Quartz Crystal Microbalance as a Holistic Detector for Quantifying Complex Organic Matrices during Liquid Chromatography: 2. Compound-Specific Isotope Analysis.

In carbon-compound-specific isotope analysis (carbon CSIA) of environmental micropollutants, purification of samples is often required to guarantee accurate measurements of a target compound. A companion paper has brought forward an innovative approach to couple a quartz crystal microbalance (QCM) with high-performance liquid chromatography (HPLC) for the online quantification of matrices during a gradient HPLC purification. This work investigates the benefit for isotope analysis of polar micropollutants typically present in environmental samples. Here, we studied the impact of the natural organic matter (NOM) on the isotopic integrity of model analytes and the suitability of the NOM-to-analyte ratio as a proxy for the sample purity. We further investigated limitations and enhancement of HPLC purification using QCM on C18 and C8 phases for single and multiple targets. Strong isotopic shifts of up to 3.3% toward the isotopic signature of NOM were observed for samples with an NOM-to-analyte ratio ≥10. Thanks to QCM, optimization of matrix removal of up to 99.8% of NOM was possible for late-eluting compounds. The efficiency of HPLC purification deteriorated when aiming for simultaneous purification of two or three compounds, leading to up to 2.5% less NOM removal. Our results suggest that one optimized HPLC purification can be achieved through systematic screening of 3 to 5 different gradients, thereby leading to a shift of the boundaries of accurate carbon CSIA by up to 2 orders of magnitude toward lower micropollutant concentrations.

Quartz Crystal Microbalance as a Holistic Detector for Quantifying Complex Organic Matrices during Liquid Chromatography: 1. Coupling, Characterization, and Validation.

A matrix in highly complex samples can cause adverse effects on the trace analysis of targeted organic compounds. A suitable separation of the target analyte(s) and matrix before the instrumental analysis is often a vital step for which chromatographic cleanup methods remain one of the most frequently used strategies, particularly high-performance liquid chromatography (HPLC). The lack of a simple real-time detection technique that can quantify the entirety of the matrix during this step, especially with gradient solvents, renders optimization of the cleanup challenging. This paper, along with a companion one, explores the possibilities and limitations of quartz crystal microbalance (QCM) dry-mass sensing for quantifying complex organic matrices during gradient HPLC. To this end, this work coupled a QCM and a microfluidic spray dryer with a commercial HPLC system using a flow splitter and developed a calibration and data processing strategy. The system was characterized in terms of detection and quantification limits, with LOD = 4.3-15 mg/L and LOQ = 16-52 mg/L, respectively, for different eluent compositions. Validation of natural organic matter in an environmental sample against offline total organic carbon analysis confirmed the approach's feasibility, with an absolute recovery of 103 ± 10%. Our findings suggest that QCM dry-mass sensing could serve as a valuable tool for analysts routinely employing HPLC cleanup methods, offering potential benefits across various analytical fields.

Selectivity of ß-Cyclodextrin Polymer toward Aquatic Contaminants: Insights from Ultrahigh-Resolution Mass Spectrometry of Dissolved Organic Matter.

Selectivity in solid-phase extraction (SPE) materials has become increasingly important for analyte enrichment in sensitive analytical workflows to alleviate detrimental matrix effects. Molecular-level investigation of matrix constituents, which are preferentially extracted or excluded, can provide the analytical chemist with valuable information to learn about their control on sorbent selectivity. In this work, we employ nontargeted Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) to elucidate the molecular chemodiversity of freshwater-derived dissolved organic matter (DOM) extracted by the selective model sorbent ß-cyclodextrin polymer (ß-CDP) in comparison to conventional, universal SPE sorbents (i.e., Oasis HLB, Supel-Select HLB, and LiChrolut EN). Statistical analysis of MS data corroborated the highly selective nature of ß-CDP by revealing the extracted DOM spectra that are most dissimilar to original compositions. We found that its selectivity was characterized by pronounced discrimination against highly oxygenated and unsaturated DOM compounds, which were associated with the classes of lignin-like, tannin-like, and carboxylic-rich alicyclic molecules. In contrast, conventional sorbents excluded less highly oxygenated compounds and showed a more universal extraction behavior for a wide range of DOM compositional space. We lay these findings in a larger context that aids the analyst in obtaining an a priori estimate of sorbent selectivity toward any target analyte of interest serving thereby an optimization of sample preparation. This study highlights the great value of nontargeted ultrahigh-resolution MS for better understanding of targeted analytics and provides new insights into the selective sorption behavior of novel sorbents.

