Emerging Disinfection By-products Research Articles

Formation of the emerging disinfection byproducts halocyclopentadienes from phenolic compounds after chlorination

Halocyclopentadienes (HCPDs) are an emerging class of alicyclic disinfection by-products (DBPs) with high toxicity in disinfected drinking water. However, their potential precursors remain unclear, which hinders the understanding of their formation and further development of control strategies. In this study, two HCPDs, 1,2,3,4-tetrachloro-1,3-cyclopentadiene (TCC) and 1,2,3,4,5,5-hexachloro-1,3-cyclopentadiene (HCC), were identified in chlorinated lignin and tannic acid samples for the first time. The chlorination of four lignin-like and two tannic-like phenolic model compounds confirmed that guaiacol and digallic acid can produce HCPDs. According to their structures, ortho-substituents of phenolic compounds were speculated to be crucial for HCPDs formation. The simulated disinfection of catechol, 2-ethoxyphenol (2-EOP), 2-propoxyphenol (2-POP) and 3,4-dihydroxy-5-methoxybenzoic acid (DH-5-MBA) with different ortho-substituents demonstrated that three of these compounds can generate HCPDs, except catechol, which further indicates that ortho-substituents, such as the methoxy, ethoxy and propoxy groups, contribute to HCPDs generation. Guaiacol was the simplest compound for generating HCPDs, and possible formation pathways during chlorination were proposed. Seven hydroxy-chlorocyclopentadienes were tentatively identified and are likely important intermediates of HCPDs formation. Additionally, TCC and HCC were confirmed in tap water and chlorinated SRNOM samples with total concentrations up to 11.07 ng/L and 65.66 ng/L, respectively, further demonstrating the wide existence of HCPDs and their precursors. This study reports the clear precursors of HCPDs and provides a theoretical foundation for controlling HCPDs formation in disinfected drinking water.

Complexation with Metal Ions Affects Chlorination Reactivity of Dissolved Organic Matter: Structural Reactomics of Emerging Disinfection Byproducts.

Metal ions are liable to form metal-dissolved organic matter [dissolved organic matter (DOM)] complexes, changing the chemistry and chlorine reactivity of DOM. Herein, the impacts of iron and zinc ions (Fe3+ and Zn2+) on the formation of unknown chlorinated disinfection byproducts (Cl-DBPs) were investigated in a chlorination system. Fe3+ preferentially complexed with hydroxyl and carboxyl functional groups, while Zn2+ favored the amine functional groups in DOM. As a consequence, electron-rich reaction centers were created by the C-O-metal bonding bridge, which facilitated the electrophilic attack of α-C in metal-DOM complexes. Size-reactivity continuum networks were constructed in the chlorination system, revealing that highly aromatic small molecules were generated during the oxidation and decarbonization of metal-DOM complexes. Molecular transformation related to C-R (R represents complex sites) loss was promoted via metal complexation, including decarboxylation and deamination. Consequently, complexation with Fe3+ and Zn2+ promoted hydroxylation by the C-O-metal bonding bridge, thereby increasing the abundances of unknown polychlorinated Cl-DBPs by 9.6 and 14.2%, respectively. The study provides new insights into the regulation of DOM chemistry and chlorine reactivity by metal ions in chlorination systems, emphasizing that metals increase the potential health risks of drinking water and more scientific control standards for metals are needed.

Computational-aided analysis of the pathway and mechanism of dichloroacetonitrile formation from phenylalanine upon chloramination

Dichloroacetonitrile (DCAN) is an emerging disinfection by-product (DBP) that is widespread in drinking water. However, the pathway for DCAN formation from aromatic amino acids remains unclear, leading to a lack of an understanding of its explicit fate during chloramination. In this study, we investigated the specific formation mechanism of DCAN during the chloramination of phenylalanine based on reaction kinetics and chemical thermodynamics. The reason for differences between aldehyde and decarboxylation pathways was explained, and kinetic parameters of the pathways were obtained through quantum chemistry calculations. The results showed that the reaction rate constant of the rate-limiting step of the aldehyde pathway with 1.9 × 10−11 s−1 was significantly higher than that of decarboxylation (3.6 × 10−16 s−1 M−1), suggesting that the aldehyde pathway is the main reaction pathway for DCAN formation during the chloramination of phenylalanine to produce DCAN. Subsequently, theoretical calculations were performed to elucidate the effect of pH on the formation mechanism, which aligned well with the experimental results. Dehydrohalogenation was found to be the rate-limiting step under acidic conditions with reaction rate constants higher than those of the rate-limiting step (expulsion of amines) under neutral conditions, increasing the rate of DCAN formation. This study highlights the differences in DCAN formation between the decarboxylation and aldehyde pathways during the chloramination of precursors at both molecular and kinetic levels, contributing to a comprehensive understanding of the reaction mechanisms by which aromatic free amino acids generate DCAN.

