Effects of exposure to 3,6-DBCZ on neurotoxicity and AhR pathway during early life stages of zebrafish (Danio rerio).

Polyhalogenated carbazoles (PHCZs) are emerging environmental pollutants, yet limited information is available on their embryotoxicity and neurotoxicity. Therefore, the current work was performed to investigate the adverse effects of 3,6-dibromocarbazole (3,6-DBCZ), a typical PHCZs homolog, on the early life stages of zebrafish larvae. It revealed that the 96-hour post-fertilization (hpf) median lethal concentration (LC50) value of 3,6-DBCZ in zebrafish larvae was determined to be 0.7988 mg/L. Besides, 3,6-DBCZ reduced survival rates at concentrations ≥ 1 mg/L and decreased hatching rates at ≥ 0.25 mg/L at 48 hpf. In behavior tests, it inhibited locomotor activities and reduced the frequency of recorded acceleration states in response to optesthesia (a sudden bright light stimulus) at concentrations ≥ 160 µg/L. Meanwhile, 3,6-DBCZ exposure decreased the frequency of recorded acceleration states in the startle response (tapping mode) at concentrations ≥ 6.4 µg/L. Pathologically, with the transgenic zebrafish model (hb9-eGFP), we observed a strikingly decreased axon length and number in motor neurons after 3,6-DBCZ treatment, which may be ascribed to the activation of the AhR signaling pathway, as evidenced by the molecular docking analysis and Microscale thermophoresis (MST) assay suggested that 3,6-DBCZ binding to AhR-ARNT2 compound proteins. Through interaction with AhR-ARNT, a striking reduction of the anti-oxidative stress (sod1/2, nqo1, nrf2) and neurodevelopment-related genes (elavl3, gfap, mbp, syn2a) were observed after 3,6-DBCZ challenge, accompanied by a marked increased inflammatory genes (TNFß, IL1ß, IL6). Collectively, our findings reveal a previously unrecognized adverse effect of 3,6-DBCZ on zebrafish neurodevelopment and locomotor behaviors, potentially mediated through the activation of the AhR pathway. Furthermore, it provides direct evidence for the toxic concentrations of 3,6-DBCZ and the potential target signaling in zebrafish larvae, which may be beneficial for the risk assessment of the aquatic ecosystems.

Lycopene abolishes typical polyhalogenated carbazoles (PHCZs)-induced hepatic injury in yellow catfish (Pelteobagrus fulvidraco): Involvement of ROS/PI3K-AKT/NF-κB signaling.

Aquatic ecosystems are being more contaminated with polyhalogenated carbazoles (PHCZs), which raising concerns about their impact on aquatic organisms. Lycopene (LYC) exhibits several beneficial properties for fish via enhance antioxidant defenses and improve immunity. In this study, we attempted to investigate the hepatotoxic effects of typical PHCZs 3, 6-dichlorocarbazole (3,6-DCCZ) and the protective mechanisms of LYC. In this study, we found that yellow catfish (Pelteobagrus fulvidraco) exposure to 3,6-DCCZ (1.2 mg/L) resulted in hepatic inflammatory infiltration and disordered hepatocyte arrangement. Besides, we observed that 3,6-DCCZ exposure resulted in hepatic reactive oxygen species (ROS) overproduction and excessive autophagosome accumulation, accompanied with inhibition of phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) pathway. Subsequently, we confirmed that 3,6-DCCZ exposure triggered hepatic uncontrolled inflammatory response via activation of nuclear factor-κB (NF-κB) pathway, along with decreased plasma complement C3 (C3) and complement C4 (C4) levels. Meanwhile, yellow catfish exposed to 3,6-DCCZ exhibit an increased hepatic apoptosis phenomenon, as evidenced by the elevated number of positive TUNEL cells and upregulated expression of caspase3 and cytochrome C (CytC). In contrast, LYC treatment could alleviate the 3,6-DCCZ-induced pathological changes, hepatic ROS accumulation, autophagy, inflammatory response and apoptosis. To sum up, this study provided the demonstration that LYC exerts hepatoprotective effects to alleviate 3,6-DCCZ-induced liver damage by inihibiting ROS/PI3K-AKT/NF-κB signaling in yellow catfish.

Production of Polyhalogenated Carbazoles in Marine Red Alga Corallina officinalis: A Possible Natural Source.

