Metabolites In Wastewater Research Articles

International Workers' Day: Consumption Patterns of Morphine, Codeine, and Methamphetamine in Urban and Rural Areas Based on Wastewater-Based Epidemiology.

Wastewater-based epidemiology (WBE) is a reliable means to estimate drug consumption in a specific population. By measuring the concentration of drug residues or metabolites in wastewater, the consumption behavior pattern of a specific population can be deduced. Using the WBE method, the present study, for the first time, continuously monitored the differences in the consumption of morphine (MOR), codeine (CODE), and methamphetamine (METH) in three wastewater-treatment plants in a city and two surrounding villages in Xinjiang, China during International Workers' Day and the following week. The wastewater samples were pretreated by solid-phase extraction and then analyzed by high-performance liquid chromatography-tandem mass spectrometry. Methamphetamine was not detected in rural areas and was detected only on International Workers' Day in urban areas. According to the estimation of per capita consumption, the per capita consumption of MOR, CODE, and METH in urban inhabitants was 12.04 to 23.39, 10.44 to 16.39, and 1.31 mg/day/1000 inhabitants. The per capita consumption of MOR and CODE in rural areas was 5.19 to 8.35 and 2.56 to 3.52 mg/day/1000 inhabitants. The consumption of MOR in urban and rural areas was significantly higher than that of CODE and METH. During International Workers' Day, workdays, and weekends, the consumption of MOR and CODE in urban areas is significantly higher than that in rural areas. Compared with those on weekends, the consumption of urban MOR and CODE increased more during International Workers' Day. The consumption of MOR in urban areas showed a weekend effect. The present study can provide information for subsequent research and government departments. Environ Toxicol Chem 2024;00:1-9. © 2024 SETAC.

Validation of LC-MS/MS method for opioid monitoring in Valencia City wastewater: Assessment of synthetic wastewater as potential aid

In this study, a comprehensive and sensitive method for the simultaneous detection of 17 opioids (OPs) and their human metabolites in wastewater using high-performance liquid chromatography coupled to tandem mass spectrometry was validated. The chromatographic separations of opioids were carried out on a Kinetex® Biphenyl column (1.7 μm, 100 Å, 50 × 2.1 mm). A synthetic wastewater approach was used for recovery studies to mimic a contaminant-free matrix. Two solid-phase extraction (SPE) sorbents (hydrophilic-lipophilic balance and mixed mode with the previous phase and a weak cationic exchange) were studied to optimize sample treatment and obtain higher recoveries. The mixed mode was chosen because the recoveries of 17 target analytes at three spiked concentrations (25, 50, and 100 ng mL−1) were > 80 % for 75 % of the analytes in a simulated wastewater. The intra- and inter-day relative standard deviations (RSDs) were between ±1 % and ±20 %. The method limits of quantification ranged from 5 to 25 ng L−1, the only exceptions being heroin (275 ng L−1) and morphine-3β-glucuronide (250 ng L−1). Suppression/enhancement is comparable between the synthetic and the influent wastewater. The analytical method was applied to the OPs analysis in twenty-one influent samples collected from the treatment plants treating the wastewater of Valencia City (Spain). Twelve OPs were detected with total daily concentrations ranging from 1 ng L−1 to 2135 ng L−1. The widespread presence of these compounds in water suggests potential widespread exposure, highlighting the need for increased environmental awareness. Furthermore, the estimated daily intake results raise concerns about opioid use as a potential future health and social issue.

Wastewater-based epidemiology methodology to investigate human exposure to bisphenol A, bisphenol F and bisphenol S.