Emerging disinfection byproducts 3-bromine carbazole induces cardiac developmental toxicity via aryl hydrocarbon receptor activation in zebrafish larvae

3-bromine carbazole (3-BCZ) represents a group of emerging aromatic disinfection byproducts (DBP) detected in drinking water; however, limited information is available regarding its potential cardiotoxicity. To assess its impacts, zebrafish embryos were exposed to 0, 0.06, 0.14, 0.29, 0.58, 1.44 or 2.88 mg/L of 3-BCZ for 120 h post fertilization (hpf). Our results revealed that ≥1.44 mg/L 3-BCZ exposure induced a higher incidence of heart malformation and an elevated pericardial area in zebrafish larvae; it also decreased the number of cardiac muscle cells and thins the walls of the ventricle and atrium while increasing cardiac output and impeding cardiac looping. Furthermore, 3-BCZ exposure also exhibited significant effects on the transcriptional levels of genes related to both cardiac development (nkx2.5, vmhc, gata4, tbx5, tbx2b, bmp4, bmp10, and bmp2b) and cardiac function (cacna1ab, cacna1da, atp2a1l, atp1b2b, atp1a3b, and tnnc1a). Notably, N-acetyl-L-cysteine, a reactive oxygen species scavenger, may alleviate the failure of cardiac looping induced by 3-BCZ but not the associated cardiac dysfunction or malformation; conversely, the aryl hydrocarbon receptor agonist CH131229 can completely eliminate the cardiotoxicity caused by 3-BCZ. This study provides new evidence for potential risks associated with ingesting 3-BCZ as well as revealing underlying mechanisms responsible for its cardiotoxic effects on zebrafish embryos.

Detection of emerging disinfection by-products by reversed-phase ion-pair chromatography electrospray-ionization mass spectrometry in drinking water

A sensitive and specific methodology that allows unequivocal identification of polydiallyldimethylammonium chloride (PDADMAC) disinfection by-product it monomer diallyldimethylammonium chloride (DADMAC) in drinking water samples based on reversed-phase ion-pair chromatography–electrospray ionization – mass spectrometry was optimized. Ion-pair reagent heptafluorobutyric (HFBA) was found to play a crucial role in the optimization of chromatographic parameters. The chromatographic retention time was optimized by varying concentrations of ion-pair reagent, flow rate, and column temperature to enable good chromatographic retention and sensitivity of disinfection by-product of interest DADMAC in positive electrospray ionization and using selected ion monitoring (SIM) mode. The method was validated and showed good linear regression of 0.99 and a detection limit of 0.008 mg L−1. The developed method was applied in a drinking water treatment plant operated by Mgeni Water in Durban, South Africa, to detect trace levels of the disinfection by-product of interest (DADMAC) at various points of the treatment process. The concentration of DADMAC in the water treatment plant ranged from not detected up to 0.65 mg L−1. In addition, this work through mass spectrometry provides the first evidence of ion-pairing (465 m/z peak) between DADMAC and the reagent, which is a confirmation of online derivatization. Also, the use of ion-pair prevented dimerization of DADMAC (286 m/z peak) inside the ionization chamber leading to high sensitivity.