Polyhalogenated carbazoles (PHCZs) have been increasingly detected in the environment as a result of anthropogenic and natural origin. However, it is unclear how PHCZs are naturally produced. In this study, the formation of PHCZs from bromoperoxidase (BPO)-mediated halogenation of carbazole was investigated. A total of six PHCZs were identified in reactions under different incubation conditions. The presence of Br- significantly influenced the formation of PHCZs. The products were first dominated by 3-bromocarbazole and then 3,6-dibromocarbazole as the reactions proceeded. Both bromo- and chlorocarbazoles were identified in the incubations with trace Br-, suggesting the co-occurrence of BPO-catalyzed bromination and chlorination. However, BPO-catalyzed chlorination of carbazole was much weaker than that of bromination. The formation of PHCZs could be attributable to the halogenation of carbazole by reactive halogen species generated from BPO-catalyzed oxidation of Br- and Cl- by H2O2. The halogenation was found to follow a successive substitution order of C-3, C-6, and C-1 on the carbazole ring, forming 3-, 3,6-, and 1,3,6-isomers. Similar to the incubation experiments, six PHCZs were for the first time detected in red algal samples collected from the South China Sea, China, suggesting the biogenesis of PHCZs in marine red algae. Given the widespread distribution of red algae in the marine environment, BPO-catalyzed halogenation of carbazole may be a natural origin for PHCZs.

Polyhalogenated carbazoles induce hepatic metabolic disorders in mice via alteration in gut microbiota.

Polyhalogenated carbazoles (PHCZs) have been widely accepted as emerging pollutants, whereas their ecological and health risks remain uncertain. Herein, female and male Sprague-Dawley (SD) mice were treated with four typical PHCZs to investigate their negative consequences, along with alternations in gut microbiota to indicate underlying mechanisms. In female mice, the relative liver weight ratio increased after four PHCZs exposure; 2-bromocarbazole (2-BCZ) increased urine glucose level; 3-bromocarbazole (3-BCZ) decreased the glucose and total cholesterol levels; 3,6-dichlorocarbazole (3,6-DCCZ) decreased glucose level. The only disturbed biochemical index in male mice was the promoted alkaline phosphatase (ALP) level by 3,6-DCCZ. We also found that the differential blood biochemical indices were correlated with gut microbiota. 3-BCZ and 3,6-DCCZ altered Bacteroidetes and Proteobacteria phyla in female and male mice, which were correlated with metabolic disorders. Our findings demonstrated the correlation between PHCZs induced potential hepatotoxicity and metabolic disorders may be due to their dioxin-like potentials and endocrine disrupting activities, and the gender differences might result from their estrogenic activities. Overall, data presented here can help to evaluate the ecological and health risks of PHCZs and reveal the underlying mechanisms.

PM2.5-bound polyhalogenated carbazoles (PHCZs) in urban Beijing, China: Occurrence and the source implication.

Polyhalogenated carbazoles (PHCZs) are recently raising much attention due to their toxicity and ubiquitous environmental distribution. However, little knowledge is known about their ambient occurrences and the potential source. In this study, we developed an analytical method based on GC-MS/MS to simultaneously determine 11 PHCZs in PM2.5 from urban Beijing, China. The optimized method provided low method limit of quantifications (MLOQs, 1.45-7.39 fg/m3) and satisfied recoveries (73.4%-109.5%). This method was applied to analyze the PHCZs in the outdoor PM2.5 (n = 46) and fly ash (n = 6) collected from 3 kinds of surrounding incinerator plants (steel plant, medical waste incinerator and domestic waste incinerator). The levels of ∑11PHCZs in PM2.5 ranged from 0.117 to 5.54 pg/m3 (median 1.18 pg/m3). 3-chloro-9H-carbazole (3-CCZ), 3-bromo-9H-carbazole (3-BCZ), and 3,6-dichloro-9H-carbazole (36-CCZ) were the dominant compounds, accounting for 93%. 3-CCZ and 3-BCZ were significantly higher in winter due to the high PM2.5 concentration, while 36-CCZ was higher in spring, which may be related to the resuspending of surface soil. Furthermore, the levels of ∑11PHCZs in fly ash ranged from 338 to 6101 pg/g. 3-CCZ, 3-BCZ and 36-CCZ accounted for 86.0%. The congener profiles of PHCZs between fly ash and PM2.5 were highly similar, indicating that combustion process could be an important source of ambient PHCZs. To the best of our knowledge, this is the first research providing the occurrences of PHCZs in outdoor PM2.5.

