Number Of New Psychoactive Substances Research Articles

In vitro and in vivo metabolic study of three new psychoactive β-keto-arylcyclohexylamines.

Since the 2000s, an increasing number of new psychoactive substances have appeared on the illicit drug market. β-Keto-arylcyclohexylamine compounds play important pharmacological roles in anesthesia; however, because these new psychoactive substances have rapidly increasing illicit recreational use, the lack of detailed toxicity data are of particular concern. Therefore, analysis of their metabolites can help forensic personnel provide references and suggestions on whether a suspect has taken an illicit new psychoactive β-keto-arylcyclohexylamine. The present study investigated the in vitro and in vivo metabolism and metabolites of three β-keto-arylcyclohexylamines: deschloro-N-ethyl-ketamine, fluoro-N-ethyl-ketamine and bromoketamine. In vitro and in vivo models were established using zebrafish and human liver microsomes for analysis of Phase I and Phase II metabolites by liquid chromatography-high-resolution mass spectrometry. Altogether, 49 metabolites were identified. The results were applied for the subject urine samples of known fluoro-N-ethyl-ketamine consumer screen analysis in forensic cases. Hydroxy-deschloro-N-ethyl-ketamine, hydroxy-fluoro-N-ethyl-ketamine and hydroxy-bromoketamine were recommended as potential biomarkers for documenting intake in clinical and forensic cases.

DrogoFinder: An open-source graphical user interface for the identification of illicit drugs using infrared spectroscopy

The development of Free and Open-Source Software (FOSS) is being increasingly recognized as an indispensable resource for the development, validation, and replication of analytical methodologies and tools. In this context, the use of Attenuated Total Reflectance Infrared (ATR-IR) spectroscopy for the in-situ identification of drugs requires software for the treatment of spectroscopic data, including spectral quality controls, the subtraction of possible excipients, and the comparison with well-defined libraries that can be complex and limits its application on the field analysis by non-expert users. Here we describe a user-friendly FOSS for drug identification that can be operated in an intuitive way, including automatic features such as i) quality controls that warn the user from possible technical problems during spectrum acquisition (e.g. water vapour, contaminated background), ii) the subtraction of the contribution of excipients and other substances present on the sample, and iii) the creation of custom and updated cloud-based libraries to deal with the great number of new psychoactive substances (NPS) that are constantly being developed and introduced for their consume. The proposed application was tested by expert and non-expert users with spectra from 42 drug samples measured in situ in a music festival and 39 seized samples measured in a laboratory. Non-expert users were able to flag technical problems in the spectra and to perform excipient subtractions automatically, obtaining drug identifications that matched those ones obtained by reference methods and dedicated commercial software. We share both, the code, and a standalone Windows app to reach worldwide users and scientists interested in the identification of drugs.

Electron activated dissociation - a complementary fragmentation technique to collision-induced dissociation for metabolite identification of synthetic cathinone positional isomers

Over the last decade, a remarkable number of new psychoactive substances (NPS) have emerged onto the drug market, resulting in serious threats to both public health and society. Despite their abundance and potential toxicity, there is little information available on their metabolism, a crucial piece of information for clinical and forensic purposes. NPS metabolism can be studied using in vitro models, such as liver microsomes, cytosol, hepatocytes, etc. The tentative structural elucidation of metabolites of NPS formed using in vitro models is typically carried out using liquid chromatography combined with high-resolution tandem mass spectrometry (LC-HRMS2) with collision-induced dissociation (CID) as a fragmentation method. However, the thermally-excited ions produced with CID may not be sufficient for unambiguous identification of metabolites or their complete characterization. Electron-activated dissociation (EAD), a relatively new fragmentation approach that can be used to fragment singly-charged ions, may provide complementary structural information that can be used to further improve the confidence in metabolite identification. The aim of this study was to compare CID and EAD as fragmentation methods for the characterization and identification of synthetic cathinone positional isomers and their metabolites. The in vitro metabolism of 2-methylethcathinone (2-MEC), 3-methylethcathinone (3-MEC) and 4-methylethcathinone (4-MEC) was investigated with both CID and EAD methods using LC-HRMS2. Four, seven and six metabolites were tentatively identified for the metabolism of 2-MEC, 3-MEC and 4-MEC, respectively. Here, the metabolism of 3-MEC and 2-MEC is reported for the first time. The EAD product ion mass spectra showed different fragmentation patterns compared to CID, where unique and abundant product ions were observed in EAD but not in CID. More importantly, certain EAD exclusive product ions play a significant role in structural elucidation of some metabolites. These results highlight the important role that EAD fragmentation can play in metabolite identification workflows, by providing additional fragmentation data compared with CID and, thus, enhancing the confidence in structural elucidation of drug metabolites.

