Dihalogenated nitrophenols exposure induces developmental neurotoxicity in zebrafish embryo.

2,6-Dihalogenated nitrophenols (2,6-DHNPs) are emerging halogenated nitroaromatic pollutants that have been detected in various water environments. However, there is currently limited research available regarding their potential impacts on locomotion behavior and neurotoxicity. Therefore, this study utilized zebrafish embryos to investigate the potential neurotoxic effects of 2,6-DHNPs by examining their impact on the nervous system at a concentration defined as 10% of the median lethal concentration. Our findings demonstrated that exposure to 2,6-DHNPs resulted in a significant 30 % decrease in the total swimming distance of zebrafish larvae, accompanied by notable impairments in motor neuron development and central nervous system. These effects were evidenced by a substantial 25% decrease in axonal growth, as well as disruptions in synapse formation and neuronal differentiation. Additionally, neurotransmitter analysis revealed marked decreases of 40%, 35%, and 30% in dopamine, 5-hydroxytryptamine, and acetylcholine levels respectively, highlighting disturbances in their synthesis, transport, and degradation mechanisms. These results emphasize the considerable neurotoxicity of 2,6-DHNPs at concentrations previously considered safe; thus necessitating a re-evaluation of environmental risk assessments and regulatory standards for such emerging contaminants.

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.

The combination of multiple linear regression and adaptive neuro-fuzzy inference system can accurately predict trihalomethane levels in tap water with fewer water quality parameters.

Artificial Neural Network (ANN) models are accurate in predicting the levels of disinfection by-products (DBPs) in drinking water. However, these models are not yet practical due to the large number of parameters involved, which should take a significant amount of time and cost to detect. Developing accurate and reliable prediction models of DBPs with fewest parameters is essential in the management of drinking water safety. This study used the adaptive neuro-fuzzy inference system (ANFIS) and radial basis function artificial neural network (RBF-ANN) to predict the levels of trihalomethanes (THMs), the most abundant DBPs in drinking water. Two water quality parameters identified by multiple linear regression (MLR) models were used as model inputs, and the quality of the models was assessed based on criteria such as correlation coefficient (r), mean absolute relative error (MARE), and the percentage of predictions with absolute relative error less than 25% (NE<25%) and over than 40% (NE>40%), etc. The results showed that the ANFIS models had higher correlation coefficients (r = 0.853-0.898) and prediction accuracy (NE<25% = 91%-94%) compared to RBF-ANN models (r = 0.553-0.819; NE<25% = 77%-86%) and traditional MLR models (r = 0.389-0.619; NE<25% = 67%-77%). Conversely, the prediction error, as indicated by MARE and NE>40%, showed the opposite trend: ANFIS models (MARE = 8%-11%; NE>40% = 0-5%) < RBF-ANN models (MARE = 15%-18%; NE>40% = 5%-11%) < MLR models (MARE = 19%-21%; NE>40% = 11%-17%). The present study provided a novel approach for constructing high-quality prediction models of THMs in water supply systems using only two parameters. This method holds promise as a viable alternative for monitoring THMs concentrations in tap water, thereby contributing to the improvement of water quality management strategies.

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.

Precursor characteristics of mono-HAAs during chlorination and cytotoxicity of mono-HAAs on HEK-293T cells.

Monohaloacetic acids (mono-HAAs), a class of disinfection by-products widely occurred in drinking water, receives significant attention due to their extremely high toxicity. Many studies on the biological toxicity of mono-HAAs have been reported, yet the toxic effects of mono-HAAs on human renal cells (kidney is one of the target organs for disinfection by-products) has not been involved. Studies on organic precursors for mono-HAAs formation were also very limited due to their lower levels as compared to di-HAAs and tri-HAAs. Based on this, the formation of mono-HAAs after chlorination of some typical source water samples and their relationship with water quality parameters were investigated. Meanwhile, the cytotoxicity of monochloroacetic acid (MCAA), monobromoacetic acid (MBAA), and monoiodoacetic acid (MIAA) were tested using human embryonic kidney cells (HEK-293 T cells). The results showed that the levels of mono-HAAs formed during chlorination of source water samples were between 0.44 and 0.87 µg/L. Formation of MBAA positively (p < 0.05) correlated with bromide ion and dissolved organic carbon, but negatively (p < 0.01) correlated with SUVA254 (specific UV absorbance at 254 nm), while formation of MCAA was only positively (p < 0.05) related with SUVA254. These results suggested that although MCAA and MBAA both belong to the mono-HAAs, the characteristics of their organic precursors differ significantly. MCAA precursors have high aromaticity and are more hydrophobic, yet MBAA precursors have low aromaticity and are more hydrophilic. The half-lethal concentrations (LC50) of MCAA, MBAA, and MIAA on HEK293T cells were 1196-1211 µM, 16.07-18.96 µM, and 6.08-6.17 µM, respectively. An in-depth analysis showed that the cytotoxicity of mono-HAAs on HEK 293 T cells could not be explained by the parameters concerning cellular uptake (e.g., logP and pKa), but the SN2 reaction of C-X bond with cellular molecules (e.g., glyceraldehyde-3-phosphate dehydrogenase, etc) may be the relevant cause for the cytotoxicity of mono-HAAs on HEK 293 T cells.