Study of the scavenger and vector roles of microplastics for polyhalocarbazoles under simulated gastric fluid conditions.

Microplastics entering the digestive system of living organisms can serve as a carrier of hydrophobic organic pollutants (HOPs), increasing their exposure levels and the health risks they pose to both humans and animals. The desorption kinetics of six polyhalocarbazoles (PHCZs) from 5 mm and 0.15 mm polypropylene (PP) and polyvinyl chloride (PVC) microplastic particles were assessed using a combined microplastics and food system, representing the gastric system of vertebrates and invertebrates. Results showed that the chemical transfer of PHCZs is biphasic and reversible, with rapid exchange occurring within 2-48 h, followed by a period of slow transfer, which continues for weeks to months. The desorption capacity of PHCZs loaded on 0.15 mm microplastic particles was greater than that of 5 mm particles. The bioavailability percentage of PHCZ congeners for PP (24.2%-65.3%) and PVC (43.5%-57.2%) in the vertebrate fluid system were all lower than those in the invertebrate system (34.2%-70.7% for PP and 56.3%-72.7% for PVC, respectively). These findings indicate that physiological conditions, such as polarity, ingestion fluid, and microplastic affect the desorption of PHCZs from microplastics. In addition, desorption from PP was inhibited by the presence of foodstuff loaded with PHCZs due to competition, while desorption from PVC was not significantly affected by the presence of PHCZs contaminant food. Microplastics could provide a cleaning function in gastric fluid systems containing contaminated foodstuff, especially PP, which was capable of competitive adsorption of PHCZs from food. Few investigations have focused on the adverse effects of microplastic ingestion on human health, particularly in their role as vectors for HOPs, compared to other routes of exposure and transport. Therefore, these findings provide valuable insight into the health risks associated with dietary intake of microplastics and HOPs.

Isolation and identification of 1,3,6,8-tetrabromocarbazole - degrading bacteria.

Halogenated carbazoles are a new class of persistent organic pollutants with dioxin-like toxicity, and this study focused on the microbial degradation of 1,3,6,8-tetrabromocarbazole. In this study, a novel 1,3,6,8-tetrabromocarbazole (1,3,6,8-TBCZ) degrading strain TB-1 was isolated from contaminated soil and identified as Achromobacter sp. based on its 16S rRNA gene sequence analysis, morphological, physiological, and biochemical characteristics. The soil sample was collected from a pharmaceutical factory in Suzhou, China. The strain was able to effectively degrade 1 mg L-1 1,3,6,8-TBCZ in 7 d at pH 7.0 and 30 °C with 80% degradation rate. During the process, the intermediate metabolites were identified as Tribromocarbazole, dibromocarbazole and bromocarbazole via gas chromatography mass spectrometry (GC-MS). The results indicated that strain TB-1 may contribute to the bioremediation of polyhalogenated carbazoles (PHCs) in contaminated environment.

Determination of polyhalogenated carbazoles in waters at low nanogram-per-liter concentrations with solid-phase disk extraction.

Polyhalogenated carbazoles, a class of emerging contaminants with persistence and dioxin-like toxicity, have received increasing attention in recent years. In this study, a simple, rapid, sensitive, and high throughput method based on solid-phase disk extraction and gas chromatography-mass spectrometry was described for the determination of polyhalogenated carbazoles in low nanogram-per-liter range in water samples. The proposed solid-phase disk extraction method was initially optimized, and the optimum experimental conditions found were 1 L water sample (pH 6-9) extracted and enriched by Empore 3-stn octadecyl disk at flow rate of 5 to 50 mL/min and eluted by 5 mL of acetone and 3 × 10 mL methylene dichloride. The linearity of the method ranged from 0.2 to 50 ng/L for carbazole and 11 polyhalogenated carbazoles, with correlation coefficients ranging from 0.9951 to 0.9996. The limits of detection were in the low nanogram per liter level, ranging from 0.018 to 0.12 ng/L. Finally, the optimized method was applied for determining trace levels of carbazole and 11 polyhalogenated carbazoles in tap water and seawater samples with good recovery of 86.6-112.8%. Carbazole and 3-7 polyhalogenated carbazoles were detected, and 3,6-dichlorocarbazole was the predominant congener both in tap water and seawater.

Analysis of polyhalogenated carbazoles in sediment using liquid chromatography-tandem mass spectrometry.