Target analysis, occurrence and cytotoxicity of halogenated polyhydroxyphenols as emerging disinfection byproducts in drinking water

Halogenated aromatic disinfection byproducts (DBPs) in drinking water, such as halogenated phenols, have received widespread attention due to their high toxicity and ubiquitous occurrence in recent years. This study identified a group of emerging halogenated aromatic DBPs, known as halogenated polyhydroxyphenols (HPPs), and investigated their occurrence and cytotoxicity. We developed a highly sensitive solid-phase extraction ultra-performance liquid chromatography-tandem mass spectrometry (SPE-UPLC-MS/MS) method under multiple reaction monitoring (MRM) mode, with recoveries ranging from 86 to 115% and method detection limits (MDLs) ranging from 0.10 to 1.87 ng/L for the analysis of 15 HPPs. Eleven of these HPP DBPs were detected in collected drinking water samples using this method with detection frequencies ranging from 14 to 100% and a maximum concentration of 24 ng/L. The IC50 of the 15 HPPs in Chinese hamster ovary (CHO-K1) cells were ranged from 15.13 µM to 6.08×103 µM. The tested HPPs with -CHO substitution exhibited higher cytotoxicity compared to those with -COOH substitution. The TIC-Tox values of HPPs were calculated to be higher than those of HPs, indicating a potential necessity to pay attention to HPP DBPs. A quantitative structure−activity relationship (QSAR) model was developed for the cytotoxicity of HPPs, which was shown to be significantly associated with acid dissociation constant (pKa) and total valence connectivity (TVCon). To the best of our knowledge, this study reported the analysis, occurrence, and cytotoxicity of HPP DBPs in drinking water for the first time.

Oxidative stress as a key event in 2,6-dichloro-1,4-benzoquinone-induced neurodevelopmental toxicity

2,6-dichloro-1,4-benzoquinone (DCBQ) has been identified as an emerging disinfection byproducts (DBPs) in drinking water and has the potential to induce neurodevelopmental toxicity. However, there is rarely a comprehensive toxicological evaluation of the neurodevelopmental toxicity of DCBQ. Here, neural differentiating SH-SY5Y cells were used as an in vitro model. Our results have found that DCBQ has decreased cell viability and neural differentiation, generated higher level of reactive oxygen species (ROS), increased the percentage of apoptosis and lowered the level of mitochondrial membrane potential, suggesting the neurodevelopmental toxicity of DCBQ. In addition, antioxidant N-acetyl-L-cysteine (NAC) could significantly attenuate these DCBQ-induced neurotoxic effects, supporting our hypothesis that the neurodevelopmental toxicity may be related with oxidative stress induced by DCBQ. We further demonstrated that DCBQ-induced neurodevelopmental toxicity could promote the mitochondrial apoptosis pathway and inhibit the prosurvival PI3K/AKT/mTOR pathway through inducing ROS, which ultimately inhibited cell proliferation and induced apoptosis in neural differentiating SH-SY5Y cells. These findings have provided novel insights into the risk of neurodevelopmental toxic effects associated with DCBQ exposure, emphasizing the importance of assessing the potential neurodevelopmental toxicity of DBPs.

Emerging disinfection by-products formation of various molecular weight organic matter fractions in raw water contaminated with treated wastewater

Combining dissolved organic matter (DOM) in raw water (RW) with DOM in treated wastewater (TWW) can react with chlorine and pose emerging disinfection by-products (DBPs). This study evaluated DOM based on the molecular weight (MW) size fractionation, trihalomethane, iodinated-trihalomethane, haloacetonitrile, and trichloronitromethane formation potential (THMFP, I-THMFP, HANFP, and TCNMFP) of the RW from the U-Tapao Canal, Songkhla, Thailand and the RW mixed with TWW (RW + TWW) samples. The RW and RW + TWW were treated by coagulation with poly aluminum chloride. The DOM of RW and RW + TWW and their treated water was distributed most in the MW below 1 kDa. The MWs of 3–10 kDa and 1–3 kDa were the active DOM involved in the specific THMFP for the RW + TWW. The MW of < 1 kDa in the RW + TWW resulted in a slightly high specific I-THMFP and HANFP. The MW of 1 − 3 kDa in the coagulated samples had a high specific I-THMFP. The MW of > 10 kDa in the coagulated RW + TWW was a precursor for a particular HANFP. Monitoring systems for measuring the level of TWW mixed with RW and an effective process to enhance the efficiency of traditional water treatment must be set up to produce a consumer-safe water supply.

