Tracking the transformation of persistent organic pollutants in food webs using multi element isotope and enantiomer fractionation.

In order to track the transformation of persistent organic pollutants (POPs) in food webs, field experiments were conducted at two sites using stable isotope and enantiomer fractionation concepts. The enantiomers of α-hexachlorocyclohexane (α-HCH) were selected as representative compounds for POPs. Isotope and enantiomer fractionation allowed the characterization of α-HCH enantiomer biotransformation processes along trophic levels of the food web - from soil and plants to animal livers, fat tissues and milk. The enrichment of heavy isotopes in soils, plants and sediments as well as the changes of enantiomer fractionation indicate that the biotransformation of α-HCH occurred in these compartments. Moreover, the increase of carbon and chlorine isotopic compositions as well as the changes of enantiomer fractionation of liver, fat tissues and milk demonstrated that the overall HCH exposure was much higher than estimates based on concentration levels, while the isotope and enantiomer fractionation revealed the enantiomer specific enantiomer uptake across the blood-brain barriers. Dual element isotope analysis suggested that complex transformation processes have occurred along the potential food web from the HCH sources over different environmental compartments to animal livers, fat tissues and milk. The results imply that the analyses of stable isotope compositions and concentrations has potential to reconstruct the exposure of higher organisms to POPs.

Determination of Stable Hydrogen Isotopic Composition and Isotope Enrichment Factor at Low Hydrogen Concentration.

Determination of stable hydrogen isotopic compositions (δ2H) is currently challenged to achieve a high detection limit for reaching the linear range where δ2H values are independent of concentration. Therefore, it is difficult to assess precise δ2H values for calculating the hydrogen isotope enrichment factor (εH) and for field application where the concentrations of contaminants are relatively low. In this study, a data treatment approach was developed to obtain accurate δ2H values below the linear range. The core concept was to use a logarithmic function to fit the δ2H values below the linear range and then adjust the δ2H values below the linear range into the linear range by using the fitted logarithmic equation. Moreover, the adjusted δ2H values were calibrated by using laboratory reference materials, e.g., n-alkanes. Tris(2-chloroethyl) phosphate (TCEP) and hexachlorocyclohexane (HCH) isomers were selected as examples of complex heteroatom-bearing compounds to develop the data treatment approach. This data treatment approach was then tested using δ2H values from a TCEP transformation experiment with OH radicals. Comparable δ2H values and εH between the low-concentration experiment and the reference experiment were obtained using the developed approach. Therefore, the developed data treatment approach enables a possibility of determining the hydrogen isotopic compositions of organic components in low concentrations. It is especially valuable for determining organic contaminants in environmental samples, which are usually present in low concentrations.

Compound-Specific Carbon, Hydrogen, and Nitrogen Isotope Analysis of Nitro- and Amino-Substituted Chlorobenzenes in Complex Aqueous Matrices.

Compound-specific isotope analysis (CSIA) is an established tool to study the fate of legacy groundwater contaminants but is only emerging for nonconventional contaminants, e.g., nitro- and amino-substituted chlorobenzenes that are widely used as industrial feedstock and the target of this work. To date, CSIA of the target compound groups used special combustion interfaces and the potential matrix interferences in environmental samples has not been assessed. We validated CSIA methods for δ13C, δ2H, and δ15N of four analytes from each chemical group and developed a solid-phase extraction (SPE) method to minimize matrix interferences during preconcentration of complex aqueous samples. The SPE recovery was >80% and the method quantification limits of SPE-CSIA for δ13C, δ2H, and δ15N were 0.03-0.57, 1.3-2.7, and 3.4-10.2 µM aqueous-phase concentrations, respectively, using 2 L of spiked MQ water. The SPE-CSIA procedure showed negligible isotope fractionation for δ13C (≤0.5‰), δ15N (≤0.5‰), and δ2H (≤5‰ for nitroaromatics and ≤10‰ for aminoaromatics). In addition, solvent evaporation, water sample storage up to 7 months, and SPE extract storage for 1.5 years did not change analytes' δ13C signatures beyond ±0.5‰. However, to avoid significant δ2H and δ15N fractionation of aminoaromatics, cartridge breakthrough should be avoided and SPE preconcentration must be conducted at pH > pKa + 2. Application of the method at a contaminated site showed excellent precision, at ≤0.3‰ for C and N, and ≤1.5‰ for H. The methods validated here now allow the use of multielement CSIA to track the environmental fate of nitro- and amino-substituted chlorobenzenes in complex aqueous samples.

