote carbonyl hydrosilylation with a turnover number up to 13,140 based on Pt and could be recycled and reused for at least 6 times without a noticeable decrease of activity. Mechanistic studies based on in situ UV–vis, FT-IR and 1H NMR spectra, and kinetic isotope effect (KIE) experiments disclosed that the catalytic reaction was accomplished by the cooperative catalysis of Pt(BPy)Cl2 and Co-PCN-224, in which Pt(BPy)Cl2 was the catalytically active center and meanwhile the Co-PCN-224 could condense and increase the concentrations of ketone molecules in its inner cavities through the interactions between Co-porphyrin and carbonyl group. The plausible reaction pathway was proposed based on density functional theory calculations.

Dual C–Cl isotope fractionation offers potential to assess biodegradation of 1,2-dichloropropane and 1,2,3-trichloropropane by Dehalogenimonas cultures

1,2-dichloropropane (1,2-DCP) and 1,2,3-trichloropropane (1,2,3-TCP) are hazardous chemicals frequently detected in groundwater near agricultural zones due to their historical use in chlorinated fumigant formulations. In this study, we show that the organohalide-respiring bacterium Dehalogenimonas alkenigignens strain BRE15 M can grow during the dihaloelimination of 1,2-DCP and 1,2,3-TCP to propene and allyl chloride, respectively. Our work also provides the first application of dual isotope approach to investigate the anaerobic reductive dechlorination of 1,2-DCP and 1,2,3-TCP. Stable carbon and chlorine isotope fractionation values for 1,2-DCP (ƐC = −13.6 ± 1.4 ‰ and ƐCl = −27.4 ± 5.2 ‰) and 1,2,3-TCP (ƐC = −3.8 ± 0.6 ‰ and ƐCl = −0.8 ± 0.5 ‰) were obtained resulting in distinct dual isotope slopes (Λ12DCP = 0.5 ± 0.1, Λ123TCP = 4 ± 2). However direct comparison of ΛC-Cl among different substrates is not possible and investigation of the C and Cl apparent kinetic isotope effects lead to the hypothesis that concerted dichloroelimination mechanism is more likely for both compounds. In fact, whole cell activity assays using cells suspensions of the Dehalogenimonas-containing culture grown with 1,2-DCP and methyl viologen as electron donor suggest that the same set of reductive dehalogenases was involved in the transformation of 1,2-DCP and 1,2,3-TCP. This study opens the door to the application of isotope techniques for evaluating biodegradation of 1,2-DCP and 1,2,3-TCP, which often co-occur in groundwaters near agricultural fields.