Wastewater-based epidemiology (WBE) has become an invaluable tool for tracking the evolution of use or exposure of/to numerous substances. Bisphenols, commonly utilized in manufacturing plastic goods, have been categorized as endocrine disrupting chemicals, underscoring the critical need for real-time data on their local-level exposure to safeguard public health. In this study, we have developed a novel analytical method and WBE framework for the assessment of population-level exposure to bisphenol A (BPA) and its most prominent substitutes, bisphenols F and S (BPF and BPS), through the determination their Phase II metabolites in wastewater by WBE. Stability and exclusivity tests denoted that glucuronides are not stable in sewage, whereas sulfate metabolites are good biomarkers. Therefore, a solid-phase extraction followed by liquid chromatography-tandem mass spectrometry method was developed for the bisphenols’ monosulfates and BPA bissulfate. The analytical method was validated with three different wastewater matrices, providing trueness (as recovery) in the 79–112 % range with relative standard deviations < 12 %, and method quantification limits below 2 ng L−1 for monosulfates, but higher (35 ng L−1) for BPA bissulfate. Subsequently, the method was applied to 24h-composite raw wastewater samples collected over a week in 4 different locations in Spain and Portugal. BPA bissulfate was not detected, but the three monosulfate metabolites of each bisphenol were positively detected in the samples, being the metabolite of BPA the most prevalent, followed by those of BPF and BPS. Community-wide BPA intake was then estimated to be higher than the European Food Safety Agency (EFSA) tolerable daily intake (TDI) of 2 × 10−4 µg kg−1day−1 in all locations. In the case of BPF and BPS, there is not enough metabolism data or even established limit, but they would also surpass safe levels in several locations if a similar metabolism and TDI would be assumed. This innovative method could be used to a larger set of wastewater-treatment plants as an early-warning approach on human exposure to bisphenols.

Application of Magnetic Materials Combined with Echo® Mass Spectrometry System in Analysis of Illegal Drugs in Sewage.

The aim of this study is to solve the problems of the complicated pretreatment and high analytical cost in the detection technology of trace drugs and their metabolites in municipal wastewater. A high-performance magnetic sorbent was fsynthesized for the enrichment of trace drugs and their metabolites in wastewater to develop a magnetic solid-phase extraction pretreatment combined with the acoustic ejection mass spectrometry (AEMS) analytical method. The magnetic nanospheres were successfully prepared by magnetic nanoparticles modified with divinylbenzene and vinylpyrrolidone. The results showed that the linear dynamic range of 17 drugs was 1-500 ng/mL, the recovery was 44-100%, the matrix effect was more than 51%, the quantification limit was 1-2 ng/mL, and the MS measurement was fast. It can be seen that the developed magnetic solid-phase extraction (MSPE) method is a good solution to the problems of the complicated pretreatment and analytical cost in the analysis of drugs in wastewater. The developed magnetic material and acoustic excitation pretreatment coupled with mass spectrometry analysis method can realize the low-cost, efficient enrichment, and fast analysis of different kinds of drug molecules in urban sewage.

Development and application of a sustainable approach for the determination of 95 pharmaceutical substances and metabolites in urban wastewater by means of ultra-high-performance liquid-chromatography-tandem mass spectrometry

ABSTRACT In the last few years, the interest in pharmaceutical drugs and their metabolites as environmental pollutants is gaining growing importance. Regular monitoring and timely actions are decisive to implement appropriate water resource management strategies and to evaluate the efficiency of traditional or innovative wastewater treatment plants (WWTPs). This study describes the development and validation of an analytical procedure using 10 mL sample volume followed by direct-injection in ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of 95 pharmaceutical drugs and 10 of their main metabolites in wastewater. Adequate sensitivity was recorded for all target analytes, with limits of detection below 5 ng/L for 60 out of 95 analytes and recoveries exceeding 80% for all the analytes under study. A total of 42 target analytes were detected in almost all sites, and limited differences were observed among several pharmaceutical drug arrays found at different sampling sites. In addition, an abatement yield higher than 50% was observed for only 12 of the 42 detected substances. The procedure, which combined direct injection and small sample volume collection, showed great potential and efficiency for the high-performance determination of pharmaceutical drugs in wastewater.

Monitoring of over-the-counter (OTC) and COVID-19 treatment drugs complement wastewater surveillance of SARS-CoV-2