Cytotoxiciy test of cathinones in a human kidney cell model

The number of new psychoactive substances (NSP) is continuously growing, completely changing the recreational drug market. Of this wide variety of new substances, a great emphasis is placed on synthetic cathinones. These drugs are known as "bath salts" and appeared on the market as legal substitutes for illicit substances, such as 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) or cocaine. The use of these substances is often considered harmless, but has actually been linked to several cases of toxicity and deaths. Most toxicological studies to date involve the use of liver or brain in vitro models, with little toxicological information on renal damage. The kidney is one of the excretory organs of these drugs and is therefore exposed to toxicity, this work covers the preliminary study of nephrotoxicity caused by cathinones. For this, a model of human kidney cells (the immortalized cell line HK-2) was exposed for 24h to a wide range of concentrations (0.01-10 mM) of two synthetic cathinones (methylone and MDPV) and the cytotoxicity was evaluated by the assay of reduction of MTT.

First-Generation Synthetic Cathinones Produce Arrhythmia in Zebrafish Eleutheroembryos: A New Approach Methodology for New Psychoactive Substances Cardiotoxicity Evaluation.

The increasing number of new psychoactive substances (NPS) entering the illicit drug market, especially synthetic cathinones, as well as the risk of cardiovascular complications, is intensifying the need to quickly assess their cardiotoxic potential. The present study aims to evaluate the cardiovascular toxicity and lethality induced by first-generation synthetic cathinones (mephedrone, methylone, and MDPV) and more classical psychostimulants (cocaine and MDMA) in zebrafish embryos using a new approach methodology (NAM). Zebrafish embryos at 4 dpf were exposed to the test drugs for 24 h to identify drug lethality. Drug-induced effects on ventricular and atrial heart rate after 2 h exposure were evaluated, and video recordings were properly analyzed. All illicit drugs displayed similar 24 h LC50 values. Our results indicate that all drugs are able to induce bradycardia, arrhythmia, and atrial-ventricular block (AV block), signs of QT interval prolongation. However, only MDPV induced a different rhythmicity change depending on the chamber and was the most potent bradycardia and AV block-inducing drug compared to the other tested compounds. In summary, our results strongly suggest that the NAM presented in this study can be used for screening NPS for their cardiotoxic effect and especially for their ability to prolong the QT intervals.

Comparison of controlled drugs and new psychoactive substances (NPS) regulations in East and Southeast Asia

New psychoactive substances (NPS) are substances of abuse that easily evade existing controlled drug regulations. This study conducted a systematic review on controlled drug regulations and analyzed the numbers of new psychoactive substances (NPS) reported in six East and Southeast Asian countries in comparison to US and UK from 2009 to 2020. Generally, more NPS were reported in the US (551) and UK (400), compared to Japan (379), China (221), Singapore (142), South Korea (99), Malaysia (41), and Taiwan (35). Legislative mechanisms including the specific listing of individual substances, generic control of a family of substances, analogue control of similar substances, temporary bans of new substances were evaluated. In this review, countries that have adopted a combination of legislative mechanisms were able to identify higher numbers of NPS for regulatory control, such as the US, UK, Japan, Singapore, and South Korea. These findings can provide references to countries like Malaysia and Taiwan, to strengthen NPS-related regulations nationally. Countries in the East and Southeast Asian region should be encouraged to collaborate more closely and to implement additional legislative approaches most relevant to the regional NPS trends to bridge the regulatory gap and to prevent the spread of emerging NPS.

