Method validation and environmental monitoring of triethanolamine ester quaternary ammonium compounds.

In this study water and sediment samples, collected from the River Nene (Northamptonshire) at several sites in the vicinity of the Great Billing sewage treatment plant (STP), were analysed for triethanolamine quaternary compounds (TEAQ, ester quats). A method was developed using liquid chromatography tandem mass spectrometry (LC/MS/MS) with a electrospray ionisation source (ESI). Ten components were determined using a characterised commercial sample of Tallow TEAQ as a standard. To our knowledge this is the first time environmental concentrations of a wide spectrum of individual homologues of TEAQ have been reliably quantified covering a broad range of environmental matrices (STP influent, STP effluent, surface waters and sediments), due to the challenging nature of the analytical method. The method featured novel solutions for the determination of long and multiple chain length alkyl quats, controlling loss processes, background contamination and chromatographic performance. TEAQ compounds were found to be highly removed in the sewage treatment plant resulting in low effluent concentrations. Low concentrations in both river water and sediment samples were found also. In many cases levels were below the Method Detection Limit (MDL). In river water samples, mean values of TEAQ compounds found were 210-398 ng/L for C16:0/C18:0 TEAQ diester and 126-287 ng/L for C18:0/C18:0 TEAQ diester. River sediment was found to contain mean TEAQ levels of 7.07-12.5, 19.7 to 40.3 and 7.04-35.1 µg/kg dry weight for C16:0/C16:0, C16:0/C18:0, and C18:0/C18:0 TEAQ, respectively. At Great Billing STP monoesters and diesters of TEAQ were shown to be efficiently removed (>97 and 99 %, respectively), although limited samples were taken on this occasion.

Are Non-animal Systemic Safety Assessments Protective? A Toolbox and Workflow.

An important question in toxicological risk assessment is whether non-animal new approach methodologies (NAMs) can be used to make safety decisions that are protective of human health, without being overly conservative. In this work, we propose a core NAM toolbox and workflow for conducting systemic safety assessments for adult consumers. We also present an approach for evaluating how protective and useful the toolbox and workflow are by benchmarking against historical safety decisions. The toolbox includes physiologically based kinetic (PBK) models to estimate systemic Cmax levels in humans, and 3 bioactivity platforms, comprising high-throughput transcriptomics, a cell stress panel, and in vitro pharmacological profiling, from which points of departure are estimated. A Bayesian model was developed to quantify the uncertainty in the Cmax estimates depending on how the PBK models were parameterized. The feasibility of the evaluation approach was tested using 24 exposure scenarios from 10 chemicals, some of which would be considered high risk from a consumer goods perspective (eg, drugs that are systemically bioactive) and some low risk (eg, existing food or cosmetic ingredients). Using novel protectiveness and utility metrics, it was shown that up to 69% (9/13) of the low risk scenarios could be identified as such using the toolbox, whilst being protective against all (5/5) the high-risk ones. The results demonstrated how robust safety decisions could be made without using animal data. This work will enable a full evaluation to assess how protective and useful the toolbox and workflow are across a broader range of chemical-exposure scenarios.

High-Throughput Assessment of the Abiotic Stability of Test Chemicals in In Vitro Bioassays.

