• Up to 47 sites in 16 countries monitored over three years. • Synthetic cathinones most commonly found class. • Highest loads for mitragynine, particularly in sites from the United States. • Some temporal and regional trends evident.

Illicit drug use presents a significant challenge to global health and public safety, requiring innovative and effective monitoring strategies. This study aimed to evaluate the current landscape of wastewater-based epidemiology (WBE) for monitoring illicit drugs in Europe, focusing on collaboration, current practices, and barriers, while identifying opportunities for improvement. Cross-sectional survey-based study. Coordinated by the Sewage Analysis CORe Group Europe (SCORE) and the European Union Drugs Agency (EUDA), two surveys were conducted in 2023 targeting researchers and stakeholders using WBE for illicit drugs. Data were analysed to identify trends, gaps, and opportunities for improving WBE implementation. The findings indicate a robust research infrastructure and diverse analytical methods among European institutions. Two-thirds of the participating countries reported using WBE data to inform policy. However, challenges persist, particularly in securing funding and coordination, as well as generating national estimates from multiple locations and addressing specific local policy needs. WBE has proven to be a valuable tool for monitoring illicit drug trends and informing drug policies. To unlock its full potential, sustained funding, methodological standardization, and enhanced cooperation are essential. This study provides critical insights into the European WBE landscape, offering a roadmap for strengthening the integration of actionable WBE data into public health and policy frameworks.

New psychoactive substances are a group of synthetic or naturally occurring drugs that mimic the effects of controlled illicit drugs. With limited information around potency, effects and health risks, there is international concern around their use and thus surveillance efforts are needed for public health. This study presents a global assessment of new psychoactive substances in influent wastewater from 52 sites across 20 countries during the 2022/2023 New Year period. Using solid-phase extraction followed by liquid chromatography-tandem mass spectrometry, 21 new psychoactive substances were detected with mitragynine, 3-methylmethcathinone, and eutylone being the most prevalent. Notably, 3,4-methylenedioxy-PV8 was identified for the first time in sites in Australia and the United States. A retrospective analysis of population normalised mass loads for 3-methylmethcathinone at European sites revealed downward trends in 2022/2023 sampling period compared to the previous years, suggesting a possible impact of regional scheduling measures. Additionally, this work demonstrates the stability of new psychoactive substances in loaded cartridges for up to four months when stored at -20 °C. These findings highlight the value of wastewater-based epidemiology for global monitoring of emerging new psychoactive substances threats and policy evaluation.

Origanum dubium, mainly grown in the Mediterranean region, is one of the less extensively studied species among the oregano class. Oregano species are recognized for their significant pharmaceutical properties, primarily attributed to carvacrol and other phenolic compounds. The goal of this study was to establish a sustainable method for the extraction of carvacrol, total phenolic, and total flavonoid compounds (TPC and TFC, respectively). Pulse-mode ultrasonic-assisted extraction (UPAE) was employed, using ethanol–water mixtures as green solvents, for the extraction of the bioactive compounds from the plant material. A Box–Behnken design (BBD) coupled with Response Surface Methodology (RSM) was applied to optimize the extraction process with respect to the extraction temperature, extraction time, ethanol-to-water ratio of the solvent and power amplitude of the ultrasonic processor. The responses of carvacrol (determined by HPLC-PDA), TPC, and TFC (determined by spectrometric methods) were evaluated by RSM. The statistical model identified the optimal extraction conditions, which were a combination of increased extraction temperature (70 °C) for 26 min with an intermediate ethanol–water ratio (60%) at the maximum processor’s power amplitude (100%). These conditions led to the optimal response of the three measured parameters. The optimized parameters represent a green and efficient approach to obtain bioactive-enriched extracts from Origanum dubium, suitable for potential applications in functional foods, preservatives, or other applications.

Numerous plants are rich in secondary metabolites, such as phenols, flavonoids and terpenoids, which are associated with various beneficial properties. Extraction methods reported in literature allow the extraction of such phytochemicals, however, robust analytical methods, capable for the simultaneous determination of multi-class metabolites, applicable to multiple plant matrices, remain essential. This work presents the development, validation and application of a simple, sensitive, multi-residue and versatile High-Performance Liquid Chromatography – Photo-Diode Array detector (HPLC-PDA) method for the simultaneous determination of eleven secondary plant metabolites, exhibiting a wide range of polarities, within a single analytical run. The proposed method demonstrated excellent linearity ( R ² > 0.99) in a wide range of concentrations for all analytes, high method sensitivity, with limits of quantification (LOQ) ranging from 0.02 to 1.2 μg mL ⁻¹ , high precision, with intra-day and inter-day precision values below 5 and 10% respectively and accuracy in the 90–110% for low and 95–105% range for intermediate and high concentrations. The analytes were found to remain stable, with changes within 5% of the initial response, over a 30-day refrigerated-storage. The method was successfully tested on four plant extracts (i.e. Origanum vulgare , Rosmarinus officinalis , Lavandula angustifolia and Camellia sinensis ) and analyte identity was further confirmed with mass spectrometry (MS). Between five and eight metabolites were detected and quantified in each analyzed extract, demonstrating the broad applicability of the method. Overall, the proposed method constitutes a reliable and cost-effective tool for simultaneous separation and determination of chemically diverse secondary plant metabolites, which can facilitate phytochemical profiling for complex plant matrices.