Search Results (84)

Background: New psychoactive substances (NPS) represent an evolving component of global substance use patterns and may contribute to HIV transmission through both injection-related and sexual risk behaviors. In Kazakhstan, where HIV incidence has increasingly shifted toward sexual transmission, evidence on HIV testing among NPS users remains limited. This study examined behavioral, social, and structural factors associated with HIV testing in this population. Methods: A cross-sectional study was conducted among 1500 adults reporting NPS use across six regions of Kazakhstan. Data were collected using structured interviewer-administered questionnaires. The primary outcome was self-reported HIV testing (ever tested: yes/no). Independent variables included sociodemographic characteristics, substance use behaviors, sexual practices, peer communication about HIV, and structural access to prevention services. Univariable logistic regression with Bonferroni correction (p < 0.001) was used for variable screening. Multivariable logistic regression models estimated adjusted odds ratios (AORs) with 95% confidence intervals (CIs). Model discrimination was assessed using the area under the receiver operating characteristic curve (AUC). Results: Overall, 86.7% of participants reported prior lifetime HIV testing. In the multivariable model (n = 1482), older age was associated with higher odds of testing (AOR 1.06 per year; 95% CI 1.04–1.08; p < 0.001). Compared with participants holding a bachelor’s degree or higher, those without a high school diploma had lower odds of testing (AOR 0.50; 95% CI 0.28–0.89). Injectable psychostimulant use was also associated with testing (AOR 1.40; 95% CI 1.21–2.01). Participants who never discussed HIV within peer networks were less likely to have been tested (AOR 0.69; 95% CI 0.49–0.97). Engagement with HIV prevention services (AOR 0.54; 95% CI 0.39–0.75) and use of prevention centers (AOR 0.63; 95% CI 0.45–0.87) were significantly associated with testing. The model demonstrated acceptable discrimination (AUC = 0.725). Conclusions: Lifetime HIV testing uptake among NPS users in Kazakhstan is high but influenced by educational attainment, peer communication, injection practices, and engagement with prevention services. Strengthening integration of prevention services and expanding peer-based outreach may improve equitable access to HIV testing in this population. Full article

Background/Objectives: Synthetic stimulants represent the most prevalent subclass on the new psychoactive substances (NPSs) market. However, the toxicokinetic properties of M-ALPHA, a regioisomer of MDMA and N-methyl-cyclazodone a pemoline derivative, are not yet characterized. Methods: Therefore, this study investigated the metabolism of both NPSs in pooled liver S9 fraction and rat urine, characterized cytochrome P450 (CYP) kinetics and plasma protein binding (PPB), and assessed the CYP inhibition potential of M-ALPHA, using high-performance liquid chromatography coupled to high resolution tandem mass spectrometry (HPLC-HRMS/MS). Results: Four metabolites of M-ALPHA were detected including one phase I and three phase II metabolites, resulting from demethylenation followed by subsequent methylation or glucuronidation. For N-methyl-cyclazodone, one phase I metabolite formed via N-demethylation was identified. The primary enzymes involved in M-ALPHA metabolism were CYP2B6 and CYP2D6. Notably, M-ALPHA inhibited these enzymes to a strong or moderate extent, respectively. In contrast, the metabolism of N-methyl-cyclazodone was primarily mediated by CYP2A6. PPB studies indicated low-to-moderate binding for both compounds, suggesting that significant protein-binding interactions are unlikely. Conclusions: As M-ALPHA only formed metabolites that overlapped with those of MDMA, differing only by minor retention time shifts, reliable HPLC-HRMS/MS-based identification may be challenging in clinical and forensic toxicology settings as well as doping analysis. Furthermore, drug–drug interactions following polydrug use cannot be excluded for either NPS, particularly when co-ingested with other CYP substrates metabolized by the same isoforms. Full article