The purpose of this study was to develop a novel and sensitive method for the analysis of carbazole and polyhalogenated carbazoles (PHCs) in sediment using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Briefly, 5.0 g of freeze-dried sediment samples were extracted with dichloromethane using pressurized liquid extraction (PLE). The extract was purified with Florisil solid phase extraction cartridge, filtered through 0.22 µm polytetrafluoroethylene filter using a glass syringe, followed by LC-MS/MS analysis. Besides parameters for LC-MS/MS analysis, sample preparation procedures (including solvents for PLE, sorbents for cleanup, and filters for sample filtration) were optimized. The limits of detection and limits of quantification of target compounds were in the ranges of 3.0 × 10-3 to 0.22 ng g-1 dry weight (d.w.) and 1.0 × 10-2 to 0.75 ng g-1 d.w., respectively. The recoveries of target compounds in the spiked sediments at 2.0 ng g-1 d.w. and 10 ng g-1 d.w. were 64.8-91.8% and 70.9-124.7%, respectively, with relative standard deviations being less than 13.2%. Except that 36-BCZ had positive matrix effects of 63.3%, the sediment matrices generally displayed low or medium negative matrix effects on the other target compounds during LC-MS/MS analysis. The developed method was applied in the analysis of carbazoles and PHCs in sediment samples from Jiulong River, Fujian, China and all the target compounds were detected in the samples.

Polyhalogenated Carbazoles in Surface Sediment from Sanmen Bay, East China Sea: Spatial Distribution and Congener Profile.

Polyhalogenated carbazoles (PHCZs) have recently emerged as a group of halogenated pollutants with broad occurrences and bioaccumulation potential in aquatic systems. However, investigations on their occurrences in coastal waters remain very limited. In the present study we investigated PHCZs in surface sediment collected from 29 sites in Sanmen Bay, East China Sea. The results demonstrated a universal presence of PHCZs in sediment, with concentrations of ∑PHCZs (including all congeners) ranging from 7.7 to 17.5 ng/g dry weight (median: 11.3 ng/g dw). The PHCZ congener composition profile revealed a dominance of 3,6-dichlorocarbazole (36-CCZ) with comparable concentration with that of carbazole. Given that PHCZs are widely distributed in Sanmen Bay sediment and their concentrations rivaled other well-known persistent organic pollutants in the same area, this group of halogenated pollutants merits additional investigations of their potential risks to the studied aquatic system, as well as other important watersheds.

Combined toxic effects of polystyrene microplastics and 3,6-dibromocarbazole on zebrafish (Danio rerio) embryos.

Microplastics (MPs) and polyhalogenated carbazoles (PHCZs) are widely detected in the aquatic environment, and their ecological risks have become a research focus. Although there is an extensive co-distribution of MPs and PHCZs, their combined toxicity to aquatic organisms is still unclear. This study investigated the toxic effects of polystyrene microplastics (PS-MPs) and 3,6-dibromocarbazole (3,6-DBCZ) on zebrafish embryos by individual/combined exposure. This study showed that individual or combined exposure of PS-MPs (10 mg/L) and 3,6-DBCZ (0.5 mg/L) could significantly increase the rate of zebrafish embryo deformity, whereas no significant effect was observed on mortality and hatching rate. Furthermore, exposure to 3,6-DBCZ or PS-MPs increased reactive oxygen species (ROS) levels in zebrafish embryos, and the resulting oxidative stress induced apoptosis. Comparably, the levels of oxidative stress and apoptosis in zebrafish embryos were significantly reduced with the combined exposure of 3,6-DBCZ and PS-MPs. These observations suggest that the combined exposure of 3,6-DBCZ and PS-MPs has an antagonistic effect on oxidative stress and apoptosis. Fluorescence PS-MPs tracing and 3,6-DBCZ enrichment analysis showed that, with the protection of chorion, the entry of PS-MPs (5 and 50 µm) into the embryonic stage (55 hpf) of zebrafish was prevented. Moreover, after exposure for 96-144 hpf, PS-MPs served as a carrier to promote the 3,6-DBCZ accumulation and its dioxin-like toxicity in zebrafish larvae through ingestion. Compared with 5-µm PS-MPs, 50-µm PS-MPs promoted higher accumulation and dioxin-like toxicity of 3,6-DBCZ in zebrafish larvae. These findings provide that MPs can be used as an important carrier of PHCZs, influencing their toxicity and bioaccumulation in the organisms.

