Screening for new psychoactive substances in wastewater from educational institutions.

Drug (ab)use among young people is a serious issue, negatively impacting their well-being and prospects. The emergence of new psychoactive substances (NPS) further complicates the situation as they are easily accessible (e.g., online), but users are at high risk of intoxication as their chemical identity is often unknown and toxicity poorly understood. While surveys and drug testing are traditionally used in educational institutions to comprehend drug use trends and establish effective prevention programs, they are not without their limitations. Accordingly, we investigated the occurrence of NPS in educational institutions through wastewater analysis and critically evaluated the viability of the approach. The study included eight wastewater samples from primary schools (ages 6-15 years), six from secondary schools (ages 15-19 years), three from institutions for both secondary and higher education (ages 15+), and six from higher educational institutions (ages 19+). Samples were obtained mid-week and evaluated in two Slovenian municipalities; the capital Ljubljana and a smaller one (M1). Samples were screened using liquid chromatography-ion mobility-high-resolution mass spectrometry (LC-IMS-HRMS), and NPS identified at three levels of confidence (Level 1: unequivocal, Level 2: probable, Level 3: tentative) from a suspect list containing over 5600 entries. NPS were identified in all types of educational institutions. Most were synthetic stimulants, with 3-MMC, ephedrine, 4-chloro-α-PPP, and ethcathinone being unequivocally identified. Also, NPS were present in wastewater from all educational institution types revealing potential spatial but no inter-institutional trends. Although specific groups cannot be targeted, the study, as a proof-of-concept, demonstrates that a suspect screening of wastewater employing LC-IMS-HRMS can be used as a radar for NPS in educational institutions and potentially replace invasive drug testing.

Occurrence and sources of residues of drugs of abuse in an urban aquifer: Chemical analysis and solute transport modelling.

This study investigated the occurrence and potential sources of residues of drugs of abuse in an urban aquifer beneath the City of Ljubljana using water analysis and a solute transport model designed to predict nitrogen distribution. Samples were collected from three sources: 28 wastewater samples (24-h composites), 4 aquifer-recharging river samples (grab), and 22 groundwater samples. The samples were analysed for residues of commonly (ab)used licit drugs (nicotine and alcohol), medications of abuse (morphine, methadone, codeine, and ketamine), and illicit drugs (tetrahydrocannabinol - THC, cocaine, amphetamines, and heroin) using liquid-liquid (alcohol residue) and solid-phase extraction, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Additionally, we used solute transport modelling to predict the spatial distribution of drug residues in the aquifer and their potential sources. Nicotine (up to 45,7 ng/L), cotinine (up to 5.86 ng/L), trans-3'-hydroxycotinine (up to 0.528 ng/L) and benzoylecgonine (up to 0.572 ng/L) were the most commonly detected drug residues in groundwater, followed by cocaine (

Three years of wastewater surveillance for new psychoactive substances from 16 countries.

The proliferation of new psychoactive substances (NPS) over recent years has made their surveillance complex. The analysis of raw municipal influent wastewater can allow a broader insight into community consumption patterns of NPS. This study examines data from an international wastewater surveillance program that collected and analysed influent wastewater samples from up to 47 sites in 16 countries between 2019 and 2022. Influent wastewater samples were collected over the New Year period and analysed using validated liquid chromatography - mass spectrometry methods. Over the three years, a total of 18 NPS were found in at least one site. Synthetic cathinones were the most found class followed by phenethylamines and designer benzodiazepines. Furthermore, two ketamine analogues, one plant based NPS (mitragynine) and methiopropamine were also quantified across the three years. This work demonstrates that NPS are used across different continents and countries with the use of some more evident in particular regions. For example, mitragynine has highest mass loads in sites in the United States, while eutylone and 3-methylmethcathinone increased considerably in New Zealand and in several European countries, respectively. Moreover, 2F-deschloroketamine, an analogue of ketamine, has emerged more recently and could be quantified in several sites, including one in China, where it is considered as one of the drugs of most concern. Finally, some NPS were detected in specific regions during the initial sampling campaigns and spread to additional sites by the third campaign. Hence, wastewater surveillance can provide an insight into temporal and spatial trends of NPS use.

Removal of residues of psychoactive substances during wastewater treatment, their occurrence in receiving river waters and environmental risk assessment.

Continuous consumption combined with incomplete removal during wastewater treatment means residues of psychoactive substances (licit drugs, medications of abuse and illicit drugs) are constantly introduced into the aquatic environment, where they have the potential to affect non-target organisms. In this study, 17 drug residues of psychoactive substances were determined in wastewater influent, effluent and in receiving rivers of six Slovene municipal wastewater treatment plants employing different treatment technologies. Variations in removal efficiencies (REs) during spring, summer and winter were explored, and ecotoxic effects were evaluated using in silico (Ecological Structure-Activity Relationships software-ECOSAR) and in vivo (algal growth inhibition test) methods. Drug residues were detected in influent and effluent in the ng/L to µg/L range. In receiving rivers, biomarkers were in the ng/L range, and there was good agreement between measured and predicted concentrations. On average, REs were highest for nicotine, 11-nor-9-carboxy-∆9-tetrahydrocannabinol (THC-COOH), cocaine residues, and amphetamine (>90 %) and lowest for methadone residues (<30 %). REs were comparable between treatments involving activated sludge and membrane bioreactors, while the moving biofilm bed reactor (MBBR) removed cotinine, cocaine, and benzoylecgonine to a lesser extent. Accordingly, higher levels of nicotine and cocaine residues were detected in river water receiving MBBR discharge. Although there were seasonal variations in REs and levels of drug residues in receiving rivers, no general pattern could be observed. No significant inhibition of algal growth (Chlamydomonas reinhardtii) was observed for the tested compounds (1 mg/L) during 72 h and 240 h of exposure, although effects on aquatic plants were predicted in silico. In addition, environmental risk assessment revealed that levels of nicotine, methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), morphine, and 3,4-methylenedioxymethamphetamine (MDMA) pose a risk to aquatic organisms. Since nicotine and EDDP can have acute and chronic effects, the authors support regular monitoring of receiving surface waters, followed up by regulatory actions.