Determination of 13 halobenzoquinone disinfection by-products in drinking water using solid phase extraction-ultra performance liquid chromatography-triple quadrupole mass spectrometry

Disinfection of drinking water is critical to prevent waterborne diseases. An unexpected consequence of water disinfection is the formation of disinfection by-products by the interaction of disinfectants with organic matter (natural or anthropogenic) and halides, which present significant toxicological effects and carcinogenic risks. As an emerging disinfection by-product, halobenzoquinones (HBQs) have attracted increasing attention owing to their severe toxicity and high detection rates. The credible determination of HBQs is essential for further studies on their occurrence, toxicity, and control measures; however, HBQs are usually detected in drinking water at trace levels. Therefore, accurate and efficient analytical techniques are critical for HBQ determination and quantitation. In this study, a method based on solid phase extraction (SPE) combined with ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS) was developed to determine 13 HBQs, including six chlorobenzoquinones, six bromobenzoquinones, and one iodobenzoquinone, in drinking water. One-liter water samples were added with 2.5 mL of formic acid, and 500 mL of each sample was collected for further enrichment. Pretreatment optimization mainly focused on the SPE column, washing solvent, and nitrogen blowing temperature. After extraction using Plexa SPE columns (200 mg/6 mL), the samples were washed with ultrapure water containing 0.25% formic acid combined with 30% methanol aqueous solution containing 0.25% formic acid, eluted with 6 mL of methanol containing 0.25% formic acid, and then nitrogen blown at 30 ℃. The UPLC-MS/MS parameters were optimized by comparing the results of two reversed-phase columns (BEH C18 and HSS T3) and various concentrations of formic acid in the mobile phase, as well as by establishing the best instrumental conditions. The separation of 13 HBQs was performed using an HSS T3 column (100 mm×2.1 mm, 1.8 μm) via gradient elution with a mixture of 0.1% formic acid aqueous solution and methanol as the mobile phase for 16 min. The 13 HBQs were detected using a triple quadrupole mass spectrometer equipped with a negative electrospray ionization source (ESI-) in multiple reaction monitoring (MRM) mode. Matrix-matched calibration curves were used to quantify the HBQs owing to intense matrix inhibitory effects. The results reflected the good linear relationships of the 13 HBQs and yielded correlation coefficients (r) greater than 0.999. The method detection limits (MDLs, S/N=3) were 0.2-10.0 ng/L, while the method quantification limits (MQLs, S/N=10) were 0.6-33.0 ng/L. The recoveries of the 13 HBQs were 56%-88% at three spiked levels (10, 20, 50 ng/L), and the relative standard deviations (RSDs, n=6) were less than or equal to 9.2%. The optimization method was applied to analyze HBQs in five drinking water samples. Four HBQs, namely, 2,6-dichloro-1,4-benzoquinone (2,6-DCBQ), 2,5-dibromo-1,4-benzoquinone (2,5-DBBQ), 2,6-dibromo-1,4-benzoquinone (2,6-DBBQ), and 2,6-dibromo-3,5-dimethyl-1,4-benzoquinone (2,6-DBDMBQ), were detected in the samples with detection rates of 100%, 20%, 80%, and 20%, respectively. The most frequently detected HBQ, 2,6-DCBQ, also exhibited the highest content (15.0-56.2 ng/L). The method showed high sensitivity, stability, accuracy, and efficiency, rendering it suitable for the analysis of 13 HBQs in drinking water. Compared with previous methods that mainly focused on 2,6-DCBQ and 2,6-DBBQ, the developed method achieved higher throughput and enabled the simultaneous analysis of 13 HBQs. The method presented in this study provides an opportunity to explore different types and concentrations of HBQs in drinking water, offers a deeper understanding of the occurrence of HBQs, and facilitates further studies on the health risks and control measures of these compounds.

Uncovering an Emerging Group of Halogenated Nucleobase-Derived Disinfection Byproducts in Drinking Water: Prioritization of the Highly Cytotoxic 2-Chloroadenine.