BackgroundThe application of wastewater-based epidemiology to track the outbreak and prevalence of coronavirus disease (COVID-19) in communities has been tested and validated by several researchers across the globe. However, the RNA-based surveillance has its inherent limitations and uncertainties.ObjectiveThis study aims to complement the ongoing wastewater surveillance efforts by analyzing other chemical biomarkers in wastewater to help assess community response (hospitalization and treatment) during the pandemic (2020–2021).MethodsWastewater samples (n = 183) were collected from the largest wastewater treatment facility in Suffolk County, NY, USA and analyzed for COVID-19 treatment drugs (remdesivir, chloroquine, and hydroxychloroquine (HCQ)) and their human metabolites. We additionally monitored 26 pharmaceuticals including common over-the-counter (OTC) drugs. Lastly, we developed a Bayesian model that uses viral RNA, COVID-19 treatment drugs, and pharmaceuticals data to predict the confirmed COVID-19 cases within the catchment area.ResultsThe viral RNA levels in wastewater tracked the actual COVID-19 case numbers well as expected. COVID-19 treatment drugs were detected with varying frequency (9–100%) partly due to their instability in wastewater. We observed a significant correlation (R = 0.30, p < 0.01) between the SARS-CoV-2 genes and desethylhydroxychloroquine (DHCQ, metabolite of HCQ). Remdesivir levels peaked immediately after the Emergency Use Authorization approved by the FDA. Although, 13 out of 26 pharmaceuticals assessed were consistently detected (DF = 100%, n = 111), only acetaminophen was significantly correlated with viral loads, especially when the Omicron variant was dominant. The Bayesian models were capable of reproducing the temporal trend of the confirmed cases.ImpactIn this study, for the first time, we measured COVID-19 treatment and pharmaceutical drugs and their metabolites in wastewater to complement ongoing COVID-19 viral RNA surveillance efforts. Our results highlighted that, although the COVID-19 treatment drugs were not very stable in wastewater, their detection matched with usage trends in the community. Acetaminophen, an OTC drug, was significantly correlated with viral loads and confirmed cases, especially when the Omicron variant was dominant. A Bayesian model was developed which could predict COVID-19 cases more accurately when incorporating other drugs data along with viral RNA levels in wastewater.

Statistical Analysis of Methotrexate Degradation by UV-C Photolysis and UV-C/TiO2 Photocatalysis.

Methotrexate (MTX) is a folic acid analog and has been used to treat a wide variety of malignant and non-malignant diseases. The wide use of these substances has led to the continuous discharge of the parent compound and its metabolites in wastewater. In conventional wastewater treatment plants, the removal or degradation of drugs is not complete. In order to study the MTX degradation by photolysis and photocatalysis processes, two reactors were used with TiO2 as a catalyst and UV-C lamps as a radiation source. H2O2 addition was also studied (absence and 3 mM/L), and different initial pHs (3.5, 7, and 9.5) were tested to define the best degradation parameters. Results were analyzed by means of ANOVA and the Tukey test. Results show that photolysis in acidic conditions with 3 mM of H2O2 added is the best condition for MTX degradation in these reactors, with a kinetic constant of 0.028 min-1. According to the ANOVA test, all considered factors (process, pH, H2O2 addition, and experimentation time) caused statistically significant differences in the MTX degradation results.

Community infectious disease treatment with antimicrobial agents – A longitudinal one year study of antimicrobials in two cities via wastewater-based epidemiology

Antimicrobial resistance (AMR) is one of the most significant global health threats. Inappropriate and over-usage of antimicrobial agents (AAs) is a major driver for AMR. Wastewater-based epidemiology (WBE) is a promising tool for monitoring AA usage in communities which is, for the first time, explored in this large scale, longitudinal study. Two contrasting urban catchment areas have been investigated: one city and one small town in the Southwest of the UK over a 13-month period in 2018–2019. Per capita daily intake of 17 AAs and metabolites has been estimated and obtained estimates were triangulated with catchment specific AA prescription data to understand AA usage patterns (both seasons driven prescription and AA prescription compliance). Results have demonstrated positive correlations for all quantifiable parent AAs and metabolites in wastewater, and spatial variability in AA usage was observed even in neighbouring urban areas. WBE and catchment specific prescription data showed similar seasonal trends but with low correlation in intake. The reasons might be variable prescribing patterns, prescription/intake outside the studied catchment, and/or lack of patient compliance. WBE proved useful in differentiating between consumption vs topical usage and/or direct disposal of unused AA. WBE is considered superior to prescription data as it provides information on AAs prescribed outside of the monitoring catchment, e.g. HIV antivirals and TB drugs. However, data triangulation, of both prescription data and wastewater data, provides the most comprehensive approach to understanding AA usage in communities.