Discriminative Behavior of Cyclodextrin Polymers against Dissolved Organic Matter: Role of Cavity Size and Sorbate Properties.

Cyclodextrin polymers (CDPs) are promising next-generation adsorbents in water purification technologies. The selectivity of the polymer derivate cross-linked with tetrafluoroterephthalonitrile (TFN-CDP) for nonionic and cationic micropollutants (MPs) over dissolved organic matter (DOM) renders the adsorbent also attractive for many analytical applications. The molecular drivers of the observed selectivity are, nonetheless, not yet fully understood. To provide new insights into the sorption mechanism, we (i) synthesized TFN-CDPs with different cavity sizes (α-, ß-, γ-CDP); (ii) assessed their extraction efficiencies for selected nonionic MPs in competition with different DOM size fractions (<1, 1-3, 3-10, >10 kDa) to test for size-selectivity; and (iii) performed nontargeted, ultrahigh resolution Fourier transform ion cyclotron resonance mass spectrometry analysis on CDP-extracted DOM compounds (<1 kDa) to probe for molecular sorbate properties governing their selective sorption. First, no evidence of size-selectivity was obtained through either the different CD cavity sizes (i) or the two independent approaches (ii) and (iii). Second, we found a dominant impact of sorbate oxygenation and polarity on the extraction of DOM and MPs, respectively, with relatively oxygen-poor/nonpolar molecules favorably retained on all α-, ß-, and γ-CDP. Third, our data indicates exclusion of an anionic matrix, such as carboxylic acids, but preferential sorption of cationic nitrogen-bearing DOM, pointing at repulsive and attractive forces with the negatively charged cross-linker as a likely reason. Therefore, we ascribe TFN-CDP's selectivity to nonpolar and electrostatic interactions between MPs/DOM and the polymer building blocks. These molecular insights can further aid in the optimization of efficient and selective sorbent design for environmental and analytical applications.

Avoiding Interferences in Advance: Cyclodextrin Polymers to Enhance Selectivity in Extraction of Organic Micropollutants for Carbon Isotope Analysis.

Compound-specific isotope analysis (CSIA) of organic water contaminants can provide important information about their sources and fate in the environment. Analyte enrichment from water remains nonetheless a critical yet inevitable step before measurement. Commercially available solid-phase extraction (SPE) sorbents are inherently nonselective leading to co-extraction of concurrent dissolved organic matter (DOM) and in turn to analytical interferences, especially for low-occurring contaminants. Here, we (i) increased extraction selectivity by synthesizing cyclodextrin polymers (α-, ß-, γ-CDP) as SPE sorbents, (ii) assessed their applicability to carbon isotope analysis for a selection of pesticides, and (iii) compared them with commonly used commercial sorbents. Extraction with ß-CDP significantly reduced backgrounds in gas chromatography-isotope ratio mass spectrometry (GC-IRMS) and enhanced sensitivity by a factor of 7.5, which was further confirmed by lower carbon-normalized CDOM/Canalyte ratios in corresponding extracts as derived from dissolved organic carbon (DOC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Gibbs free energies of adsorption demonstrated weak competition between DOM and analyte on the three CDPs. No isotopic fractionation (Δδ13C within ± 0.3‰) was observed for the investigated pesticides after using ß-CDP as an SPE sorbent covering a range of concentrations (5-500 µg L-1), flow velocities (5-40 cm min-1), and sorbent regeneration (up to six times). The present study highlights the benefit of selecting innovative extraction sorbents to avoid interferences in advance. This strategy in combination with existing cleanup approaches offers new prospects for CSIA at field concentrations of tens to hundreds of nanograms per liter.

Discovering Nature's Fingerprints: Isotope Ratio Analysis on Bioanalytical Mass Spectrometers.

For a generation or more, the mass spectrometry that developed at the frontier of molecular biology was worlds apart from isotope ratio mass spectrometry, a label-free approach done on optimized gas-source magnetic sector instruments. Recent studies show that electrospray-ionization Orbitraps and other mass spectrometers widely used in the life sciences can be fine-tuned for high-precision isotope ratio analysis. Since isotope patterns form everywhere in nature based on well-understood principles, intramolecular isotope measurements allow unique insights into a fascinating range of research topics. This Perspective introduces a wider readership to current topics in stable isotope research with the aim of discussing how soft-ionization mass spectrometry coupled with ultrahigh mass resolution can enable long-envisioned progress. We highlight novel prospects of observing isotopes in intact polar compounds and speculate on future directions of this adventure into the overlapping realms of biology, chemistry, and geology.

