Non-target screening to trace ozonation transformation products in a wastewater treatment train including different post-treatments.

Ozonation and subsequent post-treatments are increasingly implemented in wastewater treatment plants (WWTPs) for enhanced micropollutant abatement. While this technology is effective, micropollutant oxidation leads to the formation of ozonation transformation products (OTPs). Target and suspect screening provide information about known parent compounds and known OTPs, but for a more comprehensive picture, non-target screening is needed. Here, sampling was conducted at a full-scale WWTP to investigate OTP formation at four ozone doses (2, 3, 4, and 5 mg/L, ranging from 0.3 to 1.0 gO3/gDOC) and subsequent changes during five post-treatment steps (i.e., sand filter, fixed bed bioreactor, moving bed bioreactor, and two granular activated carbon (GAC) filters, relatively fresh and pre-loaded). Samples were measured with online solid-phase extraction coupled to liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS) using electrospray ionization (ESI) in positive and negative modes. Existing non-target screening workflows were adapted to (1) examine the formation of potential OTPs at four ozone doses and (2) compare the removal of OTPs among five post-treatments. In (1), data processing included principal component analysis (PCA) and chemical knowledge on possible oxidation reactions to prioritize non-target features likely to be OTPs. Between 394 and 1328 unique potential OTPs were detected in positive ESI for the four ozone doses tested; between 12 and 324 unique potential OTPs were detected in negative ESI. At a specific ozone dose of 0.5 gO3/gDOC, 27 parent compounds were identified and were related to 69 non-target features selected as potential OTPs. Two OTPs were confirmed with reference standards (venlafaxine N-oxide and chlorothiazide); 34 other potential OTPs were in agreement with literature data and/or reaction mechanisms. In (2), hierarchical cluster analysis (HCA) was applied on profiles detected in positive ESI mode across the WWTP and revealed 11 relevant trends. OTP removal was compared among the five post-treatments and 54-83% of the non-target features that appeared after ozonation were removed, with the two GAC filters performing the best. Overall, these data analysis strategies for non-target screening provide a useful tool to understand the behavior of unknown features during ozonation and post-treatment and to prioritize certain non-targets for further identification.

Ozonation of Para-Substituted Phenolic Compounds Yields p-Benzoquinones, Other Cyclic α,ß-Unsaturated Ketones, and Substituted Catechols.

Phenolic moieties are common functional groups in organic micropollutants and in dissolved organic matter, and are exposed to ozone during drinking water and wastewater ozonation. Although unsubstituted phenol is known to yield potentially genotoxic p-benzoquinone during ozonation, little is known about the effects of substitution of the phenol ring on transformation product formation. With batch experiments employing differing ozone/target compound ratios, it is shown that para-substituted phenols ( p-alkyl, p-halo, p-cyano, p-methoxy, p-formyl, p-carboxy) yield p-benzoquinones, p-substituted catechols, and 4-hydroxy-4-alkyl-cyclohexadien-1-ones as common ozonation products. Only in a few cases did para-substitution prevent the formation of these potentially harmful products. Quantum chemical calculations showed that different reaction mechanisms lead to p-benzoquinone, and that cyclohexadienone can be expected to form if no such pathway is possible. These products can thus be expected from most phenolic moieties. Kinetic considerations showed that substitution of the phenolic ring results in rather small changes of the apparent second order rate constants for phenol-ozone reactions at pH 7. Thus, in mixtures, most phenolic structures can be expected to react with ozone. However, redox cross-reactions between different transformation products, as well as hydrolysis, can be expected to further alter product distributions under realistic treatment scenarios.

Evaluation of a full-scale wastewater treatment plant upgraded with ozonation and biological post-treatments: Abatement of micropollutants, formation of transformation products and oxidation by-products.