Determination of the pyrethroid inseticide metabolite 3-phenoxybenzoic acid in wastewater using polar organic integrative samplers and LC-MS/MS analysis

Wastewater-based epidemiology (WBE) is a methodological approach that aims to obtain epidemiological information through wastewater analysis, by identifying endogenous metabolism products. Pyrethroid insecticides are commonly used but just a few studies evaluated concentrations of pyrethroid metabolites in wastewater. A cost-effective and efficient sampling strategy for WBE studies is the use of polar organic chemical integrative samplers (POCIS). This study aimed to develop and evaluate a method for estimating the concentration of the pyrethroid insecticide 3-phenoxybenzoic acid (3-PBA) using POCIS sampling and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The sampler sorbent was extracted with methanol, followed by a double solid phase extraction (SPE) clean-up. The extraction procedure had yields in the range of 95.7–100.4 %. There were 27 collection cycles, with POCIS samplers changed every 14 days. The analyte 3-PBA was measured in all collections. The concentration of 3-PBA in the POCIS sorbent was 208.6–2456.9 ng g−1. The sampling rate was 0.096 L/day and the resulting estimated concentration in wastewater was 24.3 to 335.2 ng/L. This is the first study evaluating POCIS sampling to measure a pyrethroid metabolite in water or wastewater samples.

Identifying antimicrobials and their metabolites in wastewater and surface water with effect-directed analysis

This study aimed to identify antimicrobial contaminants in the aquatic environment with effect-directed analysis. Wastewater influent, effluent, and surface water (up- and downstream of the discharge location) were sampled at two study sites. The samples were enriched, subjected to high-resolution fractionation, and the resulting 80 fractions were tested in an antibiotics bioassay. The resulting bioactive fractions guided the suspect and nontargeted identification strategy in the high-resolution mass spectrometry data that was recorded in parallel. Chemical features were annotated with reference databases, assessed on annotation quality, and assigned identification confidence levels. To identify antibiotic metabolites, Phase I metabolites were predicted in silico for over 500 antibiotics and included as a suspect list. Predicted retention times and fragmentation patterns reduced the number of annotations to consider for confirmation testing. Overall, the bioactivity of three fractions could be explained by the identified antibiotics (clarithromycin and azithromycin) and an antibiotic metabolite (14-OH(R) clarithromycin), explaining 78% of the bioactivity measured at one study site. The applied identification strategy successfully identified antibiotic metabolites in the aquatic environment, emphasizing the need to include the toxic effects of bioactive metabolites in environmental risk assessments.

Stability of human stress hormones and stress hormone metabolites in wastewater under oxic and anoxic conditions.

Levels in wastewater of human stress biomarkers, such as cortisone (E), cortisol (F), tetrahydrocortisone (THE), and tetrahydrocortisol (THF) may serve as indicators of population wellbeing and overall health. This study examined the stability of these biosignature compounds in wastewater to inform on their applicability for use in wastewater-based epidemiology (WBE). Wastewater from two undisclosed U.S. municipalities were fortified with the above four biomarkers of stress to a concentration of 10 ppb, and their decay was studied at three temperatures (15, 25, and 35 °C) over 24 h in oxic and anoxic conditions. Samples were analyzed using liquid chromatography tandem mass spectrometry (LC-MS/MS) in conjunction with the isotope dilution method for absolute quantitation. Results demonstrated short-term persistence (24 h) of biomarkers at low temperatures (15 °C), and accelerating kinetics of decay that were positively correlated with temperature increases. Among the four biomarkers evaluated, the tetrahydro derivatives were the most long-lived sewage-borne stress biomarkers and these are recommended as prime analytical targets for use in WBE when tracking population stress. Statistical analyses using a non-parametric Wilcoxon test further revealed no significant differences (p > 0.05) between oxic and anoxic decay rates for all stress biomarkers in wastewater from all study locations, regardless of the prevailing temperature regime. This negative finding is worthy of reporting because it suggests the feasibility of straightforward modeling of stress hormone decay, irrespective of whether the sewerage system monitored contains fully filled, pressurized pipes or partially filled gravity flow pipes, whose filling level, and with it its redox conditions, are known to fluctuate over time with water use and storm events.