Modern Psychoactive Substances and Their Detection in Biomedical Samples

Scientific and technological advancement of the mid- and late twentieth century led to the creation of a large number of new psychoactive substances (NPS), diverse in composition and spectrum of action. The primary stages of fight the spread of NPS are awareness of their characteristics, as well as the possibility of detecting both the substances themselves and biomarkers of their use (metabolites) in human biological objects. The purpose of the work is to review psychoactive compounds and methods of their detection performed for diagnostic purposes. The paper presents brief characteristics of the most common NPS, as well as features of their metabolism in the human. It is shown that the analysis of biological specimens collected from susceptive drug users is difficult due to the low content of analytes, the matrix influences, metabolism and the formation of various artifacts. The general scheme of screening of biological objects assumes the presence of two stages – a preliminary express analysis performed using immunochemical test systems, and a confirmatory analysis performed by gas and liquid chromatography-mass spectrometry. The article discusses the possibilities and limitations of targeted and non-targeted screening. The subject of special consideration was the problems of using chromatography-mass spectrometry methods of NPS analysis in Russian – issues of accessibility of standard substances, search libraries, etc. The article also presents brief characteristics of individual families of NPS, such as stimulants, synthetic cannabinoids, synthetic opioids and hallucinogens. In addition, the article shows that in a number of foreign armies, in particular, in the Armed forces of Ukraine (AFU), drugs and psychostimulants are used to create «fearless soldiers». Thus, methadone, amphetamine and other psychoactive substances, as well as a psychotomimetic – a structural and pharmacological similarity of the prohibited BZ – were found in objects delivered from the positions of the AFU for analysis.

Arylcyclohexylamine Derivatives: Pharmacokinetic, Pharmacodynamic, Clinical and Forensic Aspects.

Since the 2000s, an increasing number of new psychoactive substances (NPS) have appeared on the drug market. Arylcyclohexylamine (ACH) compounds such as ketamine, phencyclidine and eticyclidine derivatives are of particular concern, given their rapidly increasing use and the absence of detailed toxicity data. First used mainly for their pharmacological properties in anesthesia, their recreational use is increasing. ACH derivatives have an antagonistic activity against the N-methyl-D-aspartate receptor, which leads to dissociative effects (dissociation of body and mind). Synthetic ketamine derivatives produced in Asia are now arriving in Europe, where most are not listed as narcotics and are, thus, legal. These structural derivatives have pharmacokinetic and pharmacodynamic properties that are sometimes very different from ketamine. Here, we describe the pharmacology, epidemiology, chemistry and metabolism of ACH derivatives, and we review the case reports on intoxication.

Synthesis of benzo-condensed cyclic cathinone derivatives

Recently, the number of new psychoactive substances (NPS), which are sold as legal alternatives to conventional or misused drugs, has been growing on the market. Via small chemical modifications in the basic structure of the already banned substances, new derivatives are made. Although they pose a real threat to human health, they do circumvent regulation system. This fact led us to formulate the aim of this work, which is the synthesis of cyclic derivatives of cathinones, substances that are derived from natural substance S-cathinone. All commercial reagents and solvents were used without additional purification. 1H NMR and 13 C NMR spectra measured for structure conformation were acquired on an Agilent 400-MR DDR2 spectrometer with CDCl3 or CH3OD as the solvent. Cyclic derivatives of cathinone with different size of benzo-condensed ring were prepared using different synthetic approaches optimised to maximum possible yields. The first intermediate with a six membered condensed ring was synthesised using a nucleophilic addition of an aryllithium on cyclobutanone to form corresponding cyclobutanol derivative. This alcohol was transferred to an α-tetralone system using a radical cyclization reaction with potassium peroxodisulfate and silver nitrate. A cycliclization reaction of differently substituted 4-arylbutanoic acids employing polyphosphoric acid was used as a second approach. Also, five and four membered condensed ring derivatives were synthesised using two different approaches. The first approach was based on Friedel-Crafts acylation, where phenylpropanoic and phenylethanoic acid were converted to chlorides using oxalyl chloride, then using aluminium (III) chloride as a catalyst the intramolecular ring-closing reaction was completed. The second approach was cyclization of acids by means of thionyl chloride and trifluoromethanesulfonic acid. The benzocycloheptane ring was prepared using all cyclization reactions mentioned above. All the above-described intermediates were subsequently subjected to the sequence of reactions to prepare the final derivatives. A reaction of cyclic ketones with isoamyl nitrite provided oximes which were reduced with zinc dust in acetic acid to form acetamides. These acetamides were hydrolysed using hydrochloric acid yielding corresponding amines. These amines were transformed to trifluoroacetamides, which were methylated using iodomethane and the final step, acid hydrolysis gave rise to the target N-methylated cyclic derivatives. Different approaches towards the synthesis of benzocondensed bicyclic ketones with various sizes of the aliphatic ring are presented. These compounds have been used as intermediates to prepare new cyclic cathinones and their formal metabolites (N-demethylated derivatives). All the novel substances are fully characterized and, moreover, the possibility of their chiral separation using HPLC is being studied to assign their absolute configuration and study the biological activity of enantiomers.