Abiotic stability of chemicals is not routinely tested prior to performing in vitro bioassays, although abiotic degradation can reduce the concentration of test chemicals leading to the formation of active or inactive transformation products, which may lead to misinterpretation of bioassay results. A high-throughput workflow was developed to measure the abiotic stability of 22 test chemicals in protein-rich aqueous media under typical bioassay conditions at 37 °C for 48 h. These test chemicals were degradable in the environment according to a literature review. The chemicals were extracted from the exposure media at different time points using a novel 96-pin solid-phase microextraction. The conditions were varied to differentiate between various reaction mechanisms. For most hydrolyzable chemicals, pH-dependent degradation in phosphate-buffered saline indicated that acid-catalyzed hydrolysis was less important than reactions with hydroxide ions. Reactions with proteins were mainly responsible for the depletion of the test chemicals in the media, which was simulated by bovine serum albumin (BSA) and glutathione (GSH). 1,2-Benzisothiazol-3(2H)-one, 2-methyl-4-isothiazolinone, and l-sulforaphane reacted almost instantaneously with GSH but not with BSA, indicating that GSH is a good proxy for reactivity with electrophilic amino acids but may overestimate the actual reaction with three-dimensional proteins. Chemicals such as hydroquinones or polyunsaturated chemicals are prone to autoxidation, but this reaction is difficult to differentiate from hydrolysis and could not be simulated by the oxidant N-bromosuccinimide. Photodegradation played a minor role because cells are exposed in incubators in the dark and simulations with high light intensities did not yield realistic degradation. Stability predictions from various in silico prediction models for environmental conditions can give initial indications of the stability but were not always consistent with the experimental stability in bioassays. As the presented workflow can be performed in high throughput under realistic bioassay conditions, it can be used to provide an experimental database for developing bioassay-specific stability prediction models.

Fate of household and personal care chemicals in typical urban wastewater treatment plants indicate different seasonal patterns and removal mechanisms.

Studies on the presence and fate of household and personal care chemicals (HPCCs) in wastewater treatment plants (WWTPs) are important due to their increasing consumption worldwide. The seasonal patterns and removal mechanisms of HPCCs are not well understood for WWTPs that apply different treatment technologies. To answer these questions, the sewage and sludge samples were taken from 10 typical WWTPs in Northeast China. Levels of UV filters in the influents in the warm season were significantly greater than that of the cold season (p < 0.05). Significant seasonal differences were found for the removals of many HPCCs. Results revealed that the highest removal efficiencies were found for linear alkylbenzene sulphonates with values ranging from 97.2% to 99.7%, and the values were 50.0%-99.9% for other HPCCs. The SimpleTreat model demonstrated that the studied WWTPs were operating with high efficiency at the time of sampling. The sorption of HPCCs to sludge can be strongly associated with their physicochemical parameters. Mass balance calculation suggested that sorption was the dominant mechanism for the removal of antimicrobials, while degradation and/or biotransformation were the other mechanisms for removing the most HPCCs in the WWTPs. This study real the factors influencing the seasonal patterns and removal mechanisms which imply the need for further studies to fully understands the plant and human health implications as sludge could be used in the municipal land application of biosolids.

Biodegradation Kinetics of Fragrances, Plasticizers, UV Filters, and PAHs in a Mixture─Changing Test Concentrations over 5 Orders of Magnitude.

Biodegradation of organic chemicals emitted to the environment is carried out by mixed microbial communities growing on multiple natural and xenobiotic substrates at low concentrations. This study aims to (1) perform simulation type biodegradation tests at a wide range of mixture concentrations, (2) determine the concentration effect on the biodegradation kinetics of individual chemicals, and (3) link the mixture concentration and degradation to microbial community dynamics. Two hundred ninety-four parallel test systems were prepared using wastewater treatment plant effluent as inoculum and passive dosing to add a mixture of 19 chemicals at 6 initial concentration levels (ng/L to mg/L). After 1-30 days of incubation at 12 °C, abiotic and biotic test systems were analyzed using arrow solid phase microextraction and GC-MS/MS. Biodegradation kinetics at the highest test concentrations were delayed for several test substances but enhanced for the reference chemical naphthalene. Test concentration thus shifted the order in which chemicals were degraded. 16S rRNA gene amplicon sequencing indicated that the highest test concentration (17 mg C/L added) supported the growth of the genera Acidovorax, Novosphingobium, and Hydrogenophaga, whereas no such effect was observed at lower concentrations. The chemical and microbial results confirm that too high mixture concentrations should be avoided when aiming at determining environmentally relevant biodegradation data.

Comparison of passive-dosed and solvent spiked exposures of pro-carcinogen, benzo[a]pyrene, to human lymphoblastoid cell line, MCL-5.