Automatised on-line SPE-chiral LC-MS/MS method for the enantiomeric determination of main fluoroquinolones and their metabolites in environmental water samples

Fluoroquinolones are antibiotics of significant environmental concern their extended use not only in human medicine but also in veterinary medicine and not only as therapeutic agents but also to promote livestock growth and in aquaculture. Some fluoroquinolones and their metabolites are chiral compounds. Therefore, for a proper environmental risk assessment, enantioselective analytical methods are required. In this work, an analytical method has been developed and validated for the first time automatised enantioselective determination of environmental significant fluoroquinolones and their metabolites in wastewater and surface water samples. Target fluoroquinolones were selected by considering their extended use in human (ciprofloxacin and ofloxacin) and veterinary (flumequine) medicine. The analytical method was based on on-line solid-phase extraction-chiral liquid chromatography-tandem mass spectrometry. Analysis, including sample extraction and chiral LC-MS/MS determination, was carried out in just 14 min. The method was validated for its application to surface water and effluent and influent wastewater. Accuracy values were in the range from 61.4 to 122 % in wastewater and from 73.4 to 119 % in surface water. Precision, expressed as relative standard deviation, was lower than 13.6 % for all the compounds and sample matrices. Method quantification limits were in the range from 0.2 to 50 ng/L for all the compounds in wastewater and surface water. Method application to wastewater and surface water samples revealed the enantioselective transformation of LEV into (R)-OFL in surface water and the prevalence of OH-FLU D2 with respect to OH-FLU D1 in influent wastewater.

Direct injection ultra-performance liquid chromatography-tandem mass spectrometry for the high-throughput determination of 11 illicit drugs and metabolites in wastewater

The measurement of illicit drugs in wastewater is increasingly being adopted as a method for objective monitoring of population-level illicit drug use. This work describes the first small-volume direct-injection ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of 11 illicit drugs and metabolites in wastewater. The method required an injection volume of only 30 µL of wastewater sample, the limits of detection (LOD) ranged from 0.4 ng/L to 2 ng/L and the lower limits of quantitation (LLOQ) ranged from 1 ng/L to 5 ng/L. Application of the method to real wastewater samples collected from wastewater treatment plants revealed morphine in all samples, together with other illicit drugs (methamphetamine, codeine, ketamine, and nor-ketamine) in some samples. Small-volume direct injection showed great potential as an efficient method for the high-throughput determination of illicit drugs in wastewater.

Population Health Metrics During the Early Stages of the COVID-19 Pandemic: Correlative Pilot Study.

BackgroundCOVID-19 has caused nearly 1 million deaths in the United States, not to mention job losses, business and school closures, stay-at-home orders, and mask mandates. Many people have suffered increased anxiety and depression since the pandemic began. Not only have mental health symptoms become more prevalent, but alcohol consumption has also increased during this time. Helplines offer important insight into both physical and mental wellness of a population by offering immediate, anonymous, cheap, and accessible resources for health and substance use disorders (SUD) that was unobstructed by many of the mandates of the pandemic. Further, the pandemic also launched the use of wastewater surveillance, which has the potential for tracking not only population infections but also consumption of substances such as alcohol.ObjectiveThis study assessed the feasibility of using multiple public surveillance metrics, such as helpline calls, COVID-19 cases, and alcohol metabolites in wastewater, to better understand the need for interventions or public health programs in the time of a public health emergency.MethodsEthanol metabolites were analyzed from wastewater collected twice weekly from September 29 to December 4, 2020, in a Midwestern state. Calls made to the helpline regarding housing, health care, and mental health/SUD were correlated with ethanol metabolites analyzed from wastewater samples, as well as the number of COVID-19 cases during the sampling period.ResultsCorrelations were observed between COVID-19 cases and helpline calls regarding housing and health care needs. No correlation was observed between the number of COVID-19 cases and mental health/SUD calls. COVID-19 cases on Tuesdays were correlated with the alcohol metabolite ethyl glucuronide (EtG). Finally, EtG levels were negatively associated with mental health/SUD helpline calls.ConclusionsAlthough helpline calls provided critical services for health care and housing-related concerns early in the pandemic, evidence suggests helpline calls for mental health/SUD-related concerns were unrelated to COVID-19 metrics. Instead, COVID metrics were associated with alcohol metabolites in wastewater. Although this research was formative, with continued and expanded monitoring of population metrics, such as helpline usage, COVID-19 metrics, and wastewater, strategies can be implemented to create precision programs to address the needs of the population.