To protect the ecosystem and drinking water resources in Switzerland and in the countries of the downstream catchments, a new Swiss water protection act entered into force in 2016 aiming to reduce the discharge of micropollutants from wastewater treatment plants (WWTPs). As a consequence, selected WWTPs must be upgraded by an advanced treatment for micropollutant abatement with suitable and economic options such as (powdered) activated carbon treatment or ozonation. WWTP Neugut (105'000 people equivalent) was the first WWTP in Switzerland to implement a long-term full-scale ozonation. Differing specific ozone doses in the range of 0.35-0.97 g O3/g DOC were applied to determine the adequate ozone dose to fulfill the requirements of the Swiss water protection act. Based on this assessment, a specific ozone dose of 0.55 g O3/g DOC is recommended at this plant to ensure an average abatement of the twelve selected indicator substances by ≥80% over the whole treatment. A monitoring of 550 substances confirmed that this dose was very efficient to abate a broad range of micropollutants by >79% on average. After ozonation, an additional biological post-treatment is required to eliminate possible negative ecotoxicological effects generated during ozonation caused by biodegradable ozonation transformation products (OTPs) and oxidation by-products (OBPs). Three biological treatments (sand filtration, moving bed, fixed bed) and granular activated carbon (GAC, fresh and pre-loaded) filtration were evaluated as post-treatments after ozonation. In parallel, a fresh GAC filter directly connected to the effluent of the secondary clarifier was assessed. Among the three purely biological post-treatments, the sand filtration performed best in terms of removal of dissolved organic carbon (DOC), assimilable organic carbon (AOC) and total suspended solids (TSS). The fresh activated carbon filtration ensured a significant additional micropollutants abatement after ozonation due to sorption. The relative abatement of the indicator substances ranged between 20 and 89% after 27'000 bed volumes (BV) and was still substantial at 50'000 BV. In an identical GAC filter running in parallel and being fed with the effluent of the secondary clarifier, the elimination was less efficient. Seven primary OTPs (chlorothiazide and six N-oxides) formed during ozonation could be quantified thanks to available reference standards. Their concentration decreased with increasing specific ozone doses with the concomitant formation of other OTPs. The seven OTPs were found to be stable compounds and were not abated in the biological post-treatments. They were sorbed in the fresh GAC filter, but less efficiently than the corresponding parent compounds. Two OBPs, bromate (BrO3-) and N-nitrosodimethylamine (NDMA), were formed during ozonation but did not exceeded 5 µg/L for bromate and 30 ng/L for NDMA at the recommended specific ozone dose of 0.55 g O3/g DOC. NDMA was well abated in all post-treatments (minimum 41% during fixed bed filtration, maximum 83% during fresh GAC filtration), while bromate was very stable as expected.

Effect of operational and water quality parameters on conventional ozonation and the advanced oxidation process O3/H2O2: Kinetics of micropollutant abatement, transformation product and bromate formation in a surface water.

The efficiency of ozone-based processes under various conditions was studied for the treatment of a surface water (Lake Zürich water, Switzerland) spiked with 19 micropollutants (pharmaceuticals, pesticides, industrial chemical, X-ray contrast medium, sweetener) each at 1 µg L-1. Two pilot-scale ozonation reactors (4-5 m3 h-1), a 4-chamber reactor and a tubular reactor, were investigated by either conventional ozonation and/or the advanced oxidation process (AOP) O3/H2O2. The effects of selected operational parameters, such as ozone dose (0.5-3 mg L-1) and H2O2 dose (O3:H2O2 = 1:3-3:1 (mass ratio)), and selected water quality parameters, such as pH (6.5-8.5) and initial bromide concentration (15-200 µg L-1), on micropollutant abatement and bromate formation were investigated. Under the studied conditions, compounds with high second-order rate constants kO3>104 M-1 s-1 for their reaction with ozone were well abated (>90%) even for the lowest ozone dose of 0.5 mg L-1. Conversely, the abatement efficiency of sucralose, which only reacts with hydroxyl radicals (OH), varied between 19 and 90%. Generally, the abatement efficiency increased with higher ozone doses and higher pH and lower bromide concentrations. H2O2 addition accelerated the ozone conversion to OH, which enables a faster abatement of ozone-resistant micropollutants. Interestingly, the abatement of micropollutants decreased with higher bromide concentrations during conventional ozonation due to competitive ozone-consuming reactions, except for lamotrigine, due to the suspected reaction of HOBr/OBr- with the primary amine moieties. In addition to the abatement of micropollutants, the evolution of the two main transformation products (TPs) of hydrochlorothiazide (HCTZ) and tramadol (TRA), chlorothiazide (CTZ) and tramadol N-oxide (TRA-NOX), respectively, was assessed by chemical analysis and kinetic modeling. Both selected TPs were quickly formed initially to reach a maximum concentration followed by a decrease of their concentrations for longer contact times. For the studied conditions, the TP's concentrations at the outlet of the reactors ranged from 0 to 61% of the initial parent compound concentration, CTZ being a more persistent TP against further oxidation than TRA-NOX. Finally, it was demonstrated in both reactors that the formation of bromate (BrO3-), a potentially carcinogenic oxidation by-product, could be controlled by H2O2 addition with a general improvement on micropollutant abatement. Post-treatment by granular activated carbon (GAC) filtration enabled the reduction of micropollutants and TPs concentrations but no changes in bromate were observed. The combined algae assay showed that water quality was significantly improved after oxidation and GAC post-treatment, driven by the abatement of the spiked pesticides (diuron and atrazine).

