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-IMSHRMS), 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-alpha-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

Analysis of polycyclic aromatic hydrocarbons (PAHs) and their derivatives in biochar treated stormwater

Polycyclic aromatic hydrocarbons (PAHs), oxygenated PAHs (oxy-PAHs) and nitrogen heterocyclic polycyclic aromatic compounds (N-PACs) are persistent and semi-volatile organic compounds primarily formed due to incomplete combustion of organic material or, in the case of the derivatives, through transformation reactions of PAHs. Their presence in the environment is ubiquitous and many of them have been proven carcinogenic, teratogenic, and mutagenic. These toxic pollutants can therefore pose a threat to both ecosystem and human health and urges for remediation strategies for PAHs and derivatives from water bodies. Biochar is a carbon-rich material resulting from the pyrolysis of biomass resulting in a very porous matter with high surface area for an enhanced interaction with chemicals. This makes biochar a promising alternative for filtering micropollutants from contaminated aquatic bodies. In this work, a previously developed and validated methodology for the analysis of PAHs, oxy-PAHs and N-PACs in surface water samples was adapted for its utilization in biochar treated stormwater with special emphasis on scaling down the solid-phase extraction as well as including an additional filtering step for the removal of particulate matter in the media. & BULL; Optimized extraction method for PAHs, oxy-PAHs and N-PACs from stormwater treated with biochar. & BULL; Biochar strongly impacts the stormwater matrix and, therefore, additional steps are required in the extraction methodology. & BULL; Solid-phase extraction combined with GC-MS have been used to analyse PAHs, oxy-PAHs and N-PACs in stormwater treated with biochar

Monitoring new psychoactive substances use through wastewater analysis: current situation, challenges and limitations

New psychoactive substances (NPSs) are compounds that produce similar effects to those induced by illicit drugs (IDs), such as cocaine, cannabis, and amphetamines, but are not strictly regulated by international conventions. The consumption of NPSs is a growing public health problem in many communities. However, there is little knowledge regarding the extent and actual use of these new substances. Monitoring NPS use is arduous, and therefore, different sources of information need to be used to get more insight of the prevalence and diffusion of NPS use. Analysis of pooled urine and wastewater (WW) shows strong potential, giving a different and complementary light on this issue, although presents some limitations and challenges that must be taken into account. Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) is one of the most powerful approaches for screening a large number of NPSs because of the accurate-mass full-spectrum acquisition measurements. By using a comprehensive and updated NPS database, LC-HRMS is flexible enough to confront the ever-changing NPS market. In this “current opinion”, we give our point of view on the usefulness of pooled urine and WW analysis and on the potential application of wastewater-based epidemiology as source of information for NPS use, explaining the main bottlenecks and future perspectives in this emerging research field

Development of a Retention Time Interpolation scale (RTi) for liquid chromatography coupled to mass spectrometry in both positive and negative ionization modes

The accuracy and sensitivity of high resolution mass spectrometry (HRMS) enables the identification of candidate compounds with the use of mass spectrometric databases among other tools. However, retention time (RT) data in identification workflows has been sparingly used since it could be strongly affected by matrix or chromatographic performance. Retention Time Interpolation scaling (RTi) strategies can provide a more robust and valuable information than RT, gaining more confidence in the identification of candidate compounds in comparison to an analytical standard. Up to our knowledge, no RTi has been developed for LC-HRMS systems providing information when acquiring in either positive or negative ionization modes. In this work, an RTi strategy was developed by means of the use of 16 isotopically labelled reference standards, which can be spiked into a real sample without resulting in possible false positives or negatives. For testing the RTi performance, a mixture of several reference standards, emulating suspect analytes, were used. RTi values for these compounds were calculated both in solvent and spiked in a real matrix to assess the effect of either chromatographic parameters or matrix in different scenarios. It has been demonstrated that the variation of injection volume, chromatographic gradient and initial percentage of organic solvent injected does not considerably affect RTi calculation. Column aging and solid support of the stationary phase of the column, however, showed strong effects on the elution of several test compounds. Yet, RTi permitted the correction of elution shifts of most compounds. Furthermore, RTi was tested in 47 different matrices from food, biological, animal feeding and environmental origin. The application of RTi in both positive and negative ionization modes showed in general satisfactory results for most matrices studied. The RTi developed can be used in future LC-HRMS screening analysis giving an additional parameter, which facilitates tedious processing tasks and gain more confidence in the identification of (non)-suspect analytes