Environmental behavior and risk of the emerging organic contaminants halogenated carbazoles in chemical industrial park clusters.

Polyhalogenated carbazoles (PHCZs) are emerging organic contaminants and have attracted extensive concern because of their widespread occurrence and dioxin-like toxicity. However, the distribution characteristics, environmental behavior, and fate of PHCZs are still poorly understood. In this study, 74 composite environmental samples from 21 Chinese cities were collected around industrial parks in the Yangtze River Delta. The PHCZ concentration ranges in sediment and soil samples were 12.7-5.21 × 103 and 34.6-1.81 × 103 ng/g, respectively, which is equivalent to or higher than those of well-known persistent organic pollutants in the similar areas. The dominant congeners of PHCZs in sediment and soil were 3-chlorocarbazole and 3,6-dichlorocarbazole. Industrial emissions, especially from printing and dyeing textiles, were the main contributors to the high PHCZ environmental concentrations. Potential toxic effects of the PHCZs were evaluated using the toxic equivalent (TEQ) method. The TEQs of PHCZs in sediment and soil were up to 550 and 554 pg TEQ/g dry weight, respectively. The estimated TEQ value of sediment and soil exceeded the corresponding safety guideline, which indicated that PHCZs in the Yangtze River Delta posed high health risks. This study provides an important theoretical basis for controlling and reducing the ecological risks of PHCZs in the chemical industry. At the same time, it also provides reference for the priority control and revision of discharge standards for PHCZs in sewage treatment plants in future.

Tissue-specific accumulation, depuration and histopathological effects of 3,6-dichlorocarbazole and 2,7-dibromocarbazole in adult zebrafish (Danio rerio).

Although polyhalogenated carbazoles have been detected with increasing frequency in aquatic ecosystems, their bioaccumulation in fish and corresponding pathological effects related to bioaccumulation are still unclear. Here, we investigated the tissue-specific accumulation, depuration, and histopathological effects of two typical PHCZs, 3,6-dichlorocarbazole (36-CCZ) and 2,7-dibromocarbazole (27-BCZ), in adult zebrafish at three levels (0, 0.15 µg/L (5 × environmentally relevant level), and 50 µg/L (1/10 LC50). The lowest concentrations of 36-CCZ (1.2 µg/g ww) and 27-BCZ (1.4 µg/g ww) were observed in muscle, and the greatest concentrations of 36-CCZ (3.6 µg/g ww) and 27-BCZ (4 µg/g ww) were detected in intestine among the tested tissues. BCFww of 36-CCZ and 27-BCZ in zebrafish ranged from 172.9 (muscle) to 606.6 (intestine) and 285.2 (muscle) to 987.5 (intestine), respectively, indicating that both 36-CCZ and 27-BCZ have high potential of bioaccumulation in aquatic system. The 0.15 µg/L level of 36-CCZ or 27-BCZ caused lipid accumulation in liver, while 50 µg/L of 36-CCZ or 27-BCZ induced liver lesions such as fibrous septa, cytolysis, and nuclear dissolution. Brain damage such as multinucleated cells and nuclear solidification were only observed at 50 µg/L of 27-BCZ. This study provided valuable information in assessing the health and ecological risks of 36-CCZ and 27-BCZ.

Occurrence of polyhalogenated carbazoles and the combined effects with heavy metals on variation in bacterial communities in estuarine sediments.

Carbazole (CZ) and eight polyhalogenated carbazoles (PHCZs) were quantified by GC-MS in sediments of 12 estuaries, the interface linking large industrial and living areas to the Bohai Sea, China. These pollutants, heavy metals, and environmental factors caused integrated exposure to sediment bacteria. Four PHCZ congeners were detectable, with ΣPHCZs ranging from 0.56 to 15.94 ng/g dw. The dominant congeners were 3,6-dichlorocarbazole (36-CCZ) and 3-chlorocarbazole (3-CCZ), with a mean contribution of 72.6 % and 20.2 %. Significant positive correlations were found between 36-CCZ and both total organic carbon and heavy metals. Redundancy analysis of microbial variation implicated no impacts from PHCZs. Correlation analysis demonstrated an increase in abundance of Rhodocyclaceae but a decrease in Bacteroides-acidifaciens-JCM-10556 with presence of PHCZs, suggesting that these bacteria can be used as potential contamination indicators. The combined exposure of heavy metals, nutrients, and PHCZs may also increase toxicity and biological availability, adversely affecting the ecosystem and human health.