Enantiomeric profiling of amphetamines in wastewater using chiral derivatisation with gas chromatographic-tandem mass spectrometric detection.

Enantiomeric profiling can supplement wastewater-based epidemiology (WBE) by providing additional information on drug origin (licit or illicit), improving consumption estimates, i.e., differentiating between disposal and consumption, and offering an insight into the potency of drugs available on the illicit drug market. We report on the enantiomeric profiling of amphetamines in wastewater using R(-)-α-methoxy-α-(trifluoromethyl) phenylacetyl chloride (R-MTPCl), a chiral derivatising agent and GC-MS/MS. The method performed well when evaluated against the SANTE/12682/2019 guidelines in terms of recovery (81-99%), accuracy (99-111%), repeatability (1-8%RSD) and linearity (LOQ-1000 ng/mL). The LOD and LOQ were 120 ng/L and 400 ng/L, respectively. The method was applied to samples of raw wastewater from two Slovene municipalities with unusual levels of amphetamines: Ljubljana (LJ1) and Velenje (VE1). LJ1 had an anomalously high mass load of MDMA (3,4-methylenedioxymethamphetamine) identified during SCORE 2020, and VE1 is a representative sample of the consistently high mass load of amphetamine. A second Ljubljana sample (LJ2) was chosen as a representative sample. The presence of racemic MDMA (EF = 0.511) in LJ1 indicated the disposal of the unused drug into the sewer, while the enrichment of R-MDMA (EF = 0.666) in the combined extract sample from Ljubljana (LJ2) indicated consumption. In the case of Velenje and Ljubljana, it is impossible to distinguish between the direct disposal and consumption of amphetamine and methamphetamine. Also, since amphetamine/methamphetamine-based prescription medications are unavailable in Slovenia, racemic amphetamine in VE1 (EF = 0.514) and LJ2 (EF = 0.459) indicate racemic and the more potent S-amphetamine are sold on the illicit drug market. Only S-methamphetamine was detected in wastewater (LJ2: EF = 0), indicating the presence of only the more potent S-methamphetamine on the illicit drug market. Overall, enantiomeric profiling provided useful information on amphetamine residues. In addition, chiral derivatisation can be a cost-effective alternative to using chiral chromatographic columns for the enantiomeric profiling of amphetamines in wastewater.

Investigation of drugs of abuse in educational institutions using wastewater analysis.

Wastewater analysis was used to investigate drug prevalence in primary and secondary schools and institutes of higher education located in urban and non-urban areas of six municipalities in Slovenia. Seven-hour composite raw wastewater samples from 44 educational institutions, including 19 primary schools (6-15 yrs.), ten secondary schools (15-19 yrs.), nine higher education institutions (19+ yrs.) and six mixed secondary and higher education institutions (15+ yrs.), were collected at the end of the 2018/2019 academic year. Metabolic residues of licit drugs (nicotine and alcohol), medications of abuse (morphine, codeine and methadone) and illicit drugs (cannabis, cocaine, amphetamine, methamphetamine, ecstasy and heroin) were targeted in the study. The analysis was carried out using solid-phase extraction and direct injection combined ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Biomarkers of nicotine, alcohol and cannabis intake were the most frequently detected, indicating a high prevalence of these drugs. Morphine and codeine were also detected, while among the stimulants, benzoylecgonine had the highest detection frequency. Drug differences were found between different levels of educational institution, geographic location (inter-municipality comparison) and degree of urbanization. However, t-distributed stochastic neighbour embedding (t-SNE) revealed that the level of educational institution was the main factor influencing the differences in drug prevalence. Although a good agreement between data from this study and other studies implementing wastewater analysis was observed, there was a discrepancy with Slovenian epidemiological survey data. Finally, despite certain drawbacks of the method, its application to detect drug residues in educational institutions provides a non-invasive insight into drug use trends.

Data in brief: Dataset of residues of drugs of abuse in wastewaters from Educational Institutions.

Metabolic residue concentration data for two licit drugs (nicotine and alcohol), three medications of abuse (morphine, methadone and codeine) and six illicit drugs (cannabis, cocaine, amphetamine, methamphetamine, ecstasy and heroin) were obtained from raw wastewater samples collected from 44 Slovenian educational institutions are presented. Also, concentrations obtained at one secondary school during a preliminary study is provided. The wastewater samples were collected at the end of the 2018/2019 academic year using time proportional sampling and analysed for 16 drug residues, extracted using solid-phase extraction and analysed using ultra-performance liquid chromatography hyphenated to tandem mass spectrometry (UPLC-MS/MS). Residues of nicotine and alcohol were determined by direct injection of filtered wastewater onto the UPLC. Concentrations data were studied based on educational level (primary, secondary and tertiary) and institution type (secondary schools: gymnasiums, vocational and technical schools, multi-programme schools; higher education institutions: natural sciences and social sciences), geographic location (municipalities) and degree of urbanisation (urban and non-urban areas). Due to the large number of different educational institutions included in the study, provided datasets are valuable for further studies on drug consumption patterns among young people. Drug presence and prevalence data for primary schools (6-15 years) offer an objective insight into drugs present in the early stage of a young person's development and help establish effective prevention programs. More details on the study can be found in [1].