Microbial hydrolysis of atrazine in contaminated groundwater.

Degradation of the widespread herbicide atrazine has been intensively studied in soils, while its degradation in groundwater has received less attention. This work studied atrazine degradation in contaminated groundwater adjacent to its production plant. The degradation potential was first explored in groundwater enrichment cultures. A broad potential for microbial atrazine degradation was observed when atrazine served as the sole nitrogen source, even when incubated with nitrate. Hydroxyatrazine was formed by the cultures, while desethylatrazine and desisopropylatrazine were not detected. Both the atzA and the trzN genes were identified by quantitative PCR analysis, with a clear dominance of atzA. Carbon isotope enrichments throughout the degradation process varied between the different cultures, with ε values ranging from -0.6 to -5.5‰. This implies corresponding uncertainties when using compound-specific isotope analysis to estimate degradation extents. In the field samples, in-situ degradation was reflected by a high percentage of metabolites, with hydroxyatrazine accounting for >95% of the metabolites in most wells. Both atzA and trzN were detected in the groundwater at quantities of ≈102 to 106 copies mL-1, with a dominance of atzA over trzN. These results provide evidence of the high potential for atrazine hydrolysis in the contaminated groundwater.

Toward Improved Bioremediation Strategies: Response of BAM-Degradation Activity to Concentration and Flow Changes in an Inoculated Bench-Scale Sediment Tank.

Compound-specific isotope analysis (CSIA) can reveal mass-transfer limitations during biodegradation of organic pollutants by enabling the detection of masked isotope fractionation. Here, we applied CSIA to monitor the adaptive response of bacterial degradation in inoculated sediment to low contaminant concentrations over time. We characterized Aminobacter sp. MSH1 activity in a flow-through sediment tank in response to a transient supply of elevated 2,6-dichlorobenzamide (BAM) concentrations as a priming strategy and took advantage of an inadvertent intermittence to investigate the effect of short-term flow fluctuations. Priming and flow fluctuations yielded improved biodegradation performance and increased biodegradation capacity, as evaluated from bacterial activity and residual concentration time series. However, changes in isotope ratios in space and over time evidenced that mass transfer became increasingly limiting for degradation of BAM at low concentrations under such stimulated conditions, and that activity decreased further due to bacterial adaptation at low BAM (µg/L) levels. Isotope ratios, in conjunction with residual substrate concentrations, therefore helped identifying underlying limitations of biodegradation in such a stimulated system, offering important insight for future optimization of remediation schemes.

NO2 and natural organic matter affect both soot aggregation behavior and sorption of S-metolachlor.

Soot is an important carbonaceous nanoparticle (CNP) frequently found in natural environments. Its entry into surface waters can occur directly via surface runoff or infiltration, as well as via atmospheric deposition. Pristine soot is likely to rapidly undergo aggregation and subsequent sedimentation in aquatic environments. Further, soot can sorb a variety of organic contaminants, such as S-metolachlor (log KD = 3.25 ± 0.12). During atmospheric transport, soot can be chemically transformed by reactive oxygen species including NO2. The presence of natural organic matter (NOM) in surface waters can further affect the aquatic fate of soot. To better understand the processes driving the fate of soot and its interactions with contaminants, pristine and NO2-transformed model soot suspensions were investigated in the presence and absence of NOM. NO2-oxidized soot showed a smaller particle size, a higher number of particles remaining in suspension, and a decreased sorption of S-metolachlor (log KD = 2.47 ± 0.40). In agreement with findings for other CNPs, soot stability against aggregation was increased for both pristine and NO2 transformed soot in the presence of NOM.

A robust optimization technique for analysis of multi-tracer experiments.