Oxidation of cetirizine, fexofenadine and hydrochlorothiazide during ozonation: Kinetics and formation of transformation products.

The efficiency of wastewater ozonation for the abatement of three nitrogen-containing pharmaceuticals, two antihistamine drugs, cetirizine (CTR) and fexofenadine (FXF), and the diuretic drug, hydrochlorothiazide (HCTZ), was investigated. Species-specific second-order rate constants for the reactions of the molecular, protonated (CTR, FXF) or deprotonated (HCTZ) forms of these compounds with ozone were determined. All three compounds are very reactive with ozone (apparent second order rate constants at pH 7: kO3,pH7 = 1.7·10(5) M(-1)s(-1), 8.5·10(4) M(-1)s(-1) and 9.0·10(3) M(-1)s(-1) for CTR, HCTZ and FXF, respectively). Transformation product (TP) structures were elucidated using liquid chromatography coupled with high-resolution tandem mass spectrometry, including isotope-labeled standards. For cetirizine and hydrochlorothiazide 8 TPs each and for fexofenadine 7 TPs were identified. The main TPs of cetirizine and fexofenadine are their respective N-oxides, whereas chlorothiazide forms to almost 100% from hydrochlorothiazide. In the bacteria bioluminescence assay the toxicity was slightly increased only during the ozonation of cetirizine at very high cetirizine concentrations. The main TPs detected in bench-scale experiments were also detected in full-scale ozonation of a municipal wastewater, for >90% elimination of the parent compounds.

Exploring the Behaviour of Emerging Contaminants in the Water Cycle using the Capabilities of High Resolution Mass Spectrometry.

To characterize a broad range of organic contaminants and their transformation products (TPs) as well as their loads, input pathways and fate in the water cycle, the Department of Environmental Chemistry (Uchem) at Eawag applies and develops high-performance liquid chromatography (LC) methods combined with high-resolution tandem mass spectrometry (HRMS/MS). In this article, the background and state-of-the-art of LC-HRMS/MS for detection of i) known targets, ii) suspected compounds like TPs, and iii) unknown emerging compounds are introduced briefly. Examples for each approach are taken from recent research projects conducted within the department. These include the detection of trace organic contaminants and their TPs in wastewater, pesticides and their TPs in surface water, identification of new TPs in laboratory degradation studies and ozonation experiments and finally the screening for unknown compounds in the catchment of the river Rhine.

Chlorination of bisphenol A: non-targeted screening for the identification of transformation products and assessment of estrogenicity in generated water.

Besides the performance of water treatments on the removal of micropollutants, concern about the generation of potential biologically active transformation products has been growing. Thus, the detection and structural elucidation of micropollutants transformation products have turned out to be major issues to evaluate comprehensively the efficiency of the processes implemented for drinking water treatment. However, most of existing water treatment studies are carried out at the bench scale with high concentrations and simplified conditions and thus do not reflect realistic conditions. Conversely, this study describes a non-targeted profiling approach borrowed from metabolomic science, using liquid chromatography coupled to high-resolution mass spectrometry, in order to reveal potential chlorination products of bisphenol A (BPA) in real water samples spiked at 50µgL(-1). Targeted measurements first evidenced a fast removal of BPA (>99%) by chlorination with sodium hypochlorite (0.8mgL(-1)) within 10min. Then, the developed differential global profiling approach enabled to reveal 21 chlorination products of BPA. Among them, 17 were brominated compounds, described for the first time, demonstrating the potential interest of this innovative methodology applied to environmental sciences. In parallel to the significant removal of BPA, the estrogenic activity of water samples, evaluated by ER-CALUX assay, was found to significantly decrease after 10min of chlorination. These results confirm that chlorination is effective at removing BPA in drinking water and they may indicate that the generated compounds have significantly lower estrogenic activity.

Differential chemical profiling to identify ozonation by-products of estrone-sulfate and first characterization of estrogenicity in generated drinking water.

