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.