The continuous emergence of New Psychoactive Substances (NPS) poses a significant challenge to public health and forensic toxicology due to their unpredictable pharmacology and rapid turnover on the illicit market. This study describes the development and validation of a high-resolution screening method for the simultaneous detection of 90 NPS in oral fluid (OF), a matrix of choice for non-invasive sampling and roadside testing. The analytical workflow utilizes a “dilute-and-shoot” approach (1:2 v/v dilution) followed by ultra-high-performance liquid chromatography coupled with a quadrupole-Orbitrap hybrid mass spectrometer (UHPLC-HRMS/MS). Chromatographic separation was achieved in 11 min using a biphenyl column and a gradient elution. The method was validated according to ANSI/ASB Standard 036 guidelines, covering 90 substances including synthetic cannabinoids (e.g., HHC, MDMB-4en-PINACA), synthetic cathinones, and high-risk synthetic opioids such as nitazenes and fentanyl analogues. Results showed high sensitivity, with limits of identification (LOI) reaching 1 ng/mL for 44.4% of the analytes and 5 ng/mL for 37.8%, while the remaining compounds showed higher LOIs ranging from 10 to 100 ng/mL. No significant matrix interference or carryover was observed. The method was successfully applied to real samples from external quality control programs and forensic cases. This robust and versatile screening tool is suitable for clinical and forensic applications, supporting the monitoring of emerging NPS trends. Full article

The rapid emergence of New Psychoactive Substances (NPS), particularly pyrrolidinophenone derivatives, poses a significant challenge for public health and forensic toxicology. While their neuropharmacological profiles as dopamine transporter inhibitors are well-documented, their cardiac toxicity remains poorly understood. This study employs a multiparametric New Approach Methodology (NAM) using zebrafish embryos to integrate neurobehavioral and cardiotoxic endpoints for comparative hazard prioritization. We evaluated nine pyrrolidine-containing cathinones, including α-PVP, MDPV, α-PiHP, MDPiHP, α-D2PV, 3-Cl-, 4-Cl-, and 3,4-Cl-α-PVP, and 4-F-3-Me-α-PVP, on locomotor activity and cardiac rhythmicity using high-speed video microscopy and dynamic pixel analysis. Across the series, compounds induced concentration-dependent negative chronotropy and, in most cases, locomotor suppression. Crucially, we identified a functional dissociation between atrial rate control and atrioventricular (AV) conduction. The 3,4-dichloro substitution (3,4-Cl-α-PVP) was the most potent inducer of negative chronotropy (EC₅₀ = 52.6 μM), whereas 4-Cl-α-PVP exhibited a distinct pro-arrhythmic liability, increasing the incidence of 2:1 AV block. Time-course locomotor profiling indicated that α-PVP and chlorinated analogs were among the most potent behavioral modifiers. Using a Functional Safety Index (AV block EC₅₀/locomotor EC₅₀-like), we show that most compounds exhibit wide separations between neurobehavioral inhibition and severe conduction impairment, while specific substitutions, particularly para-chlorination, are associated with comparatively reduced functional separation between these endpoints within the assay. Overall, these data demonstrate that subtle structural changes within the pyrrolidinophenone scaffold can shape distinct arrhythmic phenotypes and functional safety profiles, supporting zebrafish-based integrated screening as a rapid platform for prioritizing emerging synthetic cathinones with comparatively higher cardiac liability within this experimental framework. Full article

Introduction: The availability of toxicokinetic data is critical for detecting and monitoring the intake of psychoactive substances. Timely characterization of novel psychoactive substances (NPS) is particularly important to assess their abuse potential and inform public health responses. Methods: Toxicometabolomics offers a powerful approach to characterize xenobiotic metabolism through high-resolution profiling of biochemical transformations. It thus allows the finding of exogenous biomarkers, such as new drug metabolites, and endogenous biomarkers, which could be indications of acute drug ingestions or sample manipulation, as well as offering information on the mode of action of drugs. In this study, we applied a liquid chromatography–high-resolution mass spectrometry workflow to investigate the toxicometabolomics of two N1-sulfonated N,N-dimethyltryptamine derivatives with potential for both therapeutic use and recreational abuse. Results: Zebrafish (Danio rerio), an increasingly valuable model for preclinical pharmacology and toxicology studies, along with pooled human liver S9 fractions were used to elucidate metabolic pathways and identify key phase I and phase II biotransformations. Furthermore, untargeted metabolomics revealed significant downregulation of L-threonine associated with compound exposure. Conclusions: These findings advance the current understanding of tryptamine metabolism and underscore the utility of toxicometabolomics in the analytical evaluation of NPS. Full article