Perspectives and challenges associated with the determination of new psychoactive substances in urine and wastewater – A tutorial

New psychoactive substances (NPS), often designed as (legal) substitutes to conventional illicit drugs, are constantly emerging in the drug market and being commercialized in different ways and forms. Their use continues to cause public health problems and is therefore of major concern in many countries. Monitoring NPS use, however, is arduous and different sources of information are required to get more insight of the prevalence and diffusion of NPS use. The determination of NPS in pooled urine and wastewater has shown great potential, adding a different and complementary light on this issue. However, it also presents analytical challenges and limitations that must be taken into account such as the complexity of the matrices, the high sensitivity and selectivity required in the analytical methods as a consequence of the low analyte concentrations as well as the rapid transience of NPS on the drug market creating a scenario with constantly moving analytical targets. Analytical investigation of NPS in pooled urine and wastewater is based on liquid chromatography hyphenated to mass spectrometry and can follow different strategies: target, suspect and non-target analysis. This work aims to discuss the advantages and disadvantages of the different data acquisition workflows and data exploration approaches in mass spectrometry, but also pays attention to new developments such as ion mobility and the use of in-silico prediction tools to improve the identification capabilities in high-complex samples. This tutorial gives an insight into this emerging topic of current concern, and describes the experience gathered within different collaborations and projects supported by key research articles and illustrative practical examples

Plan de intervención fisioterápico tras resección de artroplastia de cadera sin reimplantación: a propósito de un caso

Una de las patologías más discapacitantes que afectan a la población de edad avanzada es la fractura de cadera. El tratamiento de elección suele ser la artroplastia protésica, pero el 5-10% de ellas falla, normalmente por un proceso infeccioso. Ante esta situación, una opción de tratamiento es la resección de la artroplastia de cadera sin reimplantación, ante la cual, la fisioterapia constituye una herramienta fundamental para devolver a los pacientes su funcionalidad previa, en la medida de lo posible. El objetivo del trabajo es describir un plan de intervención fisioterápico, aplicado en un paciente intervenido de una resección de artroplastia de cadera sin reimplantación, para restaurar la funcionalidad del miembro inferior afecto y evaluar los resultados obtenidos. Dicha intervención se lleva a cabo durante un mes, y está centrada en la terapia manual, cinesiterapia, ejercicio terapéutico y rehabilitación de la marcha. Tras su aplicación se observa una reducción del dolor y una desaparición del edema de la rodilla. Además, se aumenta el rango de movimiento de cadera y rodilla y la fuerza muscular, lo cual conlleva una recuperación funcional, permitiendo la marcha con dos bastones y la subida y bajada de escaleras. En conclusión, tras la aplicación del plan de intervención se produjo una gran mejoría, especialmente en la funcionalidad del paciente, aunque sin llegar a alcanzar el estado previo a la cirugía. Habría sido necesario un mayor tiempo de tratamiento para seguir mejorando su funcionalidad e independencia debido a la naturaleza de la patología.<br /