Spatial distribution characteristics of carbazole and polyhalogenated carbazoles in water column and sediments in the open Western Pacific Ocean.

Polyhalogenated carbazoles (PHCZs), an emerging persistent halogenated organic pollutant, have been detected in the environment. However, our understanding of PHCZs in the ocean remains limited. In this study, 47 seawater samples (covering 50 - 4000 m) and sediment samples (49 surface and 3 cores) were collected to investigate the occurrence and spatial distribution patterns of carbazole and its halogenated derivants (CZDs) in the Western Pacific Ocean. In seawater, the detection frequencies of CZ (97.87%) and 3-CCZ (57.45%) were relatively high. In addition, the average concentration of ΣPHCZs in the upper water (< 150 m, 0.23 ± 0.21 ng/L) was significantly lower than that in the deep ocean (1000 - 4000 m, 0.65 ± 0.56 ng/L, P < 0.05), which may indicate the vertical transport of PHCZs in the marine environment. The concentration of ΣCZDs in surface sediment ranges from 0.46 to 6.48 ng/g (mean 1.54 ng/g), among which CZ and 36-CCZ were the predominant components. Results from sediment cores demonstrate a noteworthy negative correlation between the concentration of CZDs and depth, indicating the ongoing natural degradation process occurring in sediment cores over a long period. This study offers distinctive insights into the occurrence, composition, and vertical features of CZDs in oceanic environments.

Unintended polyhalogenated carbazole production during advanced oxidation of coking wastewater.

Polyhalogenated carbazoles (PHCZs) are emerging as dioxin-like global pollutants, yet their environmental origins are not fully understood. This study investigates the application of the Fenton process in coking wastewater treatment, focusing on its dual role in carbazole removal and unintended PHCZ formation. The common halide ions (Cl- and Br-) in coking wastewater, especially Br- ions, exerted a notable impact on carbazole removal. Particularly, the influence of Br- ions was more significant, not only enhancing carbazole removal but also shaping the congener composition of PHCZ formation. Elevated halide ion concentrations were associated with the heightened formation of higher halogenated carbazoles. The Fenton reagent dosage ratio was identified as a crucial factor affecting the congener composition of PHCZs and their toxic equivalency value. The coexisting organic substance (i.e., phenol) in coking wastewater was observed to inhibit PHCZ formation, likely through competitive reactions with carbazole. Intriguingly, ammonium (NH4+) facilitated the generation of higher and mixed halogenated carbazoles, possibly due to the generation of nitrogen-containing brominating agents with stronger bromination capacity. This study underscores the importance of a comprehensive assessment, considering both substrate removal and potential byproduct formation, when employing the Fenton process for saline wastewater treatment.

Chronic exposure to 3,6-dichlorocarbazole exacerbates non-alcoholic fatty liver disease in zebrafish by disrupting lipid metabolism and inducing special lipid biomarker accumulation.

Most previous studies have focused primarily on the adverse effects of environmental chemicals on organisms of good healthy. Although global prevalence of non-alcoholic fatty liver disease (NAFLD) has reached approximately 25%, the impact of environmentally persistent organic chemicals on organisms with NAFLD is substantially unknown. Polyhalogenated carbazoles (PHCZs) as emerging contaminants have been frequently detected in the environment and organisms. In this study, we investigated the impact of the most frequently detected PHCZs, 3,6-dichlorocarbazole (36-CCZ), on zebrafish with high-fat diet (HFD)-induced NAFLD. After 4 weeks exposure to environmentally relevant concentrations of 36-CCZ (0.16-0.45 µg/L), the accumulation of lipid in zebrafish liver dramatically increased, and the transcription of genes involved in lipid synthesis, transport and oxidation was significantly upregulated, demonstrating that 36-CCZ had exacerbated the NAFLD in zebrafish. Lipidomic analysis indicated that 36-CCZ had significantly affected liver lipid metabolic pathways, mainly including glycerolipids and glycerophospholipids. Additionally, fifteen lipids were identified as potential lipid biomarkers for 36-CCZ exacerbation of NAFLD, including diacylglycerols (DGs), triglycerides (TGs), phosphatidylcholines (PCs), phosphatidylethanolamines (PEs), phosphatidic acid (PA), and phosphatidylinositol (PI). These findings demonstrate that long-term exposure to 36-CCZ can promote the progression of NAFLD, which will contribute to raising awareness of the health risks of PHCZs.