Analysis of synthetic cannabinoids in wastewater of major cities in China.

Synthetic cannabinoids (SCs) could pose serious health risks to its users. It is necessary to monitor its community consumption. Wastewater-based epidemiology is a potentially useful approach in this regard. However, limited research has been conducted to investigate the occurrence of SCs in wastewater. In this study, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was optimized to analyze 8 SCs and metabolites (in total 16 analytes) in wastewater. The limit of quantification for this method for certain analytes in wastewater was as low as 0.03 ng L-1. The validated method was used to examine the stability of the analytes under different conditions and to examine their occurrence in wastewater collected from 31 major cities across China. The overwhelming majority of the analytes were stable within 24 h, even at room temperature. However, 5-fluoro MDMB-PICA and MDMB-4en-PINACA butanoic acid metabolite showed significant degradation within 120 days even when stored at -20 °C or -80 °C. At least one cannabinoid or their metabolite was detected in 21 cities. In the city with the highest detection rate, at least one synthetic cannabinoid or metabolite was detected in 95% of samples of the city. MDMB-4en-PINACA butanoic acid metabolite had the highest detection frequency (in 13.4% of the samples). These results indicated that SCs were used in a significant number of Chinese cities. A few parent drugs (MDMB-4en-PINACA, ADB-BUTINACA, 5-fluoro MDMB-PICA, 4-fluoro MDMB-BUTINACA) were detected in a small fraction of wastewater samples, possibly due to release from manufacturing of these cannabinoids or illegal addition of electronic cigarettes.

Using simple and easy water quality parameters to predict trihalomethane occurrence in tap water.

Monitoring of disinfection by-products (DBPs) in water supply system is important to ensure safety of drinking water. Yet it is a laborious job. Developing predictive DBPs models using simple and easy parameters is a promising way. Yet current models could not be well applied into practice because of the improper dataset (e.g. not from real tap water) they used or involving the parameters that are difficult to measure or require expensive instruments. In this study, four simple and easy water quality parameters (temperature, pH, UVA254 and Cl2) were used to predict trihalomethane (THMs) occurrence in tap water. Linear/log linear regression models (LRM) and radial basis function artificial neural network (RBF ANN) were adopted to develop the THMs models. 64 observations from tap water samples were used to develop and test models. Results showed that only one or two parameters entered LRMs, and their prediction ability was very limited (testing datasets: N25 = 46-69%, rp = 0.334-0.459). Different from LRM, the prediction accuracy of RBF ANNs developed with pH, temperature, UVA254 and Cl2 can be improved continuously by tweaking the maximum number of neuron (MN) and Gaussian function spread (S) until it reached best. The optimum RBF ANNs of T-THMs, TCM and BDCM were obtained when setting MN = 20, S = 100, 100.1 and 60, respectively, where the N25 and rp values for testing datasets reached 85-92% and 0.813-0.886, respectively. Accurate predictions of THMs by RBF ANNs with these four simple and easy parameters paved an economic and convenient way for THMs monitoring in real water supply system.

Environmentally relevant concentrations of arsenic induces apoptosis in the early life stage of zebrafish.