Anatomical study about the variations in renal vasculature

Renal vascularization is classically described as a renal artery and vein. However, this vascular pattern presents numerous anatomical variations in terms of their number, origin and course due to ontogenetic alterations. The aim was to carry out a descriptive study of the renal vascular pattern observed during the dissection of cadavers intended for teaching purposes. A descriptive and observational study of renal vascular anatomy was carried out by dissecting 16 renal blocks from 8 cadavers donated to science and used for teaching at the Faculty of Medicine of the University of Zaragoza. The prevalence of arterial variations was 75% (56.3% for polar renal arteries, 12.5% for pre-hilar branching and 6.25% for double communicating arterial arch) and venous was 62.5% (12.5% for polar renal veins, 25% for late venous confluence, 6.25% for triple renal vein and 18.75% for double circumaortic renal vein). We conclude that the renal vascular anomalies occur with high frequency; for this reason, knowledge of these anomalies is extremely important for the correct planning of numerous medical-surgical activities

Prediction of Retention Time and Collision Cross Section (CCSH+, CCSH–, and CCSNa+) of Emerging Contaminants Using Multiple Adaptive Regression Splines

Ultra-high performance liquid chromatography coupled to ion mobility separation and high-resolution mass spectrometry instruments have proven very valuable for screening of emerging contaminants in the aquatic environment. However, when applying suspect or nontarget approaches (i.e., when no reference standards are available), there is no information on retention time (RT) and collision cross-section (CCS) values to facilitate identification. In silico prediction tools of RT and CCS can therefore be of great utility to decrease the number of candidates to investigate. In this work, Multiple Adaptive Regression Splines (MARS) were evaluated for the prediction of both RT and CCS. MARS prediction models were developed and validated using a database of 477 protonated molecules, 169 deprotonated molecules, and 249 sodium adducts. Multivariate and univariate models were evaluated showing a better fit for univariate models to the experimental data. The RT model (R2 = 0.855) showed a deviation between predicted and experimental data of ±2.32 min (95% confidence intervals). The deviation observed for CCS data of protonated molecules using the CCSH model (R2 = 0.966) was ±4.05% with 95% confidence intervals. The CCSH model was also tested for the prediction of deprotonated molecules, resulting in deviations below ±5.86% for the 95% of the cases. Finally, a third model was developed for sodium adducts (CCSNa, R2 = 0.954) with deviation below ±5.25% for 95% of the cases. The developed models have been incorporated in an open-access and user-friendly online platform which represents a great advantage for third-party research laboratories for predicting both RT and CCS data.Funding for open access charge: CRUE-Universitat Jaume

In vitro bioanalytical assessment of toxicity of wetland samples from Spanish Mediterranean coastline

Background Fresh water bodies represent less than 1% of overall amount of water on earth and ensuring their quality and sustainability is pivotal. Although several campaigns have been performed to monitor the occurrence of micropollutants by means of chemical analysis, this might not cover the whole set of chemicals present in the sample nor the potential toxic effects of mixtures of natural and anthropogenic chemicals. In this sense, by selecting relevant toxicity endpoints when performing in vitro bioanalysis, effect-based methodologies can be of help to perform a comprehensive assessment of water quality and reveal biological activities relevant to adverse health effects. However, no prior bioanalytical study was performed in wetland water samples from the Spanish Mediterranean coastline. Methods Eleven samples from relevant water bodies from the Spanish Mediterranean coastline were collected to monitor water quality on 8 toxicity endpoints. Aryl hydrocarbon receptor (AhR), androgenicity (AR+ and AR-), estrogenicity (ER+ and ER-), oxidative stress response (Nrf2) and vitamin D receptor (VDR+ and VDR-) reporter gene assays were evaluated. Results AhR was the reporter gene assay showing a more frequent response over the set of samples (activated by 9 out of 11 samples), with TCDD-eq in the range 7.7-22.2 pM. For AR, ER and VDR assays sporadic activations were observed. Moreover, no activity was observed on the Nrf2 reporter gene assay. Wastewater and street runaway streams from Valencia could be responsible for enhanced activities in one of the water inputs in the Natural Park 'L'Albufera'. Conclusions Water quality of relevant wetlands from the Spanish Mediterranean coastline has been evaluated. The utilization of a panel of 5 different bioassays to cover for different toxicity endpoints has demonstrated to be a good tool to assess water quality