Photoluminescent carbon nanomaterials for sensing of illicit drugs: focus.

The growth of illicit drugs is a serious social problem, putting great pressure on law enforcement officers to screen numerous suspicious samples at crime scenes. Although commercial colorimetric kits are available, they are limited to common illicit drugs. With increasing numbers of new psychoactive substances in the market, accurate and rapid screening assays are highly demanded. Carbon dots (C-dots) are photoluminescent (PL) carbon nanomaterials with unique properties of excellent stability against salt and photo-irradiation, low blinking, and biocompatibility. They can be prepared easily through various routes from numerous precursors. This Focus provides examples of C-dots based PL assays for screening illicit drugs. The drugs induce PL changes of C-dots mostly through electron transfer and energy transfer. Liquid- and solid-phase PL assays of C-dots can be applied for in-field screening, with advantages of rapidity, low cost, selectivity, and minimum color interference, showing their great commercial potential.

Blood concentrations of new synthetic opioids.

Over the last decade, there has been a significant growth in the market and number of new psychoactive substances (NPS). One of the NPS groups that has grown rapidly in recent years, bringing a new set of problems, consists of new synthetic opioids. The extreme potency of these compounds poses a high risk of acute poisoning, as an overdose can cause respiratory depression. Most of the information regarding human pharmacokinetics of new opioids is based on toxicological case reports and the data on concentrations of new opioids in human blood are scarce. The interpretation of results usually requires a comparison to previously published cases; therefore, a referenced compilation of previously published concentration data would be useful. The data were collected by searching the PubMed and Scopus databases and by using the Google search engine. All the available data from articles and reports that measured new opioid concentrations in plasma, serum, or whole blood were included in the data analysis. The presented tables list the observed concentrations in fatal and nonfatal cases involving 37 novel synthetic opioids. Blood levels of new opioids are extremely difficult to interpret. Low blood concentrations of these substances do not rule out acute poisoning as their high potency creates a risk of respiratory depression even at low doses. Opioid tolerance, frequent presence of other drugs, and additional diseases make it impossible to define concentration ranges, especially the minimum fatal concentrations. This report provides quick access to the source articles quantifying novel synthetic opioids.

New Psychoactive Substances and Suicidality: A Systematic Review of the Current Literature.

Background and Objectives: Over the past twenty years a large number of new psychoactive substances (NPS) have entered and modified the recreational drug scene. Their intake has been associated with health-related risks, especially so for vulnerable populations such as people with severe mental illness, who might be at higher risk of suicidality or self-injurious behavior. This paper aims at providing an overview of NPS abuse and the effects on mental health and suicidality issues, by performing a literature review of the current related knowledge, thereby identifying those substances that, more than others, are linked to suicidal behaviors. Materials and Methods: A comprehensive and updated overview of the literature regarding suicidality and NPS categories has been undertaken. An electronic search was performed, including all papers published up to March 2021, using the following keywords “NPS” OR “new psychoactive substances” OR “novel psychoactive substances” OR “synthetic cannabinoids” OR “phenethylamines” OR “synthetic cathinones” OR “tryptamines” OR “piperazines” OR “new synthetic opioids” OR “designer benzodiazepines” AND (“suicide” OR “suicidality”) NOT review NOT animal on the PubMed, Cochrane Library, and Web of Science online databases. Results: Suicidality and self-injurious behavior appear to be frequently associated with some NPS such as cathinones, synthetic cannabinoids, and new synthetic opioids. The results are organized according to the substances recorded. Conclusion: The growing use of NPS has become a significant clinical issue, causing increasing concern and challenges for clinicians working in both mental health and emergency departments. Thus, considering the associations between NPS and suicidality or self-injurious behaviors, areas where suicide-prevention efforts and strategies might be focused are the early detection, monitoring, and restriction of NPS.