Constant efforts have been devoted to exploring new disinfection byproducts in drinking water causally related to adverse health outcomes. In this study, five halogenated nucleobases were identified as emerging disinfection byproducts in drinking water, including 5-chlorouracil, 6-chlorouracil, 2-chloroadenine, 6-chloroguanine, and 5-bromouracil. We developed a solid phase extraction-ultraperformance liquid chromatography-tandem mass spectrometry method with the limits of detection (LOD) and recoveries ranging between 0.04-0.86 ng/L and 54-93%, respectively. The detection frequency of the five halogenated nucleobases ranged from 73 to 100% with a maximum concentration of up to 65.3 ng/L in the representative drinking water samples. The cytotoxicity of the five identified halogenated nucleobases in Chinese hamster ovary (CHO-K1) cells varied with great disparity, in which the cytotoxicity of 2-chloroadenine (IC50 = 9.4 μM) is appropriately three times higher than emerging DBP 2,6-dichloro-1,4-benzoquinone (IC50 = 42.4 μM), indicating the significant toxicological risk of halogenated nucleobase-DBPs. To the best of our knowledge, this study reports the analytical method, occurrence, and toxicity of halogenated nucleobase-DBPs for the first time. These findings will provide a theoretical basis for further research on probing the relationship between its mutagenicity and human health risk.

Degradation kinetics and formation of regulated and emerging disinfection by-products during chlorination of two expectorants ambroxol and bromhexine

Ambroxol hydrochloride (AMB) and bromhexine hydrochloride (BRO) are classic expectorants and bronchosecretolytic pharmaceuticals. In 2022, both AMB and BRO were recommended by medical emergency department of China to alleviate cough and expectoration for symptoms caused by COVID-19. The reaction characteristics and mechanism of AMB/BRO with chlorine disinfectant in the disinfection process were investigated in this study. The reaction of chlorine with AMB/BRO were well described by a second-order kinetics model, first-order in both AMB/BRO and chlorine. The second order rate reaction constant of AMB and BRO with chlorine at pH 7.0 were 1.15 × 102 M−1s−1 and 2.03 × 102 M−1s−1, respectively. During chlorination, a new class of aromatic nitrogenous disinfection by-products (DBPs) including 2-chloro-4, 6-dibromoaniline and 2, 4, 6-tribromoaniline were identified as the intermediate aromatic DBPs by gas chromatography-mass spectrometry. The effect of chlorine dosage, pH, and contact time on the formation of 2-chloro-4, 6-dibromoaniline and 2, 4, 6-tribromoaniline were evaluated. In addition, it was found that bromine in AMB/BRO were vital bromine source to greatly promote the formation of classic brominated DBPs, with the highest Br-THMs yields of 23.8% and 37.8%, respectively. This study inspired that bromine in brominated organic compounds may be an important bromine source of brominated DBPs.

Chlorinated nucleotides and analogs as potential disinfection byproducts in drinking water

Identification of emerging disinfection byproducts (DBPs) of health relevance is important to uncover the health risk of drinking water observed in epidemiology studies. In this study, mutagenic chlorinated nucleotides were proposed as potential DBPs in drinking water, and the formation and transformation pathways of these DBPs in chlorination of nucleotides were carefully investigated. A total of eleven chlorinated nucleotides and analogs were provisionally identified as potential DBPs, such as monochloro uridine/cytidine/adenosine acid and dichloro cytidine acid, and the formation mechanisms involved chlorination, decarbonization, hydrolysis, oxidation and decarboxylation. The active sites of nucleotides that reacted with chlorine were on the aromatic heterocyclic rings of nucleobases, and the carbon among the two nitrogen atoms in the nucleobases tended to be transformed into carboxyl group or be eliminated, further forming ring-opening or reorganization products. Approximately 0.2–4.0 % (mol/mol) of these chlorinated nucleotides and analogs finally decomposed to small-molecule aliphatic DBPs, primarily including haloacetic acids, trichloromethane, and trichloroacetaldehyde. Eight intermediates, particularly chlorinated imino-D-ribose and imino-D-ribose, were tentatively identified in chlorination of uridine. This study provides the first set of preliminary evidence for indicating the promising occurrence of chlorinated nucleotides and analogs as potential toxicological-relevant DBPs after disinfection of drinking water.