Arsenic (As) in the aquatic environment is a considerable environmental issue, previous studies have reported the toxic effects of low concentrations (≤ 150 µg/L) of As on fish. However, limited information is available regarding the impact of low levels of As on apoptosis. To evaluate this, zebrafish embryos were exposed to different concentrations (0, 25, 50, 75, and 150 µg/L) of As (arsenite [AsIII] and arsenate [AsV]) for 120 h. Our results indicated that low concentrations of AsIII exposure significantly inhibited the survival of zebrafish larvae, and significantly increased the transcription of Caspase-9 and Caspase-3, the ratio of Bax/Bcl-2 transcription, and protein levels of Caspase-3. In contrast, AsV decreased the ratios of Bax/Bcl-2 transcription and protein levels, as well as protein levels of Caspase-3. Our data demonstrated that AsIII and AsV exert different toxic effects, AsIII induced apoptosis via the mitochondrial pathway and the extrinsic pathway, while AsV induced apoptosis only via the mitochondrial pathway.

Analysing wastewater to estimate fentanyl and tramadol use in major Chinese cities.

The misuse of fentanyl and more recently tramadol in the population has caused an opioid crisis in several countries and drawn much public attention worldwide. However, there is a gap of information on the potential misuse of fentanyl and tramadol in China. This study aims to fill this gap by analysing fentanyl and tramadol in wastewater of major cities across China to estimate their use. Wastewater samples were collected from 30 cities located across all seven geographic regions of China, from 2016 to 2019. Fentanyl was detected in only a few samples, suggesting a low prevalence of this potent opioid drug in China. Meanwhile, tramadol was found in most samples with concentrations ranging up to 186 ng/L. The per capita daily consumption of tramadol estimated from wastewater across China ranged from 6 mg/d/1000 in. to 213 mg/d/1000 inh. The consumption of tramadol seems to be similar among all the days of the week. Tramadol use is overall higher in Northeast China than in other regions, which is different from heroin, another popular opioid in China. Temporally, there is a significant decrease in tramadol use in major cities of China from 2016 to 2019. The results of our study suggested that tramadol use in China was predominantly from pharmaceutical prescription and not as prevalent as in other countries.

Assessment on the adverse effects on different kinds of fish induced by methamphetamine during the natural attenuation process based on adverse outcome pathway.

The adverse effects on model fish induced by methamphetamine (METH) have been revealed. However, the toxicity of METH on different kinds of non-model fish during the natural attenuation remained unclear. Hence, in this study, we for the first time established a static lab-scale aquatic ecosystem spiked with METH (initial levels at 25 µg/L) for 40 days to estimate its metabolism and toxicity in Chinese medaka, rosy bitterling, loach, and mosquito fish. The concentrations of METH in water and fish's brain were detected termly. The physiological functions, histopathology of brain, neurotransmitters contents, and expressions of associated genes of the four kinds of fish were determined at day 0, 20, and 40, respectively. The results indicated METH could be remarkably accumulated in fish brains with the distribution factor vs water (DFw) at 232.5-folds, and attenuated both in water and fish body during the exposure. METH caused physiological functions (i.e., swimming trajectories, locomotion distances, and feeding rates) disorders of the four kinds of fish, and stimulated surfacing behavior of loach. Tissue and macro/micromolecular biomarkers including histopathology, neurotransmitters (i.e., dopamine, serotonin, and norepinephrine), and mRNA, were similarly affected by METH. Mitogen-activated protein kinase (MAPKs) signaling pathway, P53-regulated apoptosis signaling pathway, N-methyl-d-aspartate-dopamine system, and mTOR signaling pathway of different kinds of fish were regulated by METH. Additionally, the impairments of the physiological and macromolecular indicators of fish could be alleviated as the natural attenuation of METH occurred. All the biomarkers, as well as the recovery effects during the exposure were integrated onto an adverse outcome pathway (AOP) framework. The key event was the micromolecular indicators (genes). The adverse outcomes at individual and population levels would result in the ecological consequences, implying the imperative to consider the natural attenuation process while assessing the environmental risk of METH.

Cyclophosphamide induced physiological and biochemical changes in mice with an emphasis on sensitivity analysis.