For a few years, the concern of water treatment companies is not only focused on the removal of target micropollutants but has been extended to the investigation of potential biologically active by-products generated during the treatment processes. Therefore, some methods dedicated to the detection and structural characterization of such by-products have emerged. However, most of these studies are usually carried out under simplified conditions (e.g. high concentration levels of micropollutants, drastic treatment conditions, use of deionized or ultrapure water) and somewhat unrealistic conditions compared to that implemented in water treatment plants. In the present study, a real field water sample was fortified at the part-per-billion level (50 µg L(-1)) with estrone-3-sulfate (E1-3S) before being ozonated (at 1 mg L(-1)) for 10 min. In a first step, targeted measurements evidenced a degradation of the parent compound (>80%) in 10 min. Secondly, a non-targeted chemical profiling approach derived from metabolomic profiling studies allowed to reveal 11 ozonation by-products, among which 4 were found predominant. The estrogenic activity of these water samples spiked with E1-3S before and after treatment was assessed by the ER-CALUX assay and was found to decrease significantly after 10 min of ozonation. Therefore, this innovative methodological strategy demonstrated its suitability and relevancy for revealing unknown compounds generated from water treatment, and permitted to generate new results regarding specifically the impact of ozonation on estrone-3-sulfate. These results confirm that ozonation is effective at removing E1-3S in drinking water and indicate that the by-products generated have significantly lower estrogenic activity.

Kinetic aspects and identification of by-products during the ozonation of bitertanol in agricultural wastewaters.

The degradation of bitertanol by ozone treatment is investigated. Solutions of bitertanol (8.4 µg mL(-1)) were prepared either by dissolution of the standard or by dilution of Gaucho Blé seed loading solution and then ozonated under different conditions. Evolution of the concentrations of bitertanol and its ozonation by-products in both solutions was monitored by HPLC-UV as a function of the treatment time for a concentration of 100 gm(-3) of ozone in the inlet gas. Bitertanol degradation was found to follow a pseudo-first order reaction in both cases. However, the rate of the reaction in diluted seed loading solution was much lower (0.19 vs. 0.27 min(-1) in standard solution) and was close to the reaction rate observed in the presence of a radical scavenger, tert-butanol (0.11 min(-1)). Thus, it may be suggested that additives present in the seed loading solution may play the role of radical scavengers. Study of ozone concentration in the inlet gas (from 25 to 100 gm(-3)) showed that ozone degradation is also a first-order reaction with respect to ozone. Four ozonation by-products were highlighted, collected and identified by HPLC coupled with an ion trap mass spectrometer using positive electrospray ionization mode. A degradation pathway of bitertanol was finally proposed.

Ozonation of imidacloprid in aqueous solutions: reaction monitoring and identification of degradation products.

This paper presents the degradation of imidacloprid by ozonation. Solutions of 39.0 µg/mL imidacloprid were prepared either by dissolution of standard or by dilution of Gaucho Blé(®) seed loading solution and then ozonated under different conditions. The concentration of imidacloprid and oxidation products in both solutions was monitored by HPLC-UV as a function of the treatment time for a concentration of 100g/m(3) of ozone in the inlet gas. No significant difference was observed: in both cases, imidacloprid degradation was a pseudo-first order reaction with similar reaction rates (0.129-0.147 min(-1)), degradation by-products with the same HPLC retention times were observed and their concentrations as a function of the treatment time followed a very similar trend. The study of ozone concentration in the inlet gas (from 25 to 100g/m(3)) showed that imidacloprid degradation is also a first-order reaction with respect to ozone. The ozonation by-products were then collected and identified by ESI(+)-MS. A degradation pathway of imidacloprid was finally proposed.

Development of a rapid determination of pesticides in coated seeds using a high-performance liquid chromatography-UV detection system.

For the determination of pesticides in coated seeds, this study compared two HPLC-UV methods, using a short column or a conventional column, as well as two extraction procedures, by ultrasonic extraction at room temperature or by pressurized liquid extraction (PLE). The comparison of selected column performances showed that the short column enabled the 3-fold reduction of analysis time (ca. 9 min vs 29 min) and eluent consumption (ca. 6.1 mL vs 20.8 mL) for the separation of five insecticides (bitertanol, fludioxonil, imidacloprid, metalaxyl-M and tefluthrin) and one bird repellent (anthraquinone) without altering peak resolutions. Recovery rates for pressurized liquid extraction at 120 degrees C were similar (between 84% and 102%) to those obtained by ultrasonication. Both methods were then applied for the extraction of loaded seeds. Rates for ultrasonic extraction at room temperature were lower (from 16% to 95%) than those observed for recovery tests, unlike PLE at 120 degrees C which showed good rates, ranging between 82% and 95%, for all the loaded pesticides.