In addition to well-known traditional synthetic illicit drugs like cocaine, amphetamines, and heroin, an increasing number of new psychoactive substances (NPS) are appearing on the global drug market. Among them, cathinones represent a prominent class. These amphetamine-like compounds contain a stereogenic center, resulting in the possible presence of two enantiomers. Pure enantiomers of cathinone derivatives are not commonly available, and their production is cost-intensive. Thus, there is very little knowledge about the possible distinct effects of single enantiomers of cathinones. The objective of this study was to evaluate the stability of a set of eight cathinone derivatives, namely 3-methylethcathinone, 3-methylmethcathinone, 4-methylethcathinone, 4-methylmethcathinone, ethylone, 3,4-trimethylene-α-ethylaminovalerophenone, 3,4-tetramethylene-α-pyrrolidinovalerophenone, and 3,4-trimethylene-α-pyrrolidinobutiophenone, over a six-month period. Any racemization that may have occurred under different storage and solution conditions was monitored and compared. Pure enantiomeric fractions were collected on a multi-milligram scale using semi-preparative HPLC under isocratic normal-phase conditions. A Phenomenex Lux^® i-Cellulose-5, 5 μm 250 × 10 mm column containing cellulose tris(3,5-dichlorophenylcarbamate) served as the chiral selector. The tests showed that aqueous conditions, pH, temperature, chemical structure, sunlight, and oxygen influence compound stability. The long-term storage of cathinone derivative enantiomers was found to be optimal as solids under deep-freezing conditions or in a slightly acidified solvent where they are protected from air and light. Full article

Synthetic cannabinoids (SCs) represent one of the rapidly growing groups of new psychoactive substances (NPS) on the illicit drug market. SCs mimic the effects of Δ⁹-tetrahydrocannabinol, but they have a greater affinity to the receptors, resulting in more potent psychoactive effects than traditional substances. The toxicity and high abuse potential of SCs could pose serious health risks to their users. The challenges posed by the SCs require innovative monitoring strategies like the analysis of untreated wastewater, known as wastewater-based epidemiology (WBE). In this review article, we summarized the available literature on the detection and quantification of SCs in raw wastewater samples published between 2013 and 2025. We paid special attention to challenges related to different experimental stages of WBE analysis that hinder the accurate measurement of SCs and their metabolites. The reviewed studies show that wastewater analysis reflected the dynamic evolution of the illicit SCs market. As studies on the analysis of SCs in wastewater remain scarce, large monitoring campaigns and research performed in more locations are needed. Modern analytical hyphenated systems such as LC-MS are essential for the sensitive and accurate quantification of SC biomarkers in wastewater and their sound identification. Future studies should address further stability tests, investigation of SC metabolism, and careful selection of the effective SC extraction method from the complex environmental matrix. Full article

The rapid emergence of novel psychoactive substances (NPSs) after 2020 has created one of the most dynamic analytical challenges in modern forensic science. Hundreds of new synthetic cannabinoids, synthetic cathinones, synthetic opioids, hallucinogens, and dissociatives, appearing as hybrid or structurally modified analogues of conventional drugs, have entered the illicit market, frequently found in complex polydrug mixtures. This review summarizes recent advances in gas chromatography-mass spectrometry (GC-MS) for their detection, structural elucidation, and differentiation between 2020 and 2025 based on the ScienceDirect and Google Scholar databases. Due to its reproducible electron-ionization spectra, established reference libraries, and robustness toward complex matrices, GC-MS remains the primary tool for the separation and identification of emerging NPS. The current literature highlights significant improvements in extraction and pre-concentration procedures, derivatization strategies for thermally unstable analogues, and chromatographic optimization that enable discrimination between positional and stereoisomers. This review covers a wide range of matrices, including powders, herbal materials, vaping liquids, and infused papers, as well as biological specimens such as blood, urine, and hair. Chemometric interpretation of GC-MS data now supports automated classification and prediction of fragmentation pathways, while coupling with complementary spectroscopic techniques strengthens compound confirmation. The review emphasizes how continuous innovation in GC-MS methodology has paralleled the rapid evolution of the NPS landscape, ensuring its enduring role as a reliable, adaptable, and cost-effective platform for monitoring emerging psychoactive substances in seized materials. Full article