Characterization of Hexachlorocyclohexane Isomer Dehydrochlorination by LinA1 and LinA2 Using Multi-element Compound-Specific Stable Isotope Analysis.

Dehydrochlorination is one of the main (thus far discovered) processes for aerobic microbial transformation of hexachlorocyclohexane (HCH) which is mainly catalyzed by LinA enzymes. In order to gain a better understanding of the reaction mechanisms, multi-element compound-specific stable isotope analysis was applied for evaluating α- and γ-HCH transformations catalyzed by LinA1 and LinA2 enzymes. The isotopic fractionation (εE) values for particular elements of (+)α-HCH (εC = -10.8 ± 1.0‰, εCl = -4.2 ± 0.5‰, εH = -154 ± 16‰) were distinct from the values for (-)α-HCH (εC = -4.1 ± 0.7‰, εCl = -1.6 ± 0.2‰, εH = -68 ± 10‰), whereas the dual-isotope fractionation patterns were almost identical for both enantiomers (ΛC-Cl = 2.4 ± 0.4 and 2.5 ± 0.2, ΛH-C = 12.9 ± 2.4 and 14.9 ± 1.1). The εE of γ-HCH transformation by LinA1 and LinA2 were -7.8 ± 1.0‰ and -7.5 ± 0.8‰ (εC), -2.7 ± 0.3‰ and -2.5 ± 0.4‰ (εCl), -170 ± 25‰ and -150 ± 13‰ (εH), respectively. Similar ΛC-Cl values (2.7 ± 0.2 and 2.9 ± 0.2) were observed as well as similar ΛH-C values (20.1 ± 2.0 and 18.4 ± 1.9), indicating a similar reaction mechanism by both enzymes during γ-HCH transformation. This is the first data set on 3D isotope fractionation of α- and γ-HCH enzymatic dehydrochlorination, which gave a more precise characterization of the bond cleavages, highlighting the potential of multi-element compound-specific stable isotope analysis to characterize different transformation processes (e.g., dehydrochlorination and reductive dehalogenation).

Stormwater Bioretention Cells Are Not an Effective Treatment for Persistent and Mobile Organic Compounds (PMOCs).

Bioretention cells are a stormwater management technology intended to reduce the quantity of water entering receiving bodies. They are also used to reduce contaminant releases, but their performance is unclear for hydrophilic persistent and mobile organic compounds (PMOCs). We developed a novel eight-compartment one-dimensional (1D) multimedia model of a bioretention cell ("Bioretention Blues") and applied it to a spike and recovery experiment conducted on a system near Toronto, Canada, involving PMOC benzotriazole and four organophosphate esters (OPEs). Compounds with (log DOC) (organic carbon-water distribution coefficients) < ∼2.7 advected through the system, resulting in infiltration or underdrain flow. Compounds with log DOC > 3.8 were mostly sorbed to the soil, where subsequent fate depended on transformation. For compounds with 2.7 ≤ log DOC ≤ 3.8, sorption was sensitive to event size and compound-specific diffusion parameters, with more sorption expected for smaller rain events and for compounds with larger diffusion coefficients. Volatilization losses were minimal for all compounds tested. Direct uptake by vegetation also played a negligible role regardless of the compounds' physicochemical properties. Nonetheless, model simulations showed that vegetation could play a role by increasing transpiration, thereby increasing sorption to the bioretention soil and reducing PMOC release. Model results suggest design modifications to bioretention cells.