Regional changes in stimulant drug use during COVID-19 pandemic in Finland: A wastewater study

AimSince March of 2020, tight measures against COVID-19, newly declared as pandemic by World Health Organization, affected almost all areas of our lives. Similarly, as with other aspects of community life, marked changes were expected to take place in the drug scene during and after the first lockdown period (16.3.2020–14.4.2020 in Finland). Wastewater-based epidemiology (WBE) is an approach established as an objective means for monitoring regional drug use. In Finland, we began WBE in 2012 with ten cities, and ended up to 27 cities (approximately 60% of Finnish population) since 2020. This study illustrates regional trends in stimulant drug use in Finland during the COVID-19 pandemic as a part of trend monitoring for up to 10 years.MethodComposite 24-h samples of untreated wastewater were collected twice a year during one-week sampling periods from the inlets of wastewater treatment plants. Every other year 1-2 sampling campaigns were performed in the four biggest cities only. In 2020, two extra sampling periods (total of four periods) were carried out to evaluate the impact of measures against COVID-19 on drug use. Samples were analyzed with a fully validated UHPLC-MS/MS system. Daily drug consumption was calculated from the measured concentrations of the drugs or their metabolites in wastewater.ResultsWhile the stimulant drug consumption has been steadily increasing in Finland during the years of WBE, in 2020 the overall trend leveled off, with drug use being in higher level than ever before. However, the changes observed after the first quarter of 2020 varied markedly in different cities, with deepest decrease in Mariehamn, Åland, and highest increases in Lappeenranta and Vihti. During 2021, consumption of amphetamine, the dominant stimulant drug in Finland, was reduced by half from March to November/December. Instead, cocaine use remained in its newly increased level in Helsinki metropolitan area, bringing consumption of these two drugs record close to each other.COVID-19 measures may have played a role in the leveling observed in overall stimulant drug use trend, but the changes varied regionally and remained minor and transient in most cities. It seems that drug use was less affected by lockdown in 2020, as well as other measures, than many other societal phenomena. The marked fall of amphetamine consumption observed after spring of 2021 is temporally related to large seizures of drugs, which may have exhausted drug supplies. However, it can be speculated whether COVID-19 measures may have, for their part, delayed the recovery of illicit market after the seizures. Although the overall national drug situation is by far dominated by the biggest cities with largest populations, this study with opposing drug use trends in different cities shows again the importance of including a range of cities with different sizes and geographical locations in monitoring programs, both for a realistic picture of the overall drug situation, and to be able to offer objective and comparable data to various actors of both national and local level.ConclusionDrug use was less affected by measures against COVID-19 than many other societal phenomena. To obtain a realistic picture of the multifaceted drug situation, both during normal and exceptional circumstances such as COVID-19, it is essential to include a representative and wide enough selection of cities in monitoring programs.

Wastewater surveillance for different classes of pharmaceutical drugs: Focus on psychotropic drugs and their metabolites