Genotoxicity testing methods in vitro provide a means to predict the DNA damaging effects of chemicals on human cells. This is hindered in the case of hydrophobic test compounds, however, which will partition to in vitro components such as plastic-ware and medium proteins, in preference to the aqueous phase of the exposure medium. This affects the freely available test chemical concentration, and as this freely dissolved aqueous concentration is that bioavailable to cells, it is important to define and maintain this exposure. Passive dosing promises to have an advantage over traditional 'solvent spiking' exposure methods and involves the establishment and maintenance of known chemical concentrations in the in vitro medium, and therefore aqueous phase. Passive dosing was applied in a novel format to expose the MCL-5 human lymphoblastoid cell line to the pro-carcinogen, benzo[a]pyrene (B[a]P) and was compared to solvent (dimethyl sulphoxide) spiked B[a]P exposures over 48 h. Passive dosing induced greater changes, at lower concentrations, to micronucleus frequency, p21 mRNA expression, cell cycle abnormalities, and cell and nuclear morphology. This was attributed to a maintained, definable, free chemical concentration using passive dosing and the presence or absence of solvent, and highlights the influence of exposure choice on genotoxic outcomes.

Validation of sampling techniques and SPE-UPLC/MS/MS for home and personal care chemicals in the Songhua Catchment, Northeast China.

A method for the simultaneous determination of 18 home and personal care chemicals (HPCCs) in river water and wastewater was developed using solid-phase extraction and ultra-high-performance liquid chromatography with tandem mass spectrometry (SPE and UPLC/MS/MS). A series of tests were designed to find the potential background interference and loss of HPCCs during the sample preservation and pretreatment process. Our results suggested that a considerable amount of some target compounds were lost with increasing days of storage even at 4 °C. Accordingly, the samples were stored acidified and refrigerated, including during transportation, to reduce the losses. Experiments on filtration of water samples suggested that recoveries of many HPCCs were significantly affected by the filtration. It is therefore recommended to avoid filtration of water samples where possible. The internal standard corrected recoveries for the HPCCs ranged from 64.2 to 107.0%, except for benzisothiazolone which did not have an appropriate internal standard, in river water which was considered to be the most difficult matrix. The method detection limits for river water, influent and effluent samples were in the ranges of 0.17 to 42 ng/L, 13 to 5100 ng/L, and 0.50 to 200 ng/L, respectively. The validated method was applied for the determination of HPCCs in sewage water collected from a full-scale wastewater treatment plant (WWTP) in a typical urban city in Northeast China and from river water upstream and downstream of the WWTP. Linear alkylbenzene sulphonate, caffeine, methyl paraben, benzalkonium chloride, triclocarban, and triclosan were the major compounds detected in the river water and wastewater samples. Sampling variability for the WWTP (intra-day and inter-day) and cross-river was also determined with the purpose of designing future monitoring requirements. Small variations in these samples confirmed that composite samples and a single sampling event would be representative for future use.

A flow-through aqueous standard generation system for thin film microextraction investigations of UV filters and biocides partitioning to different environmental compartments.

In this paper problems associated with preparation of aqueous standard of highly hydrophobic compounds such as partial precipitation, being lost on the surfaces, low solubility in water and limited sample volume for accurate determination of their distribution coefficients are addressed. The following work presents two approaches that utilize blade thin film microextraction (TFME) to investigate partitioning of UV filters and biocides to humic acid (dissolved organic carbon) and sediment. A steady-state concentration of target analytes in water was generated using a flow-through aqueous standard generation (ASG) system. Dialysis membranes, a polytetrafluoroethylene permeation tube, and a frit porous (0.5 µm) coated by epoxy glue were basic elements used for preparation of the ASG system. In the currently presented study, negligible depletion TFME using hydrophilic-lipophilic balance (HLB) and octadecyl silica-based (C18) sorbents was employed towards the attainment of free concentration values of target analytes in the studied matrices. Thin film geometry provided a large volume of extraction phase, which improved the sensitivity of the method towards highly matrix-bound analytes. Extractions were performed in the equilibrium regime so as to prevent matrix effects and with aims to reach maximum method sensitivity for all analytes under study. Partitioning of analytes on dissolved organic carbon (DOC) was investigated in ASG to facilitate large sample volume conditions. Binding percentages and DOC distribution coefficients (Log KDOC) ranged from 20 to 98% and 3.71-6.72, respectively. Furthermore, sediment-water partition coefficients (Kd), organic-carbon normalized partition coefficients (Log KOC), and DOC distribution coefficients (Log KDOC) were investigated in slurry sediment, and ranged from 33 to 2860, 3.31-5.24 and 4.52-5.75 Lkg-1, respectively. The obtained results demonstrated that investigations utilizing ASG and TFME can yield reliable binding information for compounds with high log KOW values. This information is useful for study of fate, transport, and ecotoxicological effects of UV filters and biocides in aquatic environment.