Stable isotope fractionation associated with the synthesis of hexachlorocyclohexane isomers for characterizing sources.

The stable isotope fingerprints of hexachlorocyclohexane (HCH) isomers have potential for identifying sources as they are related to the synthesis processes and isotopic compositions of raw materials. However, the isotopic fractionation associated with the synthesis processes has not been investigated. Therefore, photochemical synthesis experiments using benzene and chlorine gas were conducted to characterize the associated isotopic fractionation under different conditions. Different patterns of isotopic fractionation factors (αC, αCl, and αH) were observed in each experiment. The large variability of αH is related to the accumulating secondary hydrogen isotope effects or the rearrangement of C-H bonds at the cyclohexane ring. An increase of δ13C and δ37Cl values of HCH isomers was observed during synthesis, which is related to the C-Cl bond formation in the radical dichlorination forming HCH and the subsequent chlorine substitution forming heptachlorocyclohexanes. The large variability of δ2H values is related to the secondary and primary hydrogen isotope effects. Different δ13C, δ37Cl and δ2H values among HCH isomers were observed, indicating that conformational complexity of HCH caused by arrangement of C-Cl bonds in planar and axial positions also influence the isotope values. The understanding of isotopic fractionation during HCH synthesis can be indicative for source identification in the field.

Carbon, hydrogen and nitrogen stable isotope fractionation allow characterizing the reaction mechanisms of 1H-benzotriazole aqueous phototransformation.

1H-benzotriazole is part of a larger family of benzotriazoles, which are widely used as lubricants, polymer stabilizers, corrosion inhibitors, and anti-icing fluid components. It is frequently detected in urban runoff, wastewater, and receiving aquatic environments. 1H-benzotriazole is typically resistant to biodegradation and hydrolysis, but can be transformed via direct photolysis and photoinduced mechanisms. In this study, the phototransformation mechanisms of 1H-benzotriazole were characterized using multi-element compound-specific isotope analysis (CSIA). The kinetics, transformation products, and isotope fractionation results altogether revealed that 1H-benzotriazole direct photolysis and indirect photolysis induced by OH radicals involved two alternative pathways. In indirect photolysis, aromatic hydroxylation dominated and was associated with small carbon (εC = -0.65 ± 0.03‰), moderate hydrogen (εH = -21.6‰), and negligible nitrogen isotope enrichment factors and led to hydroxylated forms of benzotriazole. In direct photolysis of 1H-benzotriazole, significant nitrogen (εN = -8.4 ± 0.4 to -4.2 ± 0.3‰) and carbon (εC = -4.3 ± 0.2 to -1.64 ± 0.04‰) isotope enrichment factors indicated an initial N-N bond cleavage followed by nitrogen elimination with a C-N bond cleavage. The results of this study highlight the potential for multi-element CSIA application to track 1H-benzotriazole degradation in aquatic environments.

Characterizing the biotransformation of hexachlorocyclohexanes in wheat using compound-specific stable isotope analysis and enantiomer fraction analysis.

Hexachlorocyclohexane isomers (HCHs) are persistent organic pollutants being responsible for environmental contamination worldwide. In order to characterize transformation of HCHs in different plant compartments during uptake, a hydroponic experimental setup was designed using wheat as the test plant. The extent of transformation was determined by using compound-specific isotope analysis (CSIA) and enantiomer fraction (EF) analysis. In nutrient solutions, no change of carbon (δ13C) and chlorine isotope ratios (δ37Cl) of α-HCH and ß-HCH was detected throughout the experiment indicating no transformation there. In wheat leaves, stems and roots, however, transformation of α-HCH due to a C‒Cl bond cleavage was indicated by increasing δ13C and δ37Cl compared to the nutrient solution. In addition, 1,3,4,5,6-pentachlorocyclohexene (PCCH) was identified as the major metabolite of α-HCH transformation. For ß-HCH, in contrast, no transformation was detected. The evaluation of enantiomer fraction analysis revealed no change of the EF(-) in the nutrient solution or on root surface but a decrease in the wheat compartments, providing an evidence for the preferential biological transformation of (-)α-HCH in wheat. The current study provides the first experimental evidence for biotransformation of α-HCH in wheat using CSIA and EF and provides a concept to evaluate processes during phytoremediation.