Zebrafish early life stages as alternative model to study ‘designer drugs’: Concordance with mammals in response to opioids

The number of new psychoactive substances (NPS) on the illicit drug market increases fast, posing a need to urgently understand their toxicity and behavioural effects. However, with currently available rodent models, NPS assessment is limited to a few substances per year. Therefore, zebrafish (Danio rerio) embryos and larvae have been suggested as an alternative model that would require less time and resources to perform an initial assessment and could help to prioritize substances for subsequent evaluation in rodents. To validate this model, more information on the concordance of zebrafish larvae and mammalian responses to specific classes of NPS is needed. Here, we studied toxicity and behavioural effects of opioids in zebrafish early life stages. Synthetic opioids are a class of NPS that are often used in pain medication but also frequently abused, having caused multiple intoxications and fatalities recently. Our data shows that fentanyl derivatives were the most toxic among the tested opioids, with toxicity in the zebrafish embryo toxicity test decreasing in the following order: butyrfentanyl>3-methylfentanyl>fentanyl>tramadol>O-desmethyltramadol>morphine. Similar to rodents, tramadol as well as fentanyl and its derivatives led to hypoactive behaviour in zebrafish larvae, with 3-methylfentanyl being the most potent. Physico-chemical properties-based predictions of chemicals' uptake into zebrafish embryos and larvae correlated well with the effects observed. Further, the biotransformation pattern of butyrfentanyl in zebrafish larvae was reminiscent of that in humans. Comparison of toxicity and behavioural responses to opioids in zebrafish and rodents supports zebrafish as a suitable alternative model for rapidly testing synthetic opioids.

Prevalence of Stimulant, Hallucinogen, and Dissociative Substances Detected in Biological Samples of NPS-Intoxicated Patients in Italy

ABSTRACT A number of new psychoactive substances (NPS) have been released in the last decade, and the list of NPS continues to grow. This paper reports a retrospective evaluation of the toxicological analyses in 1,445 suspected intoxication cases by psychostimulant, hallucinogen, and dissociative NPS occurring in hospitals across Italy from 2011 to 2019. The objectives of the study were to contribute to the monitoring of the NPS diffusion based on analytically confirmed intoxications, and to evaluate the importance of the clinical toxicological laboratory in the diagnosis of NPS intoxication. For at least one NPS of the considered classes, 246 patients (17.0%) tested positive. Forty-four different NPS were detected and a consistent turnover was observed during the nine-year period, especially regarding cathinones. Among the positive cases, 47.2% tested positive for dissociative NPS, with particular regard to ketamine. Hallucinogens (30.9%) was the second most frequent NPS involved. Stimulants were found in 20% of the positive cases with a considerable presence of cathinones. Findings confirm the dynamism of the NPS phenomenon, underline the importance of awareness of this new public health threat among health care professionals, and highlight the need for analytical confirmation for the identification of the drugs in forensic contexts.

High lifetime, but low current, prevalence of new psychotropic substances (NPS) use in German drug detoxification treatment young inpatients

BackgroundOver the last 15 years, a large number of new psychoactive substances (NPS) has been identified, with their use being associated with a range of acute medical and psychopathological complications. Conversely, NPS addictive liability levels have not been systematically assessed in clinical populations. Aims of the studyInvestigating the lifetime and current prevalence of NPS use in a sample of substance use disorder (SUD) patients admitted to an inpatient detoxification treatment centre. MethodsAssessment of previous/current NPS intake carried out with the means of standardised questionnaire based on the European version of Addiction Severity Index. ResultsSome 206 patients (males 77.1%; average age: 30.7 years-old; most typical diagnosis: opioid/polydrug dependence) participated to the survey. Roughly half (e.g. 111/206; 53.9%) of them reported a lifetime use of NPS, most typically synthetic cannabinoids. Conversely, the current prevalence of NPS use was 2.9%; no NPS dependence condition was diagnosed. Among NPS users, 56.3% reported severe side-effects such as heavy anxiety or psychotic experience, and 64% reported an aversion of ever using the respective NPS again, whilst 84.3% of those reporting a single NPS intake reported an aversion. DiscussionThe sharp contrast between lifetime prevalence of NPS use and prevalence of current use might be explained by the high frequency of severe side effects reported by NPS users.