Fate and transport of solutes in heterogeneous porous media is largely affected by diffusive mass exchange between mobile and immobile water zones. Since it is difficult to directly measure and determine the effect in the aquifers, multi-tracer experiments in combination with mathematical modeling are used to obtain quantitative information about unknown system parameters such as the effective mobile and immobile porosity, and the diffusive mass exchange between mobile and immobile water zones. The Single Fissure Dispersion Model (SFDM) describing nonreactive transport of solutes in saturated dual-porosity media, has been employed as a modeling approach to explain dual-porosity experiments in the field and laboratory (column experiments). SFDM optimization with conventional methods of minimization was immensely difficult due to its complex analytical form. Thus, previous studies used a trial and error procedure to fit it to the experimental observations. In this study, a rigorous optimization technique based on the newly developed scatter search method is presented that automatically minimizes the SFDM to find the optimal values of the hydrogeologically related parameters. The new program (OptSFDM) is accompanied with an easy-to-use graphical user interface (GUI) that is flexible and fully integrated. The program usability is showcased by a few, previously presented experimental case studies, and compared against the currently available, trial-and-error based, command-line executable, SFDM code.

Solid-phase extraction method for stable isotope analysis of pesticides from large volume environmental water samples.

Compound-specific isotope analysis (CSIA) is a valuable tool for assessing the fate of organic pollutants in the environment. However, the requirement of sufficient analyte mass for precise isotope ratio mass spectrometry combined with prevailing low environmental concentrations currently limits comprehensive applications to many micropollutants. Here, we evaluate the upscaling of solid-phase extraction (SPE) approaches for routine CSIA of herbicides. To cover a wide range of polarity, a SPE method with two sorbents (a hydrophobic hypercrosslinked sorbent and a hydrophilic sorbent) was developed. Extraction conditions, including the nature and volume of the elution solvent, the amount of sorbent and the solution pH, were optimized. Extractions of up to 10 L of agricultural drainage water (corresponding to up to 200 000-fold pre-concentration) were successfully performed for precise and sensitive carbon and nitrogen CSIA of the target herbicides atrazine, acetochlor, metolachlor and chloridazon, and metabolites desethylatrazine, desphenylchloridazon and 2,6-dichlorobenzamide in the sub-µg L-1-range. 13C/12C and 15N/14N ratios were measured by gas chromatography-isotope ratio mass spectrometry (GC/IRMS), except for desphenylchloridazon, for which liquid chromatography (LC/IRMS) and derivatization-GC/IRMS were used, respectively. The method validated in this study is an important step towards analyzing isotope ratios of pesticide mixtures in aquatic systems and holds great potential for multi-element CSIA applications to trace pesticide degradation in complex environments.

Molecularly Imprinted Polymers for Compound-Specific Isotope Analysis of Polar Organic Micropollutants in Aquatic Environments.

Compound-specific isotope analysis (CSIA) of polar organic micropollutants in environmental waters requires a processing of large sample volumes to obtain the required analyte masses for analysis by gas chromatography/isotope-ratio mass spectrometry (GC/IRMS). However, the accumulation of organic matter of unknown isotopic composition in standard enrichment procedures currently compromises the accurate determination of isotope ratios. We explored the use of molecularly imprinted polymers (MIPs) for selective analyte enrichment for 13C/12C and 15N/14N ratio measurements by GC/IRMS using 1 H-benzotriazole, a typical corrosion inhibitor in dishwashing detergents, as example of a widely detected polar organic micropollutant. We developed procedures for the treatment of >10 L of water samples, in which custom-made MIPs enabled the selective cleanup of enriched analytes in organic solvents obtained through conventional solid-phase extractions. Hydrogen bonding interactions between the triazole moiety of 1 H-benzotriazole, and the MIP were responsible for selective interactions through an assessment of interaction enthalpies and 15N isotope effects. The procedure was applied successfully without causing isotope fractionation to river water samples, as well as in- and effluents of wastewater treatment plants containing µg/L concentrations of 1 H-benzotriazole and dissolved organic carbon (DOC) loads of up to 28 mg C/L. MIP-based treatments offer new perspectives for CSIA of organic micropollutants through the reduction of the DOC-to-micropollutant ratios.

Simulation Studies to Explore Biodegradation in Water-Sediment Systems: From OECD 308 to OECD 309.