Compound-Specific Isotope Analysis and Enantiomer Fractionation to Characterize the Transformation of Hexachlorocyclohexane Isomers in a Soil-Wheat Pot System.

The uptake by plants from soil is one of the first steps for hexachlorocyclohexane (HCH) isomers to enter the food web. However, the HCH transformation associated with the uptake process is still not well understood. Therefore, a soil-wheat pot experiment was conducted to characterize the HCH transformation during wheat growth using compound-specific isotope analysis (CSIA) and enantiomer fractionation. The results showed that the δ13C and δ37Cl values of ß-HCH remained stable in soil and wheat, revealing no transformation. In contrast, an increase of δ13C and δ37Cl values of α-HCH indicated its transformation in soil and wheat. A shift of the enantiomer fraction (EF) (-) from 0.50 to 0.35 in soil at the jointing stage and 0.35 to 0.57 at the harvest stage suggested that the preferential transformation of enantiomers varied at different growth stages. Based on the dual element isotope analysis, the transformation mechanism in the soil-wheat system was different from that in wheat in hydroponic systems. The high abundance of HCH degraders, Sphingomonas sp. and Novosphingobium sp., was detected in the α-HCH-treated rhizosphere soil, supporting the potential for biotransformation. The application of CSIA and EF allows characterizing the transformation of organic pollutants such as HCHs in the complex soil-plant systems.

Can Alkaline Hydrolysis of γ-HCH Serve as a Model Reaction to Study Its Aerobic Enzymatic Dehydrochlorination by LinA?

Hexachlorocyclohexane (HCH) isomers constitute a group of persistent organic pollutants. Their mass production and treatment have led to a global environmental problem that continues to this day. The characterization of modes of degradation of HCH by isotope fractionation is a current challenge. Multi isotope fractionation analysis provides a concept to characterize the nature of enzymatic and chemical transformation reactions. The understanding of the kinetic isotope effects (KIE) on bond cleavage reaction contributes to analyses of the mechanism of chemical and enzymatic reactions. Herein, carbon, chlorine, and hydrogen kinetic isotope effects are measured and predicted for the dehydrochlorination reaction of γ-HCH promoted by the hydroxyl ion in aqueous solution. Quantum mechanical (QM) microsolvation with an implicit solvation model and path integral formalism in combination with free-energy perturbation and umbrella sampling (PI-FEP/UM) and quantum mechanical/molecular mechanical QM/MM potentials for including solvent effects as well as calculating isotope effects are used and analyzed with respect to their performance in reproducing measured values. Reaction characterization is discussed based on the magnitudes of obtained isotope effects. The comparative analysis between the chemical dehydrochlorination of γ-HCH in aqueous media and catalyzed reaction by dehydrochlorinase, LinA is presented and discussed. Based on the values of isotope effects, these two processes seem to occur via the same net mechanism.

Isotope fractionation approach to characterize the reactive transport processes governing the fate of hexachlorocyclohexanes at a contaminated site in India.