The widespread use of cyclophosphamide (CP) in medical treatment had caused ubiquitous contamination in the environment. To data, many studies have been carried out on the toxic effect of CP. However, among these toxic effects of CP, which are the most sensitive remains unclear. Present study aimed to investigate the toxicity of CP on mice and evaluate the sensitivity of physiological-biochemical parameters upon exposure of mice to CP. Results showed that as compared with the control group, CP caused significant reduction in body weight (p < 0.01), spleen coefficient (p < 0.01), leukocyte density (p < 0.01) and alanine transaminase (ALT) in kidney (p < 0.01); However superoxide dismutase (SOD), malondialdehyde (MDA), ALT in liver and creatinine (Cr) in kidney significantly (p < 0.05) increased. Among the suppressed physiological and biochemical parameters, the sensitivity to CP toxicity was generally ranked as body weight > leukocyte density > ALT in kidney > spleen coefficient; while among the stimulated parameters, the sensitivity was ranked as MDA (liver) > Cr (kidney) > ALT (liver). Overall, the most sensitive parameters to CP toxicity may be associated with growth, immune system and the normal function of liver and kidney.

Impact of ketamine on the behavior and immune system of adult medaka (Oryzias latipes) at environmentally relevant concentrations and eco-risk assessment in surface water.

This work for the first time investigated the bioconcentration factor (BCF), toxicity, and eco-risk of KET using adult medaka fish (Oryzias latipes) as model organism after exposure at environmental concentrations (0.05-0.5 µg L-1) and higher levels (5-100 µg L-1) for 90 days. The BCF of KET was approximately 1.07- to 10.94- folds. The behavioral functions, including swimming properties, feeding rate, and food preference, were significantly impacted by KET (≥0.05 µg L-1). After 90-days exposure, KET induced histological abnormalities in liver and kidney tissue at 0.1 and 0.2 µg L-1, respectively. Additionally, the condition factor, hepatic-somatic index (HSI), and nephric-somatic index (NSI) of medaka were markedly impacted by KET treatment at 0.5, 0.5, and 0.1 µg L-1, respectively. Morphological inflammation (i.e., haemorrhage and erosion) in the fish body was observed exposed to KET, and the EC10 value was 0.407 µg L-1. Alterations in the expressions of genes (i.e., cacna1c, oxtr, erk1, and c-fos) and proteins (i.e., OXT and PKA), involved in in calcium ion channels induced by KET, could partly elucidate the underlying mechanism of the toxicity. The inflammatory risk to fish posed by KET in some rivers in southern China was at high level, suggesting the long-term concentration monitoring was required.

[Occurrence of Tramadol and Fentanyl Use in Domestic Wastewater in Beijing].

Tramadol (TRA) and fentanyl (FEN) are used as common painkillers in clinical practice, but they have been increasingly abused in recent years due to their addictive nature. Two substances and their metabolites enter wastewater through urine and are collected and treated by wastewater treatment plants (WWTPs) before being discharged into the aquatic environment. In this study, wastewater analysis was performed to examine the patterns of TRA and FEN use in the urban area of Beijing for the first time. Influent and effluent samples were collected from 23 WWTPs during two sampling campaigns. Concentrations TRA in influents were found to range from(10.2±8.7)to(175.3±59.7) ng·L-1, while FEN was not detected in most of the samples, or occurred at very low concentrations. Relatively low TRA removal was observed at plants with activated sludge processes. Moreover, TRA loads in the central area of Beijing were significantly higher than those in the suburban areas. Annual TRA use was estimated through wastewater-based epidemiology. The greatest TRA use, approximately 202.5 kg, was found in Haidian district. Seasonal variation in TRA loads was significant, with greater use in the summer than in winter. The method presented in this study can be used as an important reference for monitoring TRA and FEN use via wastewater-based epidemiology and for assessing the risk of the abuse of these compounds in China.

Monitoring consumption of methadone and heroin in major Chinese cities by wastewater-based epidemiology.

BACKGROUND: Methadone maintenance treatment (MMT) services have been used in China for treatment of heroin dependence. But no study has been conducted to assess the appropriateness of MMT distribution and the potential abuse of methadone in China. This study aims to do that through a nationwide estimation of methadone consumption in China via wastewater-based epidemiology and subsequently compare it with MMT data and level of heroin abuse. METHODS: Wastewater samples were collected from 53 wastewater treatment plants in 27 major cities that cover all geographic regions of China. Methadone and pure heroin consumptions were estimated based on influent concentrations of methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), morphine and codeine. RESULTS: Drug residues were detected in most samples. The ratio of EDDP/methadone was around 2 in influents and methadone and EDDP loads were strongly correlated, indicating that they originated from human consumption. Both influent methadone and EDDP loads in Southwest and Northwest China were significantly higher than those in other regions. The highest estimated consumptions of methadone and heroin in China were 22.0 ±â€¯2.1 mg/1000 in./d and 263.9 ±â€¯115.9 mg/1000 in./d, respectively. There was a significant positive correlation between methadone and heroin consumptions. CONCLUSIONS: Consumption of methadone in China was primarily from MMT services. The use of methadone and heroin displayed a clear geographical pattern: it is higher in the western inland regions than in the eastern regions. This study has shown that the distribution of MMT services is reflective of the level of heroin abuse in different regions of China.