Development of a three-dimensional porous ionic liquid-chitosan-graphene oxide aerogel for efficient extraction and detection of polyhalogenated carbazoles in sediment samples.

The three-dimensional porous ionic liquid-chitosan-graphene oxide aerogel (IL-CS-GOA) monolithic adsorbent with a through-hole structure was prepared using natural chitosan (CS) as the skeletal framework, graphene oxide (GO) as the support to provide mechanical strength, and ionic liquid (IL) as the porogen and modifier. The resulting IL-CS-GOA demonstrated a fluffy and porous structure with various pore sizes and excellent regeneration capability (over six cycles). Its specific surface area exceeded that of CS-GOA and IL-GOA by more than 7 times, enhancing its polyhalogenated carbazoles (PHCZs) adsorption capacity. Within 5 min, IL-CS-GOA (1.0 mg) exhibited adsorption amounts of 539 ng mg-1 for 3-bromocarbazole (3-BCZ), 716 ng mg-1 for 2,7-dibromocarbazole (2,7-BCZ), and 798 ng mg-1 for 1,3,6,8-tetrabromocarbazole (1,3,6,8-BCZ), showcasing its rapid mass transfer and high adsorption capabilities. IL-CS-GOA was utilized as the adsorbent for glass dropper extraction (GDE) in conjunction with gas chromatography-mass spectrometry (GC-MS/MS), to develop a highly efficient and accurate method for determining PHCZs in sediments. Under optimal conditions, the established method exhibited a wide linear range (0.4-250 ng g-1, r ≥ 0.9990), low detection limits (0.04-0.24 ng g-1), and satisfactory recoveries (80.5 %-93.8 %), enabling the accurate and rapid detection of PHCZs in sediment samples. This study presents a novel approach for creating three-dimensional porous aerogels, introduces a new form of sample pretreatment using GDE with a monolithic adsorbent, and offers a new method for the determination of PHCZs in environmental matrices.

Analysis of polyhalogenated carbazoles and two related compounds in earthworms using a modified QuEChERS method with GC/MS and GC/MS/MS.

A precise analytical method based on QuEChERS has been proposed for the concurrent determination of 11 polyhalogenated carbazoles (PHCZs), benzocarbazole (BZCZ), and 9H-carbazoles (CZ). The quantification was confirmed by gas chromatography using triple quadrupole tandem mass spectrometry (Shimadzu GC-MS/MS-TQ8040) and gas chromatography coupled mass spectrometry (Agilent 7890A-5973 GC-MS). The developed method was validated by testing the following parameters: linearity, instrument limit of detection (LOD), instrument limit of quantification (LOQ), method limit of detection (MLD), method limit of quantification (MLQ), matrix effect (ME), accuracy, and precision. All compounds showed good linearity in the range of 0.005-0.2 µg/mL with correlation coefficients higher than 0.992. The method demonstrated satisfactory recoveries (ranging from 71.21 to 105.04%) for most of the compounds with relative standard deviation precision (RSD) < 10.46%, except 3-BCZ (Recovery = 67.53%, RSD = 2.83%). The values of LOD and LOQ varied from 0.05 to 0.24 ng and 0.14 to 0.92 ng, respectively, while those of MLD and MLQ ranged from 0.02 to 0.12 ng/g wet weight (ww) and 0.07 to 0.45 ng/g ww, respectively. The developed method represents a reliable tool for the routine analysis of PHCZ congeners in invertebrate animals.

Sediment-water distribution and potential sources of polyhalogenated carbazoles in a coastal river locating at a north metropolis, China.

The fate and transformation of PHCZs in the coastal river environment are not yet comprehensively understood. Paired river water and surface sediment were collected, and 12 PHCZs were analyzed to find out their potential sources and investigate the distribution of PHCZs between river water and sediment. The concentration of ∑PHCZs varied from 8.66 to 42.97 ng/g (mean 22.46 ng/g) in sediment and 17.91 to 81.82 ng/L (mean 39.07 ng/L) in river water. 18-B-36-CCZ was the dominant PHCZ congener in sediment, while 36-CCZ was in water. Meanwhile, the logKoc values for CZ and PHCZs were among the first calculated in the estuary and the mean logKoc varied from 4.12 for 1-B-36-CCZ to 5.63 for 3-CCZ. The logKoc values of CCZs were higher than those of BCZs, this may suggest that sediments have a higher capacity for accumulation and storage of CCZs than highly mobile environmental media.