Comparative Untargeted Metabolomics Analysis of the Psychostimulants 3,4-Methylenedioxy-Methamphetamine (MDMA), Amphetamine, and the Novel Psychoactive Substance Mephedrone after Controlled Drug Administration to Humans.

Psychoactive stimulants are a popular drug class which are used recreationally. Over the last decade, large numbers of new psychoactive substances (NPS) have entered the drug market and these pose a worldwide problem to human health. Metabolomics approaches are useful tools for simultaneous detection of endogenous metabolites affected by drug use. They allow identification of pathways or characteristic metabolites, which might support the understanding of pharmacological actions or act as indirect biomarkers of consumption behavior or analytical detectability. Herein, we performed a comparative metabolic profiling of three psychoactive stimulant drugs 3,4-methylenedioxymethamphetamine (MDMA), amphetamine and the NPS mephedrone by liquid chromatography-high resolution mass spectrometry (LC-HRMS) in order to identify common pathways or compounds. Plasma samples were obtained from controlled administration studies to humans. Various metabolites were identified as increased or decreased based on drug intake, mainly belonging to energy metabolism, steroid biosynthesis and amino acids. Linoleic acid and pregnenolone-sulfate changed similarly in response to intake of all drugs. Overall, mephedrone produced a profile more similar to that of amphetamine than MDMA in terms of affected energy metabolism. These data can provide the basis for further in-depth targeted metabolome studies on pharmacological actions and search for biomarkers of drug use.

Intoxication cases associated with the novel designer drug 3',4'-methylenedioxy-α-pyrrolidinohexanophenone and studies on its human metabolism using high-resolution mass spectrometry.

Among the increasing number of new psychoactive substances, 3',4'-methylenedioxy-α-pyrrolidinohexanophenone (MDPHP) belongs to the group of synthetic cathinones, which are the derivatives of the naturally occurring compound cathinone, the main psychoactive ingredient in the khat plant. Currently, only limited data are available for MDPHP, and no information is available on its human metabolism. We describe the toxicological investigation of nine cases associated with the use of MDPHP during the period February-June 2019. Serum MDPHP concentrations showed a high variability ranging from 3.3 to 140 ng/mL (mean 30.3 ng/mL and median 16 ng/mL). Intoxication symptoms of the described cases could not be explained by the abuse of MDPHP alone because in all cases the co-consumption of other psychotropic drugs with frequent occurrence of opiates and benzodiazepines could be verified. Therefore, the patients showed different clinical symptoms, including aggressive behaviour, delayed physical response, loss of consciousness and coma. Liquid chromatography-high-resolution mass spectrometry was successfully used to investigate the human in vivo metabolism of MDPHP using authentic human urine samples. The metabolism data for MDPHP were further substantiated by the analysis of human urine using gas chromatography-mass spectrometry (GC-MS, a widely used systematic toxicological analysis method appropriate for the toxicological detection of MDPHP intake), which revealed the presence of seven phase I metabolites and three phase II metabolites as glucuronides. GC-MS spectral data for MDPHP and metabolites are provided. The identified metabolite pattern corroborates the principal metabolic pathways of α-pyrrolidinophenones in humans.

Flexible high resolution-mass spectrometry approach for screening new psychoactive substances in urban wastewater

The number of new psychoactive substances (NPS) on the recreational drug market has increased rapidly in the last years, creating serious challenges for public health agencies and law enforcement authorities. Epidemiological surveys and forensic analyses to monitor the consumption of these substances face some limitations for investigating their use on a large scale in a shifting market. The aim of this work was to develop a comprehensive and flexible screening approach for assessing the presence of NPS in urban wastewater by liquid chromatography-high resolution mass spectrometry (LC-HRMS). Almost 200 substances were selected as “priority NPS” among those most frequently and recently reported by the Early Warning Systems (EWS) of different agencies and were included in the screening. Wastewater samples were collected from several cities all over Europe in 2016 and 2017, extracted using different solid-phase cartridges and analysed by LC-HRMS. The screening workflow comprised three successive analytical steps and compounds were identified and confirmed following specific criteria from the current guidelines. Thirteen NPS were identified at different confidence levels by using analytical standards or information from libraries and literature, and about half of them were phenethylamines. As far as we know, this is the first time that four of them (i.e. 3,4-dimethoxy-α-pyrrolidinovalerophenone, para-methoxyamphetamine, 2-phenethylamine and α–methyltryptamine) have been found in urban wastewater. The proposed screening approach was successfully applied in the largest NPS European wastewater monitoring, providing an innovative and easily adapted procedure for investigating NPS. In the light of current challenges and specific future research issues, this approach may complement epidemiological information and help in establishing measures for public health protection.