The transformation processes of hexachlorocyclohexane isomers (HCHs) from production sites of Lindane across the landscape and along the food web were studied as an example to understand the fate of POPs in the environment. Therefore, we studied the concentration and isotope composition of HCHs in different matrices in the vicinity of a dumpsite and a chemical plant producing HCHs in India. Carbon isotope compositions (δ13C) of HCHs and the enantiomer fraction (EF) of α-HCH were used as indicators to characterize in situ degradation in soil, groundwater, and sediment as well as along the food web. The HCHs were detected in plants growing on contaminated soil. Elevated concentrations of HCHs were found in a number of crops, which indicates an important transfer pathway of HCHs entering food webs. The EF value of α-HCH and the δ13C signature of HCHs indicated that degradation processes occurred in the rhizosphere or within the plants potentially attenuating the contamination of HCHs. The isotope enrichment of HCHs in dung and milk samples showed that degradation of HCHs may take place in the digestive track of cow and buffalo as well as during their metabolism. The δ13C of HCHs was used to analyze the potential dispersion routes on the landscape scale in order to understand the reactive transport pathways starting at the source of HCHs. In this study, the potential of carbon isotope fractionation and EF for characterizing uptake of HCHs into plants and accumulation in the food web were examined. To the best of our knowledge, this is the first study using the combination of stable isotope fractionation and EF to track the reactive transport processes in a complex environment including the food web.

2H and 13C isotope fractionation analysis of organophosphorus compounds for characterizing transformation reactions in biogas slurry: Potential for anaerobic treatment of contaminated biomass.

The ability of anaerobic digestion (AD) to eliminate organophosphorus model compounds (OPs) with structural elements of phosphate, phosphorothioate and phosphorodithioate esters was studied. The enzymatic mechanism of the first irreversible degradation reaction was characterized using metabolite pattern and kinetic 2H/13C-isotope effect in original, cell-free and heat sterilized biogas slurry. The isotope fractionation study suggests different modes of degradation reactions. Representatives for phosphate ester, tris(2-chloroethyl) phosphate and tris(1,3-dichloro-2-propyl) phosphate, were hydrolyzed in biogas slurry without carbon or hydrogen isotope fractionation. Representatives for phosphorodithioate, Dimethoate and Malathion, were degraded in original slurry yielding carbon enrichment factor (εC) of -0.6 ±â€¯0.1‰ and -5.5 ±â€¯0.1‰ (-0.9 ±â€¯0.1‰ and -7.2 ±â€¯0.5‰ in cell-free slurry), without hydrogen isotope fractionation. Phosphorothioate degradation represented by Parathion and Parathion-methyl yielded surprisingly different εC (-0.7 ±â€¯0.2 and -3.6 ±â€¯0.4‰) and εH (-33 ±â€¯5 and -5 ±â€¯1‰) in original slurry compared to cell-free slurry (εC = -2.5 ±â€¯0.5 and -8.6 ±â€¯1.4‰; εH = -61 ±â€¯10 and -10 ±â€¯3‰) suggesting H-C bond cleavage. Degradation of Parathion and Parathion-methyl in sterilized slurry gave carbon but not hydrogen fractionation implying relative thermostable enzymatic activity with different mechanism. The correlation of 2H and 13C stable isotope fractionation of Parathion in biogas slurry showed distinct pattern (Λoriginal = 31 ±â€¯11, Λcell-free = 20 ±â€¯2), indicating different mechanism from chemical hydrolysis. Overall, AD can be a potential treatment for OPs contaminated biomass or contaminated organic waste material.

Enantiomer and Carbon Isotope Fractionation of α-Hexachlorocyclohexane by Sphingobium indicum Strain B90A and the Corresponding Enzymes.