In recent years, the expansion of the illicit market for Novel Psychoactive Substances (NPS) has resulted in the emergence of numerous synthetic recreational drugs specifically designed to evade legal control and analytical detection. Among these, nitazenes represent one of the most potent classes of new synthetic opioids, although information regarding their toxicological properties remains limited. The present study aimed to assess the genotoxic potential of four nitazenes: clonitazene, etonitazene, isotonitazene and metonitazene in human lymphoblastoid TK6 cells using a flow cytometric version of the In Vitro Mammalian Cell Micronucleus Test, following OECD Guideline No. 487. Cells were exposed to concentrations ranging from 12.5 to 100 μM, and cytotoxicity, cytostasis, and apoptosis were evaluated to identify appropriate doses for micronucleus frequency assessment. Vinblastine, a well-established mutagen, was included as positive control. Our findings demonstrated that clonitazene and isotonitazene exhibit mutagenic potential, suggesting an increased long-term risk of developing chronic degenerative diseases. Furthermore, the results revealed that structurally related molecules can induce markedly different cellular effects, underscoring the importance of compound-specific toxicological evaluations to achieve a comprehensive understanding of the risks associated with their illicit use—risks often presumed to involve only addiction or acute toxicity. Full article

The global rise in New Psychoactive Substances (NPS), particularly synthetic cathinones, poses significant public health concerns due to their association with toxicity and fatalities. With oral intake a common route of consumption, understanding their effects on the human intestinal epithelium is essential but remains poorly explored. This study investigates the impact of four synthetic cathinones—3-CIC (3-chloro-N-isopropylcathinone), 4-CIC (4-chloro-N-isopropylcathinone), 3-Cl-TBC (3-chloro-terc-butylcathinone, bupropion), and 4-Cl-TBC (4-chloro-terc-butylcathinone)—using Caco-2 cells, focusing on protein expression changes. The results revealed a reduced protein content, with 3-CIC producing the most significant effects, including the up-regulation of 40–50 kDa proteins. These findings suggest pathway disruptions requiring further mechanistic investigation. Full article

Narcotics trafficking is a fundamental part of organized crime, posing significant and evolving challenges for forensic investigations. Addressing these challenges requires rapid, precise, and scientifically validated analytical methods for reliable identification of illicit substances. Over the past five years, forensic drug testing has advanced considerably, improving detection of traditional drugs—such as tetrahydrocannabinol, cocaine, heroin, amphetamine-type stimulants, and lysergic acid diethylamide—as well as emerging new psychoactive substances (NPS), including synthetic cannabinoids (e.g., 5F-MDMB-PICA), cathinones (e.g., α-PVP), potent opioids (e.g., carfentanil), designer psychedelics (e.g., 25I-NBOMe), benzodiazepines (e.g., flualprazolam), and dissociatives (e.g., 3-HO-PCP). Current technologies include colorimetric assays, ambient ionization mass spectrometry, and chromatographic methods coupled with various detectors, all enhancing accuracy and precision. Vibrational spectroscopy techniques, like Raman and Fourier transform infrared spectroscopy, have become essential for non-destructive identification. Additionally, new sensors with disposable electrodes and miniaturized transducers allow ultrasensitive on-site detection of drugs and metabolites. Advanced chemometric algorithms extract maximum information from complex data, enabling faster and more reliable identifications. An important emerging trend is the adoption of green analytical methods—including direct analysis, solvent-free extraction, miniaturized instruments, and eco-friendly chromatographic processes—that reduce environmental impact without sacrificing performance. This review provides a comprehensive overview of innovations over the last five years in forensic drug analysis based on the ScienceDirect database and highlights technological trends shaping the future of forensic toxicology. Full article

In this study, two isomeric cathinone derivatives, N-butyl-norbutylone and N-ethylhexylone, seized on the illicit drug market in Poland, were described and characterized by various instrumental analytical methods. The compounds were characterized by electrospray ionization mass spectrometry, high-resolution mass spectrometry, gas chromatography–mass spectrometry, and nuclear magnetic resonance spectroscopy. The two investigated compounds were confirmed as 1-(2H-1,3-benzodioxol-5-yl)-2-(butylamino)butan-1-one and 1-(2H-1,3-benzodioxol-5-yl)-2-(ethylamino)hexane-1-one, both of which were cathinone derivatives available on the new psychoactive substances (NPS) market. The obtained analytical data should be useful for forensic and toxicological purposes for quick and reliable compound identification. Full article