Studies according to OECD 308 and OECD 309 are performed to simulate the biodegradation of chemicals in water-sediment systems in support of persistence assessment and exposure modeling. However, several shortcomings of OECD 308 have been identified that hamper data evaluation and interpretation, and its relation to OECD 309 is still unclear. The present study systematically compares OECD 308 and OECD 309 and two variants thereof to derive recommendations on how to experimentally address any shortcomings and improve data for persistence and risk assessment. To this end, four (14)C-labeled compounds with different biodegradation and sorption behavior were tested across standard OECD 308 and 309 test systems and two modified versions thereof. The well-degradable compounds showed slow equilibration and the least mineralization in OECD 308, whereas the modified systems provided the highest degree of mineralization. Different lines of evidence suggest that this was due to increased oxygenation of the sediment in the modified systems. Particularly for rapidly degrading compounds, non-extractable residue formation was in line with degradation and did not follow the sediment-water ratio. For the two more slowly degrading compounds, sorption in OECD 309 (standard and modified) increased with time beyond levels proposed by equilibrium partitioning, which could be attributed to the grinding of the sediment through the stirring of the sediment suspension. Overall, the large differences in degradation observed across the four test systems suggest that refined specifications in test guidelines are required to reduce variability in test outcomes. At the same time, the amount of sediment and its degree of oxygenation emerged as drivers across all test systems. This suggests that a unified description of the systems was possible and would pave the way toward a more consistent consideration of degradation in the water-sediment systems across different exposure situations and regulatory frameworks.

Degradation of chlorotriazine pesticides by sulfate radicals and the influence of organic matter.

Atrazine, propazine, and terbuthylazine are chlorotriazine herbicides that have been frequently used in agriculture and thus are potential drinking water contaminants. Hydroxyl radicals produced by advanced oxidation processes can degrade these persistent compounds. These herbicides are also very reactive with sulfate radicals (2.2-3.5 × 10(9) M(-1) s(-1)). However, the dealkylated products of chlorotriazine pesticides are less reactive toward sulfate radicals (e.g., desethyl-desisopropyl-atrazine (DEDIA; 1.5 × 10(8) M(-1) s(-1))). The high reactivity of the herbicides is largely due to the ethyl or isopropyl group. For example, desisopropyl-atrazine (DIA) reacts quickly (k = 2 × 10(9) M(-1) s(-1)), whereas desethyl-atrazine (DEA) reacts more slowly (k = 9.6 × 10(8) M(-1) s(-1)). The tert-butyl group does not have a strong effect on reaction rate, as shown by the similar second order reaction rates between desethyl-terbuthylazine (DET; k = 3.6 × 10(8) M(-1) s(-1)) and DEDIA. Sulfate radicals degrade a significant proportion of atrazine (63%) via dealkylation, in which deethylation significantly dominates over deisopropylation (10:1). Sulfate and hydroxyl radicals react at an equally fast rate with atrazine (k (hydroxyl radical + atrazine) = 3 × 10(9) M(-1) s(-1)). However, sulfate and hydroxyl radicals differ considerably in their reaction rates with humic acids (k (sulfate radical + humic acids) = 6.8 × 10(3) L mgC(-1) s(-1) (mgC = mg carbon); k (hydroxyl radical + humic acids) = 1.4 × 10(4) L mgC(-1) s(-1)). Thus, in the presence of humic acids, atrazine is degraded more efficiently by sulfate radicals than by hydroxyl radicals.

Formation of bromate in sulfate radical based oxidation: mechanistic aspects and suppression by dissolved organic matter.

Sulfate radical based oxidation is discussed being a potential alternative to hydroxyl radical based oxidation for pollutant control in water treatment. However, formation of undesired by-products, has hardly been addressed in the current literature, which is an issue in other oxidative processes such as bromate formation in ozonation of bromide containing water (US-EPA and EU drinking water standard of bromate: 10 µg L(-1)). Sulfate radicals react fast with bromide (k = 3.5 × 10(9) M(-1) s(-1)) which could also yield bromate as final product. The mechanism of bromate formation in aqueous solution in presence of sulfate radicals has been investigated in the present paper. Further experiments were performed in presence of humic acids and in surface water for investigating the relevance of bromate formation in context of pollutant control. The formation of bromate by sulfate radicals resembles the well described mechanism of the hydroxyl radical based bromate formation. In both cases hypobromous acid is a requisite intermediate. In presence of organic matter formation of bromate is effectively suppressed. That can be explained by formation of superoxide formed in the reaction of sulfate radicals plus aromatic moieties of organic matter, since superoxide reduces hypobromous acid yielding bromine atoms and bromide. Hence formation of bromate can be neglected in sulfate radical based oxidation at typical conditions of water treatment.