Hydroxylation of some emerging disinfection byproducts (DBPs) in water environment: Halogenation induced strong pH-dependency

In this work, the hydroxylation mechanisms and kinetics of some emerging disinfection byproducts (DBPs) have been systematically investigated through theoretical calculation methods. Five chlorophenols and eleven halogenated pyridinols were chosen as the model compounds to study their pH-dependent reaction laws in UV/H2O2 system. For the reactions of HO• with 37 different dissociation forms, radical adduct formation (RAF) was the main reaction pathway, and the reactivity decreased with the increase of halogenation degree. The kapp values (at 298 K) increased with the increase of pH from 0 to 10, and decreased with the increase of pH from 10 to 14. Compared with phenol, the larger the chlorination degree in chlorophenols was, the stronger the pH sensitivity of the kapp values; compared with chlorophenols, the pH sensitivity in halogenated pyridinols was further enhanced. As the pH increased from 2 to 10.5, the degradation efficiency increased at first and then decreased. With the increase of halogenation degree, the degradation efficiency range increased, the pH sensitivity increased, the optimal degradation efficiency slightly increased, and the optimal degradation pH value decreased. The ecotoxicity and bioaccumulation of most hydroxylated products were lower than their parental compounds. These findings provided meaningful insights into the strong pH-dependent hydroxylation of emerging DBPs on molecular level.

Methoxylated Modification of Glutathione-Mediated Metabolism of Halobenzoquinones In Vivo and In Vitro.

Xenobiotics were generally detoxified in organisms through interaction with endogenous molecules, which may also generate metabolites of increased toxicity. Halobenzoquinones (HBQs), a group of highly toxic emerging disinfection byproducts (DBPs), can be metabolized by reacting with glutathione (GSH) to form various glutathionylated conjugates (SG-HBQs). In this study, the cytotoxicity of HBQs in CHO-K1 cells showed a wavy curve as a function of increased GSH dosage, which was inconsistent with the commonly recognized progressive detoxification curve. We hypothesized that the formation and cytotoxicity of GSH-mediated HBQ metabolites contribute to the unusual wave-shaped cytotoxicity curve. Results showed that glutathionyl-methoxyl HBQs (SG-MeO-HBQs) were identified to be the primary metabolites significantly correlated with the unusual cytotoxicity variation of HBQs. The formation pathway was initiated by stepwise metabolism via hydroxylation and glutathionylation to produce detoxified hydroxyl HBQs (OH-HBQs) and SG-HBQs, followed by methylation to generate SG-MeO-HBQs of potentiated toxicity. To further verify the occurrence of the aforementioned metabolism in vivo, SG-HBQs and SG-MeO-HBQs were detected in the liver, kidney, spleen, testis, bladder, and feces of HBQ-exposed mice, with the highest concentration quantified in the liver. The present study supported that the co-occurrence of metabolism can be antagonistic, which enhanced our understanding of the toxicity and metabolic mechanism of HBQs.

Appraisal of machine learning techniques for predicting emerging disinfection byproducts in small water distribution networks

Monitoring emerging disinfection byproducts (DBPs) is challenging for many small water distribution networks (SWDNs), and machine learning-based predictive modeling could be an alternative solution. In this study, eleven machine learning techniques, including three multivariate linear regression-based, three regression tree-based, three neural networks-based, and two advanced non-parametric regression techniques, are used to develop models for predicting three emerging DBPs (dichloroacetonitrile, chloropicrin, and trichloropropanone) in SWDNs. Predictors of the models include commonly-measured water quality parameters and two conventional DBP groups. Sampling data of 141 cases were collected from eleven SWDNs in Canada, in which 70 % were randomly selected for model training and the rest were used for validation. The modeling process was reiterated 1000 times for each model. The results show that models developed using advanced regression techniques, including support vector regression and Gaussian process regression, exhibited the best prediction performance. Support vector regression models showed the highest prediction accuracy (R2 =0.94) and stability for predicting dichloroacetonitrile and trichloropropanone, and Gaussian process regression models are optimal for predicting chloropicrin (R2 =0.92). The difference is likely due to the much lower concentrations of chloropicrin than dichloroacetonitrile and trichloropropanone. Advanced non-parametric regression techniques, characterized by a probabilistic nature, were identified as most suitable for developing the predictive models, followed by neural network-based (e.g., generalized regression neural network), regression tree-based (e.g., random forest), and multivariate linear regression-based techniques. This study identifies promising machine learning techniques among many commonly-used alternatives for monitoring emerging DBPs in SWDNs under data constraints.