Chiral organic contaminants, like α-hexachlorocyclohexane (α-HCH), showed isotope fractionation and enantiomer fractionation during biodegradation. This study aims to understand the correlation between these two processes. Initial tests of α-HCH degradation by six Sphingobium strains (with different LinA variants) were conducted. Results showed variable enantiomer selectivity over the time course. In contrast, constant enantiomer selectivity was observed in experiments employing (i) cell suspensions, (ii) crude extracts, or (iii) LinA1 and LinA2 enzymes of strain B90A for α-HCH degradation in enzyme activity assay buffer. The average value of enantioselectivity (ES) were -0.45 ± 0.03 (cell suspensions), -0.60 ± 0.05 (crude extracts), and 1 (LinA1) or -1 (LinA2). The average carbon isotope enrichment factors (εc) of (+)α- and (-)α-HCH were increased from cells suspensions (-6.3 ± 0.1‰ and -2.3 ± 0.03‰) over crude extracts (-7.7 ± 0.4‰ and -3.4 ± 0.02‰) to purified enzymes (-11.1 ± 0.3‰ and -3.8 ± 0.2‰). The variability of ES and the εc were discussed based on the effect of mass transport and degradation rates. Our study demonstrates that enantiomer and isotope fractionation of α-HCH are two independent processes and both are affected by underlying reactions of individual enzymes and mass transport to a different extent.

A concept for studying the transformation reaction of hexachlorocyclohexanes in food webs using multi-element compound-specific isotope analysis.

An analytical concept using stable isotope fractionation for analyzing persistent organic pollutants (POPs) in food webs was developed and tested. We have evaluated methods for the extraction and clean-up of hexachlorocyclohexane isomers (HCHs) as the model compounds of POPs from water, soil, plant, milk, fish oil and pork liver in order to study the reactive transport processes of HCHs in food webs using multi-element compound-specific isotope analysis (CSIA). The extraction and clean-up methods were evaluated for recovery efficiency and isotope effects. The precision and accuracy for carbon, hydrogen and chlorine isotope analysis were within the analytical precision of ±0.5‰, ±5‰ and ±0.3‰, respectively. The method was applied for stable isotope analysis of HCHs in possible food webs from soil to plants, and to animals. Isotope compositions of HCHs in cow/buffalo milk and dung, wild animal livers and seal blubber were obtained and compared to the sources of HCHs. The magnitude of isotope enrichment demonstrated the potential of CSIA for analyzing reactive transport processes of HCHs in the food web. The concept using multi-element stable isotope analysis can be applied for source identification, characterization of degradation mechanisms, and particularly contaminant accumulation in the food web, which demonstrates the potential in new scientific areas for CSIA.

Carbon and hydrogen isotopic fractionation during abiotic hydrolysis and aerobic biodegradation of phthalate esters.

We systematically investigated the changes of carbon and hydrogen isotope signatures of three phthalate esters (PAEs) during (i) abiotic hydrolysis over the pH range of 2, 7 and 10, and (ii) aerobic biodegradation initiated by hydrolysis by Rhodococcus opacus strain DSM 43250. Significant carbon isotopic fractionation was exhibited under all investigated conditions. Hydrogen isotopic fractionation was observed in some experiments and is hypothesized to be a secondary isotope effect due to the absence of a hydrogen bond cleavage during hydrolysis. Dual stable isotope analysis (Λ = Δδ2H/Δδ13C) resulting from abiotic hydrolysis and aerobic biodegradation showed similar magnitudes for dimethyl phthalate (DMP) and diethyl phthalate (DEP). The calculated carbon apparent kinetic isotope effects (AKIEC) for the hydrolytic pathway (CO bond cleavage) of PAEs fall within an expected range of 1.03-1.09, with the exception of lower AKIEC values for dibutyl phthalate (DBP) during hydrolysis at pH 2 and aerobic biodegradation. The lower AKIEC of DBP at pH 2 and aerobic biodegradation is likely related to a transition state from reactant-like to tetrahedral intermediate-like structure. Abiotic and biotic hydrolysis of PAEs resulted in similar AKIEC and Λ values due to the CO bond cleavage pathway, indicating the potential of dual isotope analysis to detect and quantify hydrolytic processes of PAEs in the environment. The pronounced primary carbon and typically low secondary or absent hydrogen isotopic fractionation might form a typical pattern to identify the hydrolytic reaction of PAEs in the environment. The characteristic Λ values of the hydrolytic reaction were different from Λ values of chemical oxidation of PAEs and showed diagnostic potential of dual HC isotope analysis to analyze reactions.