Impacts of methamphetamine and ketamine on C.elegans's physiological functions at environmentally relevant concentrations and eco-risk assessment in surface waters.

In this work, C. elegans as a model organism was treated with methamphetamine (METH) and ketamine (KET) to assess its eco-toxicity at a range (0.05-250 µg L-1) that covers environmentally relevant concentrations (0.05-0.5 µg L-1). METH (≥0.05 µg L-1) and KET (≥0.5 µg L-1) significantly affected the feeding rate, locomotion, gustation and olfaction (P < 0.05), which may result in pronounced disturbance to aquatic ecology. Alterations in the contents of neurotransmitters (i.e., octopamine (OA), dopamine (DA), and serotonin (5-HT)) correlated with the physiology change. The metabolic activities and the antioxidase activity (i.e., superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)) of METH and KET in C. elegans were different, which could partly explain the difference of the physiological changes induced by the two substances. Moreover, these two drugs could induce vulva deformity, and the 50% effect concentrations were 620.34 µg L-1 for METH and 54.39 µg L-1 for KET, respectively. The risk quotients (RQ) in two Chinese rivers, the Shenzhen and Liangshui River, were calculated to assess eco-risks of METH and KET. RQs of KET in the Shenzhen River were over 0.1 at the medium risk level, indicating that eco-risks of illicit drugs to aquatic organism cannot be overlooked.

Estimating population exposure to phthalate esters in major Chinese cities through wastewater-based epidemiology.

Phthalate esters (PAEs) are widely used in consumer and industrial products and may thus pose significant health risks. Urine analysis, which has usually been applied to assess the health risks of PAEs, has the drawback of small sample sizes and insufficient representativeness. Wastewater-based epidemiology (WBE) collects wastewater samples containing urine from the entire community and thus is more representative than urine samples. In this work, exposure levels and health risks of PAEs were estimated on a national scale for the first time through the WBE approach. Wastewater samples were collected from 54 wastewater treatment plants in 27 major cities that cover all of the geographic regions of China. The estimated ∑5PAEs exposure levels ranged from 290 µg/inh/d (Lhasa) to 3642 ±â€¯467 µg/inh/d (Zhengzhou) with a mean level of 2184 ±â€¯1173 µg/inh/d. Di-n-butyl phthalate accounted for the highest proportion (65%) in the total exposure level. The ∑5PAEs exposure levels in Southwest China were significantly lower than those in other regions due to the low production and consumption of plastics in the region. The health risks of PAEs were assessed by comparing the estimated daily exposure levels to the acceptable daily exposure levels. For adults, the hazard index that represents cumulative risk of PAEs was above or below 1 depending on a particular reference dose (total daily intake values or the reference dose for anti-androgenicity) that was used for risk calculation. In contrast, the hazard index was above 1 for a significant number of cities for children, regardless of which reference dose was used. The results indicate that health risks of PAE exposure in China cannot be overlooked. Children in China are under considerably greater risks than adults, which warrants further research or proper regulation of PAE use in China.

Biodegradation of methamphetamine and ketamine in aquatic ecosystem and associated shift in bacterial community.

Methamphetamine (METH) and ketamine (KET) are widely detected in surface waters and thus may pose threat to aquatic organisms. However, their degradation in aquatic systems and the effects on bacterial community were unknown. The present study investigated the biodegradation process of METH and KET in river waters and sediments. Three microcosms were examined over 40-days' incubation under (i) aerobic and illumination conditions, (ii) anaerobic condition exposed to light, (iii) anaerobic-dark condition. Statistically significant biodegradation of METH and KET (1 mg L-1) was observed in all treatments. The half-lives under the examined conditions indicate that the two drugs were refractory in aquatic environment. Moreover, there were no pronounced absorption and photolysis observed in this work. Illumina MiSeq sequencing analysis revealed that Methylophilaceae, Saprospiraceae, WCHB1-69, Desulfobulbaceae, Porphyromonadaceae, FamilyXI, Peptococcaceae, and Rhizobiaceae were the predominant candidatus families during KET and METH biodegradation, and the preponderance would impair other microorganisms' prosperity since them were scarcely detected in the wild. Meanwhile, canonical correlation analysis (CCA) indicates that METH as an environmental factor may affect bacterial community structure in field water samples.