Time Weighted Average Concentration Monitoring Based on Thin Film Solid Phase Microextraction.

Time weighted average (TWA) passive sampling with thin film solid phase microextraction (TF-SPME) and liquid chromatography tandem mass spectrometry (LC-MS/MS) was used for collection, identification, and quantification of benzophenone-1, benzophenone-2, benzophenone-3, benzophenone-4, 2-phenylbenzimidazole-5-sulfonic acid, octocrylene, octylmethoxycinnamate, butylmethoxydibenzoylmethane, triclocarban and triclosan in the aquatic environment. Two types of TF-SPME passive samplers, including a retracted thin film device using a hydrophilic lipophilic balance (HLB) coating, and an open bed configuration with an octadecyl silica-based (C18)coating, were evaluated in an aqueous standard generation system. Laboratory calibration results indicated that the thin film retracted device using HLB coating is suitable to determine TWA concentrations of polar analytes in water, with an uptake that was linear up to 70 days. In open bed form, a one-calibrant kinetic calibration technique was accomplished by loading benzophenone3-d5 as calibrant on the C18 coating to quantify all nonpolar compounds. The experimental results showed that the one-calibrant kinetic calibration technique can be used for determination of classes of compounds in cases where deuterated counterparts are either not available or expensive. The developed passive samplers were deployed in wastewater-dominated reaches of the Grand River (Kitchener, ON) to verify their feasibility for determination of TWA concentrations in on-site applications. Field trials results indicated that these devices are suitable for long-term and short-term monitoring of compounds varying in polarity, such as UV blockers and biocide compounds in water, and the data were in good agreement with literature data.

Measurement of cyclic volatile methylsiloxanes in the aquatic environment using low-density polyethylene passive sampling devices using an in-field calibration study--challenges and guidance.

Cyclic volatile methylsiloxanes (cVMS) are used in personal care products and are hydrophobic, volatile and persistent. Their environmental water concentrations are low and are difficult to detect using conventional sampling methods. This study shows the potential of passive sampling for cVMS. We used low-density polyethylene (LDPE) samplers and in-field calibration methods for octamethylcyclotetrasiloxane (D4) and decamethylcyclopentasiloxane (D5). (13)C-D4 and (13)C-D5, methyltris(trimethylsiloxy)silane (MT), tetrakis(trimethylsiloxy)silane (TK), and five deuterated polycyclic aromatic hydrocarbons (PAHs) were used as performance reference compounds (PRCs). Samplers were calibrated (7-d) using effluent at a treatment plant, with uptake of cVMS and losses of the PRCs measured at 12 time-points. Concentrations of D4 (53ngL(-1)) and D5 (1838ngL(-1)) were stable in the effluent. Uptake of D4 and loss of (13)C-D4 were isotropic and equilibrium was approached by 7-d. Two estimates of sampler uptake rate (Rs) were 2.1Ld(-1) and 2.5Ld(-1). The estimated log LDPE/water partition coefficient was 4.4. The uptake of D5 was slower (Rs=0.32Ld(-1)) and equilibrium was not reached. Offloading of (13)C-D5, MT and TK were slow, and isotropic behaviour was not demonstrated for D5. Offloading of PAHs followed the predicted pattern for LDPE. Uptake of cVMS appeared to be under membrane control, due to low diffusion coefficients in LDPE. Samplers can monitor time-weighted average concentrations of D4 for less than a week, and D5 for longer periods. LDPE samplers allow cVMS to be determined at lower concentrations than by spot sampling methods.

