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The use of illicit drugs causes unquestionable societal and economic damage. To implement actions aimed at combating drug abuse, it is necessary to assess illicit drug consumption patterns. The purpose of this paper was to develop, optimize, validate and apply a procedure for determining new psychoactive substances (NPSs) and classic drugs of abuse and their main metabolites in wastewater samples by using solid phase extraction (SPE) and high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). Moreover, detailed validation of the procedure was conducted. The developed SPE–HPLC-MS/MS procedure (within the sewage-based epidemiology strategy) allowed for the simultaneous, selective, very sensitive, accurate (recoveries ≥ 80.1%) and precise (CV ≤ 8.1%) determination of new and classic psychoactive substances in wastewater samples. This study is characterized by new scientific elements, especially in terms of the freeze-thaw and post-preparative stability of the selected psychoactive substances. This is the first time that NPSs (mephedrone and ketamine), the main metabolites of heroin (6-acetylmorphine, 6-AM) and marijuana (11-nor-9-carboxy-Δ9-tetrahydrocannabinol, THC-COOH) have been detected and monitored in Poland. This study is also the first to corroborate the data available from the EMCDDA and EUROPOL report and indicates that the retail market for cocaine is expanding in Eastern Europe.

Objective We propose a novel approach for measuring tobacco use in a community through the chemical analysis of nicotine metabolites in urban wastewater. It offers frequent monitoring and ‘real-time’, ‘evidence-based’ estimates of tobacco consumption which may complement epidemiological surveillance systems normally repeated only every few years. Methods Two urinary metabolites of nicotine, namely cotinine and trans-3′-hydroxycotinine, were selected as biomarkers of tobacco consumption in urban wastewater. During smoking, a known amount of nicotine is absorbed and after metabolism excreted as metabolites in urine, ending up in the wastewater; quantitative analysis of the metabolites in the wastewater allows back-calculation of the nicotine collectively absorbed by the population producing the sewage and, indirectly, their tobacco use. Representative samples of wastewater were collected from sewage treatment plants in eight Italian cities and analysed by mass spectrometry. Mass loads of the metabolites were used to estimate nicotine consumption. Results Wastewater analysis in the cities under study was used to estimate the number of cigarettes smoked, in order to compare the results of this study with those obtained from population surveys. The number of cigarettes calculated with the two methods were closely comparable and wastewater analysis was sufficiently sensitive to confirm the differences in tobacco consumption between northern and southern Italy, previously described in population surveys. Conclusions The described approach can serve as a supplementary indicator of tobacco consumption in local communities. This approach can provide objective and updated information, which are useful to assess the efficacy of tobacco-control interventions, with the aim of designing and implementing effective tobacco control plans.

The present study investigates the applicability of the chemical analysis of wastewater to assess the use of doping substances by the general population and amateur athletes. To this end, an analytical methodology that can identify and quantify a list of 15 substances from the groups of anabolic steroids, weight loss products, and masking agents in wastewater has been developed. The method uses solid phase extraction to increase the detection sensitivity of the target analytes, expected to be present at very low concentrations (ng L−1 range), and decrease possible matrix interferences. Instrumental analysis is performed by liquid chromatography coupled to high-resolution mass spectrometry, allowing data acquisition in both full scan and tandem MS mode. The method has been successfully validated at two concentration levels (50 and 200 ng L−1) with limits of quantification ranging between 0.7 and 60 ng L−1, intra- and inter-day precision expressed as relative standard deviation below 15%, procedural recoveries between 60 and 160% and matrix effects ranging from 45 to 121%. The stability of the analytes in wastewater was evaluated at different storage temperatures illustrating the importance of freezing the samples immediately after collection. The application of the method to 24-h composite wastewater samples collected at the entrance of three wastewater treatment plants and one pumping station while different sport events were taking place revealed the presence in wastewater, and hence the use, of the weight loss substances ephedrine, norephedrine, methylhexanamine, and 2,4-dinitrophenol. The use of these stimulants was visible just prior and during the event days and in greater amounts than anabolic steroids or masking agents.Graphical abstractChemical analysis of untreated wastewater reveals the use of prohibited doping substances during amateur sport event