Formation of emerging disinfection byproducts from agricultural biomass-derived DOM: Overlooked health risk source

Carbon-derived dissolved organic matter (CDOM) are inevitably released to surface water during returning agricultural biomass carbon to farmland, which are potential precursors of disinfection byproducts (DBPs). In this study, CDOM was extracted from aerobic incineration (“OX”) and anoxic pyrolysis (“PY”) of three kinds of straw (wheat, corn, and rice), and the emerging DBPs from them were deciphered. The CDOM with molecular weight < 1 kDa in the OX and PY groups accounted for 53–87%, and it was higher in the PY group. A total 1343–2107 of CHO and 641–1761 of CHNO formulas were detected in the CDOM derived from the OX group, among which 74%-83% contained aromatic structures rich in oxygen containing groups. 1919–3289 of CHO and 785–1954 of CHNO formulas were observed in the PY group, and 77%-86% of them were lignins/CRAM-like compounds. Surprisingly, 765–2158 and 895–1648 of emerging DBPs were identified in the OX and PY groups, and the proportions of N-DBPs were 20.3–54.8% and 2.8–4.8%, respectively. Based on HOCl addition and Cl substitution mechanisms, the H/C ratios of the DBP precursors in the OX and PY groups were in the range of 0.2–1.5 and 0.6–2.0, respectively. The DBPs derived from the OX group exhibited higher cytotoxicity and genotoxicity due to the higher aromaticity and more N-DBPs. Thus, returning agricultural biomass carbon, particularly that produced by direct combustion, to farmland brought potential threat to drinking water safety.

Emerging Disinfection Byproduct 2,6-Dichlorobenzoquinone-Induced Cardiovascular Developmental Toxicity of Embryonic Zebrafish and Larvae: Imaging and Transcriptome Analysis.

Epidemiological studies have observed the potential association of water disinfection byproduct (DBP) exposure with cardiac defects. Aromatic DBPs represent a significant portion of total DBPs, but their effects on cardiovascular development are unclear. In this study, we examined the effects of an aromatic DBP, 2,6-dichlorobenzoquinone (DCBQ), on the cardiovascular development of zebrafish embryos. After exposure to 2, 4, and 8 μM DCBQ, morphological images of growing zebrafish embryos clearly showed cardiovascular malformation. Fluorescent images of transgenic zebrafish strains with fluorescently labeled heart and blood vessels show that DCBQ exposure resulted in deformed atrium-ventricle looping, degenerated abdomen and trunk vessels, pericardial edema, and decreased blood flow. Furthermore, the expression of the marker gene myl7 (essential for the differentiation and motility of cardiomyocytes) was inhibited in a dose-dependent manner by DCBQ exposure. Finally, transcriptome analysis found that in the 4 μM DCBQ exposure group, the numbers of differentially expressed genes (DEGs) were 113 (50 upregulated and 63 downregulated) at 24 hpf, 2123 (762 upregulated and 1361 downregulated) at 48 hpf, and 61 (11 upregulated and 50 downregulated) at 120 hpf; in the 8 μM DCBQ exposure group, the number of DEGs was 1407 (647 upregulated and 760 downregulated) at 120 hpf. The FoxO signaling pathway was significantly altered. The in vivo results demonstrate the effects of 2,6-DCBQ (0-8 μM) on cardiovascular development, contributing to the understanding of the developmental toxicity of aromatic DBP halobenzoquinones (HBQs).

Molecular insights of 2,6-dichlorobenzoquinone-induced cytotoxicity in zebrafish embryo: Activation of ROS-mediated cell cycle arrest and apoptosis.

2,6-dichloro-1,4-benzoquinone (2,6-DCBQ), as an emerging disinfection by-product, has been frequently detected in waters, posing potential health risk on public health. Although some studies have pointed out that 2,6-DCBQ exposure can induce cytotoxicity, limited information is available for underlying mechanism for 2,6-DCBQ-induced cytotoxicity. To explore this mechanism, we assessed the levels of reactive oxygen species (ROS), acridine orange (AO) staining, and the mRNA transcriptions of genes (Chk2, Cdk2, Ccna, Ccnb and Ccne) involved in cell-cycle and genes (p53, bax, bcl-2 and caspase 3) involved in apoptosis in zebrafish embryo, after exposed to different concentrations (10, 30, 60, 90 and 120 μg/L) of 2,6-DCBQ for 72 h. Our results indicated that 2,6-DCBQ exposure induced ROS generation and cell apoptosis, and disturbed the mRNA transcription of genes related to cell cycle and apoptosis in zebrafish embryo. Moreover, we also found that 30 ~ 60 μg/L 2,6-DCBQ is the important transition from cell-cycle arrest to cell apoptosis. These results provided novel insight into 2,6-DCBQ-induced cytotoxicity.