Thin-film microextraction coupled to LC-ESI-MS/MS for determination of quaternary ammonium compounds in water samples.

The dual nature of the quaternary ammonium compounds, having permanently charged hydrophilic quaternary ammonium heads and long-chain hydrophobic tails, makes the sample preparation step and analysis of these compounds challenging. A high-throughput method based on thin-film solid-phase microextraction (SPME) and liquid chromatography mass spectrometry was developed for simultaneous quantitative analysis of nine benzylic and aliphatic quaternary ammonium compounds. Chromatographic separation and detection of analytes were obtained in reverse-phase mode in 8 min using a triple quadrupole mass spectrometer. Hydrophilic lipophilic balance particle-coated blades were found to be the most suitable among the different coatings tested in terms of recoveries and carryover on the blades. For desorption solvents, 70/30, v/v (A/B) with 0.1 % formic acid (where A is 10 mM ammonium acetate in acetonitrile/water (95/5 , v/v) and B is 0.1 % (v/v) formic acid in isopropyl alcohol) was shown to be the most efficient solvent for the desorption of the analytes from the SPME sorbent. The SPME method was optimised in terms of extraction, pH, and preconditioning, as well as extraction and desorption times. Optimum conditions were 45 min of extraction time and 15 min of desorption time, all with agitation. The extraction was found to be optimum in a range of pH 6.0 to 8.0, which is consistent with the natural pH of water samples. Wide linear dynamic ranges with the developed method were obtained for each compound, enabling the application of the method for a wide range of concentrations. The developed method was validated according to the Food and Drug Administration criteria. The proposed method is the first SPME-based approach describing the applicability of the high-throughput thin-film SPME in a 96-well system for analysis of such challenging compounds.

Simulated use and wash-off release of decamethylcyclopentasiloxane used in anti-perspirants.

The cyclic volatile methylsiloxane, decamethylcyclopentasiloxane (D5) is used in a large variety of personal care products. Based on the physical-chemical properties of D5, it is likely that losses due to volatilisation may strongly influence the levels entering the aquatic environment. The aim of this study was to quantify the amount of D5 in waste wash water, after typical application and use in a range of deodorant and anti-perspirant (AP) products. Results implied significant losses after a 24h period (>99.9%), and suggest that the use of D5 in leave-on products, such as deodorants/AP is not likely to contribute a significant down-the-drain emission source. An illustrative example is presented, based on data reporting the use of D5 in a range of personal care products (both wash-off and leave-on), which suggests that the contribution of D5 used in wash-off products to the aquatic environment may be considerably more significant. Limitations associated with our understanding of the actual D5 inclusion levels in the products, the market share of the products containing D5, and the variability of consumer habits, are identified as data gaps that need to be addressed in order to better refine down-the-drain emission estimates.

Monitoring and modelling of siloxanes in a sewage treatment plant in the UK.

Monitoring of cyclic volatile methylsiloxanes (cVMS) carried out at Anglian Water's Broadholme sewage treatment plant (STP) is described. The method deployed used headspace gas chromatography/mass spectrometry (HS-GC/MS) and the addition of isotopically labelled cVMS to correct for partitioning in samples containing high levels of particulate and dissolved organic carbon. The method was capable of measuring cVMS in raw sewage samples, with recoveries of 80%, 85% and 71% respectively, for D4, D5 and D6. The limit of quantification was 0.2µgL(-1) for all three substances. Recoveries close to 100% were observed for all cVMS spiked into treated effluent (LOQ=0.01µgL(-1)). Despite the volatile nature of cVMS and its ubiquitous presence in the ambient atmosphere, the methods deployed showed excellent recoveries, reproducibility and quantification limits. A distinct diurnal variation in cVMS concentration, probably linked with the use of personal care products was observed for raw sewage but not in treated sewage effluent. The estimated per capita consumption of D5 (∼2.7mgcap(-1)d(-1)) derived for the population served by this plant was significantly lower than that derived in the Environment Agency (UK) risk assessment (11.6mgcap(-1)d(-1)). The cVMS were highly removed during sewage treatment with efficiencies greater than 98%. The methods and findings of this pilot study can be used as the basis for future studies on the fate of cVMS substances in STPs.