HighResNPS.com: An Online Crowd-Sourced HR-MS Database for Suspect and Non-targeted Screening of New Psychoactive Substances.

The number of new psychoactive substances (NPS) is constantly increasing. However, although the number might be large, most NPS have a low prevalence of use, so keeping screening libraries updated with the relevant analytical targets becomes a challenge. One way to ensure sufficient screening coverage is to use shared high resolution-mass spectrometry (HR-MS) databases, such as HighResNPS.com: a free, online, spreadsheet-format, crowd-sourced HR-MS database for NPS screening. The aims of this study were (i) to present the database to the scientific community and (ii) to verify that the HighResNPS database can be utilized in suspect screening workflows for LC-HR-MS instruments and software from four different instrument vendors. A sample was spiked with 10 NPS, and participating laboratories then analyzed the sample with their respective HR-MS vendor platforms and the HighResNPS database. The HighResNPS data were obtained via a spreadsheet converted to fit the import specifications of the different vendor platforms. Suspect screening was performed using LC-HR-MS vendor platforms from Thermo Fisher, Waters, Bruker and Agilent. All 10 NPS were identified in at least three workflows used for the four different vendor platforms. Multiple users have submitted data to HighResNPS for the same NPS, which resulted in multiple true-positive identifications for these NPS. Suspect screening with LC-HR-MS can be based on diagnostic fragment ions reported by users of different vendor platforms and can support NPS identification in biological samples and/or seizure analyses when no reference standard is available in-house. The present work clearly demonstrates that HighResNPS data is compatible with instruments and screening software from at least four different vendor platforms. The database can thus serve as a useful add-on in LC-HR-MS screening workflows.

In Vitro Metabolism and Hepatic Intrinsic Clearance of the Synthetic Cannabinoid Receptor Agonist JWH-122 and Its Four ω-Halogenated Analogues.

The number of new psychoactive substances (NPS) emerging on the illicit drug market has increased over the last decade. Halogenation of existing illicit drugs is a particular trend, with the purpose of both circumventing the law and altering the toxicodynamic and toxicokinetic profiles of the compounds. This study investigates the in vitro impact of JWH-122 ω-halogenation (fluoro, chloro, bromo and iodo) on the metabolism, apparent intrinsic hepatic clearance and analytical targets for detecting drug consumption. Metabolite profiling was conducted with pooled human liver microsomes, suspended rat hepatocytes and pooled human hepatocytes. The in vitro half-life was also determined in pooled human hepatocytes. All samples were analysed by liquid chromatography/high-resolution mass spectrometry. All compounds, except for JWH-122, showed high formation rates of phase I metabolites, predominantly ω-COOH and methylnaphthyl hydroxylation metabolites. Phase II metabolites were ω-O-glucuronides, methylnaphthyl O-glucuronides and ω-glutathione conjugates. The relative ion intensity of the glutathione conjugates increased with the ω-halogen size, with I-JWH-122 having the highest intensity. Stability studies gave a low half-life and a high intrinsic hepatic clearance for JWH-122 (1305mL/min/kg) and MAM-2201 (1408mL/min/kg). Cl-, Br- and I-JWH-122 showed increasing half-life with increasing ω-halogen size, with intrinsic clearance values of 235-502mL/min/kg. The recommended analytical targets for consumption of JWH-122 or ω-halogenated JWH-122 analogues are the ω-COOH metabolites for unspecific profiling and the methylnaphthyl hydroxylated metabolites to distinguish the compounds. Furthermore, ω-halogenation with larger halogens appears to increase the intrinsic hepatic stability, thereby prolonging exposure and possibly the duration of action.