[Optimization and Validation of the Analytical Method to Detect New Psychoactive Substances in Wastewater].

New psychoactive substances (NPS) are emerging continuously, as the crackdown against traditional drugs becomes stricter. Metabolites of NPSs and the unchanged drugs enter wastewater through urine and are collected and treated by wastewater treatment plants before being discharged into the aquatic environment. Based on previous research, solid phase extraction (SPE) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) were used and the detection method of 11 NPSs was optimized. Influences of wastewater pretreatment procedures, including SPE columns; pH values of water samples; and processes of flushing and redissolving on NPS recoveries were compared. It was shown that the best recovery was achieved when Oasis MCX columns were used and the pH value of wastewater samples was adjusted to 2. A flushing process with 2 mL water at pH=2 followed by 2 mL methanol was needed. Redissolution of the residue after evaporation was best achieved with 400 µL of a 20% methanol water solution. It was indicated by retention times, recoveries, matrix effects, limits of detection, limits of quantification, as well as precision that C18-UPLC-MS/MS and the optimized method are efficient and valid. Influent and effluent samples from 11 municipal wastewater treatment plants in Beijing were analyzed using the optimized method for validation. The validated method can be used as an important reference for monitoring NPSs via wastewater-based epidemiology and for assessing the risk of NPS abuse in China.

Enantiomeric profiling of amphetamine and methamphetamine in wastewater: A 7-year study in regional and urban Queensland, Australia.

Enantiomeric profiling was used in this study to investigate the consumption of amphetamine and methamphetamine in regional and urban Southeast Queensland, Australia over a period of seven years. S(+) methamphetamine was predominantly consumed in both urban and regional areas, showing a two and three fold increase in urban and regional catchments respectively between 2011 and 2017. The ratio of amphetamine to methamphetamine (AMP/METH) in wastewater reflected the expected excretion profile of methamphetamine consumption indicating the presence of amphetamine in this study was primarily the result of methamphetamine metabolism. However, the occasional occurrence of R(-) amphetamine in samples containing higher AMP/METH ratios, suggested the consumption of racemic amphetamine. The R(-) methamphetamine enantiomer was also identified in several samples, possibly indicative that the phenyl-2-propanone (P2P) synthesis process rather than the more typical reduction of ephedrines was also being used to manufacture methamphetamine. Furthermore, we identified two samples with a significantly different enantiomer ratio for the METH and AMP as well as a much lower AMP/METH concentration ratio suggesting contribution from direct disposal of methamphetamine into the sewer. This study demonstrated that enantiomeric profiling in wastewater-based epidemiology can provide valuable information for evaluating the origin of amphetamine in wastewater as either a metabolite of methamphetamine consumption or amphetamine itself.

Estimating heroin abuse in major Chinese cities through wastewater-based epidemiology.

Heroin consumption in major cities across China was estimated for the first time via wastewater-based epidemiology. Influent and effluent wastewater samples were collected from 49 wastewater treatment plants (WWTPs) in 24 major cities that cover all the geographic regions of the country. Concentrations of morphine, 6-acetylmorphine, and codeine were measured. Near complete removal of morphine by wastewater treatment processes was observed, whereas removal rates of codeine were slightly lower. Morphine loads were much higher than codeine loads at most WWTPs in China, a trend opposite to that in many European countries. In addition, morphine and codeine loads were strongly correlated at most WWTPs, indicating morphine and codeine in wastewater were predominantly from the same source, street heroin. At WWTPs in Guangzhou and Shenzhen, codeine loads were considerably higher than morphine loads, consistent with previous reports of codeine abuse (e.g., as cough syrup) among middle and high school students in Guangdong province. Heroin consumption was derived based on morphine loads and taking into account therapeutic use of morphine and codeine, as well as contribution of codeine and acetylcodeine in street heroin. Highest heroin consumption was observed in northwestern and southwestern China. The average heroin consumption of the sampled cities was 64.6±78.7mg/1000inh/d. The nation-wide average heroin consumption was much lower than that of methamphetamine, consistent with seizure data and numbers of registered heroin and methamphetamine users in China.