Determination of decamethylcyclopentasiloxane in river and estuarine sediments in the UK.

Robust analytical procedures for the measurement of decamethylcyclopentasiloxane (D5) in river and estuarine sediments and their application in determining environmental concentrations in the UK are presented for the first time in this work. Novel approaches to minimise commonly reported artefacts are utilised, improving the confidence in the concentrations of D5 reported. Accelerated solvent extraction (ASE) and liquid-solid extraction methods are compared. Both methods use on-column injection gas chromatography/mass spectrometry (GC/MS). Measurements of D5 concentrations in sediments sampled from the river Great Ouse and from the Humber estuary (UK) are also reported. ASE was suitable to measure concentrations of D5 in sediments obtained from the river Great Ouse, UK (186-1450 ng g⁻¹, dry weight) and octamethyltetracyclosiloxane (D4, 12-24 ng g⁻¹, dry weight). C12 linear alkybenzene (C12 LAB), which can be used as a chemical marker for sewage effluent related emissions, was also measured in this analysis. Liquid-solid extraction was optimised to provide more confidence in the lower D5 concentrations measured in the Humber estuary, UK (49-256 ng g⁻¹, dry weight). A Limit of quantitation (LOQ) for D5 of 57-110 and 4 ng g⁻¹ dry weight was determined for ASE and liquid-solid extraction, respectively.

Determination of decamethylcyclopentasiloxane in river water and final effluent by headspace gas chromatography/mass spectrometry.

A method is described for the analysis of decamethylcyclopentasiloxane (D(5)) in river water and treated waste water using headspace gas chromatography/mass spectrometry. Internal standard addition to samples and field blanks was carried out in the field to provide both a measure of recovery and to prevent any exposure of samples to laboratory air, which contained background levels of D(5). Measured levels of D(5) were typically in the range <10-29ngL(-1) in the River Great Ouse (UK) with slightly higher levels in the River Nene (UK). The measured concentration of D(5) in treated waste water varied between 31 and 400ngL(-1), depending on the type of treatment process employed.

Biodegradability of highly ethoxylated nonionic surfactants: determination of intermediates and pathways of biodegradation.

Existing data regarding alcohol ethoxylate (AE) surfactants indicate that structures with greater than 20 ethoxylate (EO) units per molecule or with multibranched alkyl chains may not pass a ready biodegradability test. This could have important consequences for complying with regional regulatory requirements and for the potential risks these chemicals could present to the environment. We investigated the influence of chemical structure on the biodegradability of AEs with different alkyl chain branching and EO content. The AEs investigated were a multibranched AE (average, 18 EO), an oxo-AE (monobranched; average, 23 EO), and a linear AE of oleochemical origin (average, 40 EO). The aims of the present study were to assess the ready biodegradability of AEs with high EO content and to establish the mechanism or pathway by which biodegradation occurs for the oxo-AE. Biodegradation studies were conducted using standard test conditions (International Standards Organization 14593). Solid-phase extraction and liquid chromatography with electrospray mass spectrometry were used to detect profiles in both derivatized and underivatized extracts of samples. Derivatization with phthalic anhydride was used to improve ionization of the lower-ethoxylated AEs and free alcohol that were key indicators in the present study. All AEs were rapidly biodegraded, achieving more than 60% mineralization of parent material. Central fission was the predominant mechanism for the oxo-AE, as confirmed by identification of the oligomeric distribution and quantification of polyethylene glycol released during biodegradation.