Wastewater screening analysis for the determination of pharmaceutical drugs belonging to different classes, with particular focus on psychotropic drugs and their metabolites. Wastewater based epidemiology (WBE) recently emerged as an essential and complementary methodology for the evaluation of pharmaceutical and illicit drugs prevalence, by means of quantification of either the parent drugs or their human-specific metabolites in wastewater. Furthermore, water pollution by pharmaceutical drugs is becoming an unavoidable environmental issue of emerging concern, making the water quality monitoring crucial to safeguard its healthy supply to citizens. The most commonly used pharmaceuticals include non-steroidal anti-inflammatory drugs, cardiovascular drugs, anti-depressant and antipsychotic drugs; their Active Pharmaceutical Ingredients (APIs) and some of their metabolites are known to partially survive the conventional process of wastewater treatment. Our study involves a wide screening analysis on wastewater samples entering and leaving various purification plants in the Piedmont region (Italy) to identify the prevalent pharmaceutical drugs and metabolites present in it and so to provide important information about the diffusion of compounds of toxicological interest. A total of 140 target compounds (pharmaceuticals of various therapeutic classes, benzodiazepines and some of their metabolites) were investigated in wastewater samples using only 30 mL aliquots instead of the typical 500 mL reported in the literature. The filtered samples (0.22 μm filters) were extracted and purified by SPE and subsequently 5 μL of the concentrated samples was analyzed with UHPLC-QTOF-HRMS and UHPLC-MS/MS protocols. Thanks to the combined use of the two instruments, a full scan on the exact mass and MS/MS fragmentation data were collected for parent drugs and their main metabolites. 61 out of 119 targeted pharmaceutical drugs and 6 out of 21 targeted metabolites of psychotropic pharmaceutical drugs were detected over a wide concentration interval, ranging from ng/L to μg/L. The highest concentration was 62 μg/L detected for paracetamol. The classes of antidepressants, benzodiazepines, antiepileptics, and antipsychotic required particular attention as they are known to be harmful to aquatic flora and fauna. Among these, citalopram (mean concentration 25.8 ng/mL), lorazepam (mean concentration 32.2 ng/mL), trazodone (mean concentration 17.0 ng/mL) and carbamazepine (mean concentration 134 ng/mL) were detected in both the purifier's affluent and effluent waters. Since some environmental studies have shown that carbamazepine is one of the most frequently detected drugs in wastewater treatment plants, the presence of its main metabolites was explored. In particular, the presence of 10,11-dihydro-10-hydroxycarbamazepine, 10,11-dihydro-10,11-dihydroxycarbamazepine, oxarbamazepine and 10,11-dihydro-10,11-epoxycarbamazepine was revealed. The latter compound (approximately three times more concentrated than carbamazepine) proved to possess antiepileptic properties similar to carbamazepine, possibly producing neurotoxic effects. A reliable wastewater method of analysis involving a low sample volume, its SPE purification, and UHPLC-QTOF-HRMS and UHPLC-MS/MS targeted detection was developed for the simultaneous determination of 119 pharmaceutical drugs and 21 metabolites. The collected data are consistent with those available in the literature and confirm that many pharmaceutical drugs are present in wastewater at a level that pose some risk to flora and fauna, taking into account also the health issues associated with long-term simultaneous exposure to a large number of pharmaceutical products. In conclusion, wastewater surveillance is essential not only to identify the pharmaceutical drugs used in the area but also to monitor the purity of waters. In the future, analyses will be carried out to study the variation in the substances (including NPS) found over time and in the different territories at both intra- and inter-regional levels.

Prevalence and emergence of steroidal and non-steroidal anabolic agents in the Australian community measured through wastewater analysis

Develop and validate a method for the detection of performance- and image-enhancing drugs (PIEDs) in wastewater. Investigate prevalence of steroidal and non-steroidal anabolic agents in around 45% of the Australian population during Census week in August 2021. Explore changes and patterns in use over a 13-year period (2009–2021) and emergence of non-steroidal anabolic agents, e.g., selective androgen receptor modulators (SARMs), within one Australian wastewater treatment plant (WWTP) catchment. Performance- and image-enhancing drugs include, inter alia, steroidal (anabolic-androgenic steroids) and non-steroidal anabolic agents [e.g., SARMs, peroxisome proliferator-activated receptor delta (PPAR-δ) agonists, growth hormone secretagogues]. Despite known serious negative health side effects, these substances are misused by athletes and non-athletes to boost physical performance as well as appearance. Due to their misuse, often at concentrations significantly higher than for medical purposes, easy access through the internet, and lack of quality controls, PIED use has become a public health problem. The extent of this issue is difficult to quantify and is currently monitored through the combination of multiple data sources such as surveys, seizures, and anti-doping. To fully understand the prevalence of these substances, especially among the general population, a broader monitoring tool is needed to complement current sources of data. In the past two decades, wastewater-based epidemiology has been developed to investigate exposure to, use, and prevalence of chemicals within entire populations and has potential to be a useful additional tool for PIED monitoring. A solid-phase extraction (SPE) liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and validated for the detection and quantification of around 50 anabolic agents and metabolites in wastewater. This method was applied to wastewater samples collected in August 2021 from 51 WWTPs across Australia, covering roughly 11.6 million people. This method was also used for temporal analysis of anabolic agents in wastewater from one catchment collected from 2009 to 2021. Limits of quantification (LOQs) were, with few exceptions, in the low ng/L with steroids and their metabolites generally having higher LOQs than the non-steroidal anabolic agents. Spatial wastewater analysis showed the use of multiple types of PIEDs within the Australian population in August 2021. Results of temporal samples indicated no clear trends for detected steroidal anabolic agents. However, they showed that the first non-steroidal anabolic agent was detected as early as 2011, with others following in later years. After their first detection, all non-steroidal anabolic agent concentrations increased almost linearly until 2021, often with a peak in concentration in 2018. LOQs are suitable for the detection of trace amounts of PIEDs in wastewater, especially for SARMs and other non-steroidal anabolic agents. As the method is semi-quantitative, concentrations only give an indication of anabolic agent levels, however, these estimates are likely conservative. The analysis of wastewater across 51 Australian WWTPs revealed widespread use of multiple types of steroidal and non-steroidal anabolic agents. The steady increase of concentrations of non-steroidal anabolic agents in the temporal wastewater samples demonstrates the growing popularity of these substances within the general community. A suitable SPE LC-MS/MS method was developed for the analysis of steroidal and non-steroidal anabolic agents in wastewater influent and was successfully applied to spatial and temporal wastewater samples. Many non-steroidal anabolic agents were detected in wastewater for the first time. Spatial analysis showed the prevalence of anabolic agents and temporal analysis revealed the first detections and subsequent emergence of non-steroidal anabolic agents within an Australian WWTP catchment. Results demonstrated that wastewater analysis is a useful complementary tool to further our understanding of PIED use within the general population and that wastewater data may be useful to public health and PIED researchers.