Over the past decade, more than a thousand new psychoactive substances (NPSs) have emerged worldwide. This rapid proliferation of “designer drugs” poses significant challenges for drug control, forensic analysis, and public health. Artificial intelligence (AI) has increasingly been applied to address these challenges in NPS design and analysis. This review provides a comprehensive overview of AI methodologies—including deep learning, generative models, and quantitative structure–activity relationship (QSAR) modeling—and their applications in the synthesis, prediction, and identification of NPSs. We discuss how AI-driven generative models have been used to design novel psychoactive compounds and predict their pharmacological activity, how QSAR models can forecast potency and toxicological profiles, and how machine learning is enhancing analytical chemistry workflows for NPS identification. Special emphasis is placed on mass spectrometry (MS)-based techniques, where AI algorithms (e.g., for spectral prediction and pattern recognition) are revolutionizing the detection and characterization of unknown NPSs. A dedicated section examines the legal and regulatory implications of AI-generated psychoactive substances in the European Union (EU) and United States (USA), highlighting current policies, potential gaps, and the need for proactive regulatory responses. The review concludes with a discussion of the benefits and limitations of AI in this domain and outlines future directions for research at the intersection of AI, analytical chemistry, and drug policy. Full article

Introduction: Novel psychoactive substances (NPSs) are substances not controlled by the United Nations’ 1961 Narcotic Drugs and 1971 Psychotropic Substances convention, which pose a threat to public health. The use of NPSs is growing among recreational drug users. NPSs mimic the effects of the existing illegal drugs; they are used as substitutes for the traditional drugs of use. NPSs are commonly marketed as safe substances. NPS abuse is especially risky among vulnerable individuals, such as children and adolescents. The Aim: This study aims to analyze the knowledge and attitudes of primary and high school students regarding NPSs, determining the frequency and patterns of NPS use, and examine motivational factors for their consumption. Methodology: The questionnaire was employed to primary and secondary school students of the city of Novi Sad in November 2024. The data were analyzed using the methods of descriptive and inferential statistics in the statistical software package JASP 0.18.1.0. Results: A total of 1095 participants took part in the survey (53.6% males and 46.4% females). The age range of participants was 11–18 years (mean age 14.637 years). The majority of pupils lived in the city (70.5%). The most numerous students were students with the highest overall grade. The proportion of students who were familiar with NPSs was 38.3%, while 61.7% of them were not aware of their existence. Living in cities correlated positively with the NPS knowledge. The NPS risk awareness was notably low. The proportion of students who tried one or more novel drugs was 1.918%. Conclusions: The abuse of novel psychoactive substances is a growing concern, particularly among young individuals, requiring increased awareness and education on their risks. Educational systems should provide accurate information to prevent false beliefs, while policymakers must legally regulate new drugs. A coordinated approach is crucial for effective prevention, involving education, media, and support from different organizations. Future studies should focus on the impact of education on attitudes towards NPSs. Full article

Drug abuse presents a significant global health challenge as the illicit drug market progresses from classic drugs to a growing prevalence of New Psychoactive Substances (NPS), particularly synthetic cathinones, which, although illegal, are often falsely marketed as safe and legal alternatives. The rapid increase in the use of these drugs complicates the assessment of their safety and effects on human health. However, they pose unique toxicological concerns that remain largely uncharacterized. This study investigated the toxic effects of three synthetic cathinones, namely, methylone, pentedrone, and 4-methylethcathinone (4-MEC), using the model organism C. elegans. We assessed the impact of these substances on animal survival, development, reproductive behavior, and longevity. Our results showed that short-term exposure (24 h) to concentrations of 5.0 mM or higher significantly reduced animal survival rates, while prolonged exposure (72 h) led to more pronounced toxicity, significantly reducing survival rates at concentrations as low as 1.0 mM. Moreover, sublethal concentrations resulted in developmental arrest. Additionally, pentedrone impaired reproductive capacity, while 4-MEC significantly shortened C. elegans lifespan. These findings highlight the urgent need for further investigation into the implications of synthetic cathinone use on human health through in vivo models as their prevalence in the illicit drug market continues to rise. Full article