The convergence of 2,6-dichloro-1,4-benzoquinone in the whole process of lignin phenol precursor chlorination

The formation and decomposition of 2,6-dichloro-1,4-benzoquinone, an emerging disinfection byproduct (DBP), was studied in the chlorination of lignin phenol precursors. The results show that DCBQ and the related hydroxyl DCBQ (DCBQ-OH) acts as the intermediate products of the chlorination process of the three typical lignin phenol precursors (p-hydroxybenzoic acid, protocatechuic acid, and gallic acid). The contributions of lignin phenol precursors to the overall formation of the targeted DBPs were determined based on the observed abundances of individual lignin phenols and their DBP yields. DCBQ and DCBQ-OH were generated within 2–6 h, the relative abundance of the yields of mol carbon atoms in DCBQ corresponding to the mol carbon atoms in the three model precursors (DCBQ-C) was about 0.01%–14.37% under different pH conditions. With the chlorination reaction time increased (after two or four h), the concentrations of DCBQ and DCBQ-OH entirely decreased, and the decomposition of DCBQ do not follow a pseudo-first-order kinetics during chlorination. Conversely, the decomposition of DCBQ generated from p-hydroxybenzoic acid followed a pseudo-second-order kinetics. Moreover, the formation of trichloromethane (TCM), dichloroacetic acid (DCAA), and trichloroacetic acid (TCAA) was also detected during the chlorination. The contribution of the decomposed DCBQ was mainly to TCAA and the unknown DBPs within 2–12 h, and DCBQ decomposition pathway was affected by pH. Moreover, except for DCBQ/DCBQ-OH and TCM/HAAs, there were still 73.6%–92.41% unknown products (including non-halogenated aromatic DBPs and chlorine-substituted DBPs) needing to identify during the chlorination process for lignin phenols. Overall, revealing the formation and decomposition of DCBQ during the chlorination of lignin phenol precursors would contribute to the effective development of drinking water treatment processes for the removal of highly toxic intermediates generated during disinfection.

Untargeted metabolomics of human keratinocytes reveals the impact of exposure to 2,6-dichloro-1,4-benzoquinone and 2,6-dichloro-3-hydroxy-1,4-benzoquinone as emerging disinfection by-products

IntroductionThe 2,6-dichloro-1,4-benzoquinone (DCBQ) and its derivative 2,6-dichloro-3-hydroxy-1,4-benzoquinone (DCBQ-OH) are disinfection by-products (DBPs) and emerging pollutants in the environment. They are considered to be of particular importance as they have a high potential of toxicity and they are likely to be carcinogenic.ObjectivesIn this study, human epidermal keratinocyte cells (HaCaT) were exposed to the DCBQ and its derivative DCBQ-OH, at concentrations equivalent to their IC20 and IC50, and a study of the metabolic phenotype of cells was performed.MethodsThe perturbations induced in cellular metabolites and their relative content were screened and evaluated through a metabolomic study, using 1H-NMR and MS spectroscopy.ResultsChanges in the metabolic pathways of HaCaT at concentrations corresponding to IC20 and IC50 of DCBQ-OH involved the activation of cell membrane α-linolenic acid, biotin, and glutathione and deactivation of glycolysis/gluconeogenesis at IC50. The changes in metabolic pathways at IC20 and IC50 of DCBQ were associated with the activation of inositol phosphate, pertaining to the transfer of messages from the receptors of the membrane to the interior as well as with riboflavin. Deactivation of biotin metabolism was recorded, among others. The cells exposed to DCBQ exhibited a concentration-dependent decrease in saccharide concentrations. The concentration of steroids increased when cells were exposed to IC20 and decreased at IC50. Although both chemical factors stressed the cells, DCBQ led to the activation of transporting messages through phosphorylated derivatives of inositol.ConclusionOur findings provided insights into the impact of the two DBPs on human keratinocytes. Both chemical factors induced energy production perturbations, oxidative stress, and membrane damage.