Qualitative fingerprinting of psychoactive pharmaceuticals, illicit drugs, and related human metabolites in wastewater: A year-long study from Riga, Latvia

The COVID-19 pandemic has become an unprecedented public health emergency causing immense societal and socio-economic consequences. Multiple studies have outlined that interventions to curb the spread of the virus are likely to have an effect on substance use patterns. In this study, we explored the presence of psychoactive pharmaceuticals, illicit drugs and related human metabolites in 24-h composite wastewater samples that were collected weekly in 2021 from the central WWTP of Riga, Latvia. The analysis was performed via suspect screening approach using three separate high-resolution mass spectrometry (HRMS) workflows, which relied on reversed-phase liquid chromatography (RPLC), hydrophilic interaction liquid chromatography (HILIC) and direct infusion HRMS. In total, 39 out of 149 substances were detected throughout the sampling period. These include pharmaceuticals (mainly antiepileptics, antidepressants and antipsychotics), illicit drugs (e.g., MDMA, MDEA, cocaine, etc.) and new psychoactive substances (alpha-PVP). The results were evaluated in relation to COVID-19 incidence rate and the severity of containment and closure policies. For some compounds we observed temporal changes that may be potentially linked to the state of the pandemic. For instance, higher detection rates were observed for several illicit drugs during periods, when restrictions on public events were relaxed. Meanwhile, some psychoactive pharmaceuticals and drugs used to treat upper respiratory tract infections displayed increased prevalence in weeks when the national COVID-19 incidence rates were higher. However, without baseline reference data from previous years, it is difficult to discern how much of the relationships seen are linked to pandemic progression and seasonal variability.

Rapid, Multianalyte Detection of Opioid Metabolites in Wastewater.

By monitoring opioid metabolites, wastewater-based epidemiology (WBE) could be an excellent tool for real-time information on the consumption of illicit drugs. A key limitation of WBE is the reliance on costly laboratory-based techniques that require substantial infrastructure and trained personnel, resulting in long turnaround times. Here, we present an aptamer-based graphene field effect transistor (AptG-FET) platform for simultaneous detection of three different opioid metabolites. This platform provides a reliable, rapid, and inexpensive method for quantitative analysis of opioid metabolites in wastewater. The platform delivers a limit of detection 2-3 orders of magnitude lower than previous reports, but in line with the concentration range (pg/mL to ng/mL) of these opioid metabolites present in real samples. To enable multianalyte detection, we developed a facile, reproducible, and high-yield fabrication process producing 20 G-FETs with integrated side gate platinum (Pt) electrodes on a single chip. Our devices achieved the selective multianalyte detection of three different metabolites: noroxycodone (NX), 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), and norfentanyl (NF) in wastewater diluted 20× in buffer.

Stability of Human Stress Hormones and Stress Hormone Metabolites in Wastewater Under Oxic and Anoxic Conditions

Stability of Human Stress Hormones and Stress Hormone Metabolites in Wastewater Under Oxic and Anoxic Conditions