Carbon and hydrogen stable isotope analysis for characterizing the chemical degradation of tributyl phosphate.

Tributyl phosphate (TBP) belongs to the group of trialkyl substituted organophosphate esters. Its chemical reactivity depends on the stability of various chemical bonds. TBP was used as a model compound for the development of a concept using stable isotope fractionation associated with bond cleavage reactions for better understanding the fate of TBP in the environment. Carbon isotope enrichment factors (εC) of TBP hydrolysis were found to be pH dependent (-3.8 ±â€¯0.3‰ at pH 2, -4.6 ±â€¯0.5‰ at pH 7, -2.8 ±â€¯0.1‰ at pH 9, no isotope fractionation at pH 12), which is in accordance with the mode of a SN2 hydrolytic bond cleavage. Hydrogen isotope fractionation was negligible as no H bond cleavage is involved during hydrolysis. The apparent carbon kinetic isotope effect (AKIEC) ranged from 1.045 to 1.058. In contrast to hydrolysis, both carbon and hydrogen isotope fractionation were observed during radical oxidation of TBP by OH and SO4-, yielding εC from -0.9 ±â€¯0.1‰ to -0.5 ±â€¯0.1‰ and εH from -20 ±â€¯2‰ to -11 ±â€¯1‰. AKIEC and AKIEH varied from 1.007 to 1.011 and from 1.594 to 2.174, respectively. The correlation of 2H and 13C isotope fractionation revealed Λ values ranging from 17 ±â€¯1 to 25 ±â€¯6. Results demonstrated that the correlation of 2H and 13C isotope fractionation of TBP allowed to identify radical reactions and to distinguish them from hydrolysis. The presented dual isotope analysis approach has diagnostic value for characterizing the chemical transformation of TBP in the environment.

Carbon and hydrogen isotope analysis of parathion for characterizing its natural attenuation by hydrolysis at a contaminated site.

The applicability of compound-specific isotope analysis (CSIA) for assessing in situ hydrolysis of parathion was investigated in a contaminated aquifer at a former pesticide wastes landfill site. Stable isotope analysis of parathion extracted from groundwater taken from different monitoring wells revealed a maximum enrichment in carbon isotope ratio of +4.9‰ compared to the source of parathion, providing evidence that in situ hydrolysis took place. Calculations based on the Rayleigh-equation approach indicated that the natural attenuation of parathion was up to 8.6% by hydrolysis under neutral and acidic conditions. In degradation experiments with aerobic and anaerobic parathion-degrading microbes, no carbon and hydrogen isotope fractionation of parathion were observed. For the first time, CSIA has been applied for the exclusive assessment of the hydrolysis of phosphorothioate-containing organophosphorus pesticides at a contaminated field site.

Aqueous photodegradation of substituted chlorobenzenes: Kinetics, carbon isotope fractionation, and reaction mechanisms.

Substituted chlorobenzenes are the basic substructure of many surface water contaminants. In this study, the isotope fractionation and reaction mechanisms involved during the aqueous direct and indirect photodegradation of CH3-, Cl-, and NO2- substituted chlorobenzenes were investigated in laboratory experiments. Only 4-nitrochlorobenzene showed slow but isotopically fractionating direct photolysis. During indirect photodegradation using UV/H2O2-generated OH radicals, the pseudo first-order reaction rate constants increased in the order of the NO2- < Cl- < CH3- substituted chlorobenzenes. The most pronounced carbon enrichment factors were observed for nitrochlorobenzenes (up to -4.8 ±â€¯0.5‰), whereas the lowest were for chlorotoluenes (≤-1.0 ±â€¯0.1‰). As the substituents became more electron-withdrawing, the activation energy barrier increased, leading to slower reaction rates, and the transition state changed to a more symmetrical or less reactant-like structure, resulting in larger apparent kinetic isotope effects. The results suggest that the rate-determining step in the reaction with OH radicals was the addition of the electrophile to the benzene ring. Even though further research is needed to quantify isotope fractionation during other transformation processes, these results showed evidence that compound specific isotope analysis can be used as a diagnostic tool for the fate of substituted chlorobenzenes in water.