Domestic wastewater was treated by five constructed wetland beds in series. Dissolved organic matter (DOM) collected from influent and effluent samples from the constructed wetland was investigated using fluorescence spectroscopy combined with fluorescence regional integration (FRI), parallel factor (PARAFAC) analysis, and two-dimensional correlation spectroscopy (2D-COS). This study evaluates the capability of these methods in detecting the spectral characteristics of fluorescent DOM fractions and their changes in constructed wetlands. Fluorescence excitation–emission matrix (EEM) combined with FRI analysis showed that protein-like materials displayed a higher removal ratio compared to humic-like substances. The PARAFAC analysis of wastewater DOM indicated that six fluorescent components, i.e., two protein-like substances (C1 and C6), three humic-like substances (C2, C3 and C5), and one non-humic component (C4), could be identified. Tryptophan-like C1 was the dominant component in the influent DOM. The removal ratios of six fluorescent components (C1–C6) were 56.21, 32.05, 49.19, 39.90, 29.60, and 45.87 %, respectively, after the constructed wetland treatment. Furthermore, 2D-COS demonstrated that the sequencing of spectral changes for fluorescent DOM followed the order 298 nm → 403 nm → 283 nm (310–360 nm) in the constructed wetland, suggesting that the peak at 298 nm is associated with preferential tryptophan fluorescence removal. Variation of the fluorescence index (FI) and the ratio of fluorescence components indicated that the constructed wetland treatment resulted in the decrease of fluorescent organic pollutant with increasing the humification and chemical stability of the DOM.

Wastewater-based epidemiology (WBE) is a powerful tool to gather epidemiological insights at the community level, providing objective data on population exposure to harmful substances. A considerable portion of the human exposure to these potentially harmful chemicals occurs unintentionally, unlike substances such as pharmaceuticals, illicit drugs, or alcohol. In this context, this comprehensive review analyzes WBE studies focused on classes of organic chemicals to which humans are unintentionally exposed, namely organophosphorus flame retardants, per- and polyfluoroalkyl substances (PFAS), benzotriazoles and benzothiazoles, phthalates and terephthalates, benzophenones, pesticides, bisphenols, and parabens. The review highlights some advantages of WBE for public health surveillance, e.g., non-invasive analysis, predictive capability, nearly real-time data, population-wide insights, no ethical approval, and unbiased sampling. It also discusses challenges and future research directions in WBE regarding exposure to harmful chemicals from various sources. The review emphasizes the critical role of wastewater sampling, sample preparation, quality control, and instrumental analysis in achieving accurate and reliable results. Furthermore, it examines the selection of human biomarkers for WBE studies and explores strategies to link WBE with human biomonitoring (HBM), which together enhance both the precision and effectiveness of exposure assessments.

Wastewater-based epidemiology (WBE) is an ever-increasing discipline that enables researchers to track near-real-time data concerning the recreational use of illicit drugs. Community illicit drug use was estimated in eleven Turkish cities, using eighteen wastewater treatment plants, representing the metropolitan cities and rural areas with different socio-demographic characteristics. In this study, 24-h composite influent wastewater samples were collected for 1 week per season between March 2019 and December 2019. Heroin, amphetamine, methamphetamine, ecstasy, cocaine, and marijuana were covered as illicit drugs in this study. Solid-phase extraction, LC-MS/MS separation, identification, and quantification were used as the analytical methods. Overall results indicate that both cocaine and MDMA usage increased in all cities on weekends. More specifically, it was observed that the average amount of cocaine use regarding 11 cities was 14.7 mg/1000 person/day, while figures for other substances are as follows: 9.5 for amphetamine, 34.5 for methamphetamine, 38.4 for MDMA, 42.2 for heroin, and lastly 5412 mg/1000p/day for marijuana. This study holds the position of being the most comprehensive one conducted, considering spatial and temporal datasets on illicit drug consumption obtained via WBE in Turkey.