Characterizing chemical transformation of organophosphorus compounds by 13C and 2H stable isotope analysis.

Continuous and excessive use of organophosphorus compounds (OPs) has led to environmental contaminations which raise public concerns. This study investigates the isotope fractionation patterns of OPs in the aquatic environment dependence upon hydrolysis, photolysis and radical oxidation processes. The hydrolysis of parathion (EP) and methyl parathion (MP) resulted in significant carbon fractionation at lower pH (pH2-7, εC=-6.9~-6.0‰ for EP, -10.5~-9.9‰ for MP) but no detectable carbon fractionation at higher pH (pH12). Hydrogen fractionation was not observed during any of the hydrolysis experiments. These results indicate that compound specific isotope analysis (CSIA) allows distinction of two different pH-dependent pathways of hydrolysis. Carbon and hydrogen isotope fractionation were determined during UV/H2O2 photolysis of EP and tris(2-chloroethyl) phosphate (TCEP). The constant δ2H values determined during the OH radical reaction of EP suggested that the rate-limiting step proceeded through oxidative attack by OH radical on the PS bond. The significant H isotope enrichment suggested that OH radical oxidation of TCEP was caused by an H-abstraction during the UV/H2O2 processes (εH=-56±3‰). Fenton reaction was conducted to validate the H isotope enrichment of TCEP associated with radical oxidation, which yielded εH of -34±5‰. Transformation products of OPs during photodegradation were identified using Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR MS). This study highlights that the carbon and hydrogen fractionation patterns have the potential to elucidate the transformation of OPs in the environment.

Validation of GC-IRMS techniques for δ13C and δ2H CSIA of organophosphorus compounds and their potential for studying the mode of hydrolysis in the environment.

Compound-specific stable isotope analysis (CSIA) is among the most promising tools for studying the fate of organic pollutants in the environment. However, the feasibility of multidimensional CSIA was limited by the availability of a robust method for precise isotope analysis of heteroatom-bearing organic compounds. We developed a method for δ 13C and δ 2H analysis of eight organophosphorus compounds (OPs) with different chemical properties. In particular, we aimed to compare high-temperature conversion (HTC) and chromium-based HTC (Cr/HTC) units to explore the limitations of hydrogen isotope analysis of heteroatom-bearing compounds. Analysis of the amount dependency of the isotope values (linearity analysis) of OPs indicated that the formation of HCl was a significant isotope fractionation process leading to inaccurate δ 2H analysis in HTC. In the case of nonchlorinated OPs, by-product formation of HCN, H2S, or PH3 in HTC was observed but did not affect the dynamic range of reproducible isotope values above the limit of detection. No hydrogen-containing by-products were found in the Cr/HTC process by use of ion trap mass spectrometry analysis. The accuracy of gas chromatography - isotope ratio mass spectrometry was validated in comparison with elemental analyzer - isotope ratio mass spectrometry. Dual-isotope fractionation yielded Λ values of 0 ± 0 at pH 7, 7 ± 1 at pH 9, and 30 ± 6 at pH 12, indicating the potential of 2D CSIA to characterize the hydrolysis mechanisms of OPs. This is the first report on the combination of δ 2H and δ 13C isotope analysis of OPs, and this is the first study providing a systematic evaluation of HTC and Cr/HTC for hydrogen isotope analysis using OPs as target compounds. Graphical Abstract Comparison of δ2H measurement of non-chlorinated and chlorinated OPs via GC-Cr/HTC-IRMS and GC-HTC-IRMS system.