An automated microextraction by packed sorbent followed by liquid chromatography–tandem mass spectrometry (MEPS-LC–MS/MS) method was developed for the determination of four endocrine disruptors—parabens, benzophenones, and synthetic phenolic antioxidants—in wastewater samples. The method utilizes a lab-made repackable MEPS device and a multi-syringe robotic platform that provides flexibility to test small quantities (2 mg) of multiple extraction phases and enables high-throughput capabilities for efficient method development. The overall performance of the MEPS procedure, including the investigation of influencing variables and the optimization of operational parameters for the robotic platform, was comprehensively studied through univariate and multivariate experiments. Under optimized conditions, the target analytes were effectively extracted from a small sample volume of 1.5 mL, with competitive detectability and analytical confidence. The limits of detection ranged from 0.15 to 0.30 ng L −1 , and the intra-day and inter-day relative standard deviations were between 3 and 21%. The method’s applicability was successfully demonstrated by determining methylparaben, propylparaben, butylated hydroxyanisole, and oxybenzone in wastewater samples collected from the São Carlos (SP, Brazil) river. Overall, the developed method proved to be a fast, sensitive, reliable, and environmentally friendly analytical tool for water quality monitoring.

The estimation of medication use is based on the statistical data from pharmacies and hospitals. Excessive use or misuse of some compounds, especially psychoactive medications, has not yet been monitored in Slovakia. Wastewater analysis provides useful data about the medication use and misuse in individual regions. This study is focused on the analysis of 23 substances in the wastewaters of Slovakia. The monitoring programme has included stimulants, opioid and morphine derivatives, benzodiazepines, antidepressants, drug precursors and their metabolites. Urinary markers of these compounds were analysed at WWTP influent in seven regions (Bratislava, Košice, Zvolen, Banská Bystrica, Trenčín, Prešov a Piešťany) using LC-MS/MS technique. The analysis was performed from March to October 2013. The pattern in use of these compounds was also monitored. Tramadol and venlafaxine were found to be the most concentrated compounds among of all studied psychoactive pharmaceuticals. The highest specific loads of tramadol were detected in Piešťany (409 mg/day/1000 inhabitants) and Zvolen (366 mg/day/1000 inhabitants). There is a considerable number of spa facilities (hotels) situated in these cities and this fact contributes to a higher occurrence of these psychoactive compounds in respective wastewaters.

The proliferation of new psychoactive substances (NPS) has necessitated the development and improvement of current practices for the detection and identification of known NPS and newly emerging derivatives. High-resolution mass spectrometry (HRMS) is quickly becoming the industry standard for these analyses due to its ability to be operated in data-independent acquisition (DIA) modes, allowing for the collection of large amounts of data and enabling retrospective data interrogation as new information becomes available. The increasing popularity of HRMS has also prompted the exploration of new ways to screen for NPS, including broad-spectrum wastewater analysis to identify usage trends in the community and metabolomic-based approaches to examine the effects of drugs of abuse on endogenous compounds. In this paper, the novel applications of HRMS techniques to the analysis of NPS is reviewed. In particular, the development of innovative data analysis and interpretation approaches is discussed, including the application of machine learning and molecular networking to toxicological analyses.

Illicit drug use is a global problem imposing social, economic, and health burdens on society. Wastewater-based epidemiology is an approach based on calculating the consumption of substances in the target population by analyzing the concentrations of human metabolic excretion products of licit and illicit substances in wastewater. This study estimated the changes in illicit drugs (cocaine, amphetamine, methamphetamine, ecstasy (3,4-Methylenedioxymethamphetamine), heroin, and marijuana (THC)), alcohol, and nicotine consumption in Adana Province during Ramadan compared to normal periods using wastewater-based epidemiology. An overall decrease was observed during the Ramadan Period, most strongly for ecstasy (29%) followed by heroin (19%). For cocaine, the variation was the slightest (8.6%). The differences were statistically significant for ecstasy, heroin, nicotine, and alcohol but not for cocaine, amphetamine, methamphetamine, and marijuana. This study is the first to show changes in the use of illicit drugs, alcohol, and nicotine under the influence of religious beliefs. In addition, there is limited data about illicit drugs, alcohol and nicotine abusers/users in Ramadan. This study provides information on the literature on this subject. Graphical abstract