Number Of Synthetic Cannabinoids Research Articles

Simultaneous determination of five indole/indazole amide-based synthetic cannabinoids in electronic cigarette oil by ultra performance liquid chromatography

Synthetic cannabinoids (SCs), which are considered some of the most widely abused new psychoactive substances available today, are much more potent than natural cannabis and display greater efficacy. New SCs can be developed by adding substituents such as halogen, alkyl, or alkoxy groups to one of the aromatic ring systems, or by changing the length of the alkyl chain. Following the emergence of the so-called first-generation SCs, further developments have led to eighth-generation indole/indazole amide-based SCs. Given that all SCs were listed as controlled substances on July 1, 2021, the technologies used to detect these substances must be quickly improved. Due to the sheer number of SCs, the chemical diversity and the fast update speed, it is challenging to determine and identify the new SCs. In recent years, several types of indole/indazole amide-based SCs have been seized, but systematic research on these compounds remains limited. Therefore, developing rapid, sensitive, and accurate quantitative methods to determine new SCs are of great importance. Compared with high performance liquid chromatography (HPLC), ultra performance liquid chromatography (UPLC) shows higher resolution, better separation efficiency, and faster analysis speeds; thus, it can meet the demand for the quantitative analysis of indole/indazole amide-based SCs in seized materials. In this study, a UPLC method was developed for the simultaneous determination of five indole/indazole amide-based SCs, including N-(1-amino-3,3-dimethyl-1-oxobutan-2-yl)-1-butyl-1H-indazole-3-carboxamide (ADB-BUTINACA), methyl 2-(1-(4-fluorobutyl)-1H-indole-3-carboxamido)-3,3-dimethylbutanoate (4F-MDMB-BUTICA), N-(1-methoxy-3,3-dimethyl-1-oxobutan-2-yl)-1-(5-fluoropentyl)-1H-indole-3-carboxamide (5F-MDMB-PICA), methyl 3,3-dimethyl-2-(1-(pent-4-en-1-yl)-1H-indazole-3-carboxamido)butanoate (MDMB-4en-PINACA), and N-(adamantan-1-yl)-1-(4-fluorobutyl)-1H-indazole-3-carboxamide (4F-ABUTINACA) in electronic cigarette oil; these SCs have been detected with increasing frequency in seized materials in recent years. The main factors influencing the separation and detection performance of the proposed method, including the mobile phase, elution gradient, column temperature, and detection wavelength, were optimized. The proposed method successfully quantified the five SCs in electronic cigarette oil via the external standard method. The samples were extracted using methanol, and the target analytes were separated on a Waters ACQUITY UPLC CSH C18 column (100 mm×2.1 mm, 1.7 μm) at column temperature of 35 ℃ and flow rate of 0.3 mL/min. The injection volume was 1 μL. The mobile phase consisted of acetonitrile and ultrapure water, and gradient elution was employed. The detection wavelengths were 290 and 302 nm. The five SCs were completely separated within 10 min under optimized conditions and showed good linear relationships between 1-100 mg/L, with correlation coefficients (r2) of up to 0.9999. The limits of detection (LOD) and quantification (LOQ) were 0.2 and 0.6 mg/L, respectively. Precision was determined using standard solutions of the five SCs at mass concentrations of 1, 10, and 100 mg/L. The intra-day precision (n=6) was <1.5%, and the inter-day precision (n=6) was <2.2%. Accuracy was determined by spiking electronic cigarette oil with low (2 mg/L), moderate (10 mg/L), and high (50 mg/L) levels of the five SCs, with six replicates per determination. The recoveries of the five SCs were 95.5%-101.9%, and their relative standard deviations (RSDs, n=6) were 0.2%-1.5%, with accuracies ranging from -4.5% to 1.9%. The proposed method showed good performance when applied to the analysis of real samples. It is accurate, rapid, sensitive, and effective for the determination of five indole/indazole amide-based SCs in electronic cigarette oil. Thus, it satisfies the requirements for practical determination and provides a reference for the determination of SCs with similar structures by UPLC.

Identification and quantification of 10 indole/indazole carboxamide synthetic cannabinoids in 36 herbal blends by gas chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy.

Herbal blends containing synthetic cannabinoids have become popular alternatives to marijuana. The number of synthetic cannabinoids and speed of their emergence enable this group of compounds particularly challenging in terms of detection, monitoring, and responding. In this work, both gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (NMR) methods were developed for the identification and quantification of synthetic cannabinoids in herbal blends. Ten types of indole/indazole carboxamide synthetic cannabinoids, which showed different types of substitutions connected to nitrogen of the indole/indazole carboxamide, were detected in 36 herbal blends. The GC-MS fragmentation routes of indole/indazole carboxamide synthetic cannabinoids were discussed in detail for structure identification purpose. The concentration range of synthetic cannabinoid in 36 herbal blends was 1.9-50.6mg/g using GC-MS method, while 1.5-49.0mg/g by NMR method. Nicotine in herbal blends was quantified by NMR method without using reference material, and showed a variation of 5.3-44.7mg/g. For quantitative analysis, NMR method showed great advantage in the absence of reference material, while GC-MS method showed great merit for multiple-compound analysis when reference material was available. Therefore, for the quantitative analysis of new emerged synthetic cannabinoid in herbal blends, different methods could be chosen by considering whether reference material is available, as well as the number and types of synthetic cannabinoids detected in a single sample.

Isolation, detection and identification of synthetic cannabinoids in alternative formulations or dosage forms

Synthetic cannabinoids (SCs) first emerged on the market in head shops in the form of herbal mixtures, and quickly triggered an effort to reclassify them as controlled substances. More recently, with the increased popularity of vaping, new formulations of SCs have emerged. SCs are being dissolved or suspended in e-cigarette liquids and sold as “e-Liquids”. Furthermore, suppliers of SCs on impregnated paper sheets have been found on the black market. This could facilitate smuggling of SCs into prisons, exacerbating the problems SCs already create in prisons. The advent of such New Psychoactive Substance formulations/“new drug-delivery” systems pose a new issue for the detection and control of SCs. This work investigated various SC formulations, including e-cigarette liquids, bulk powder street samples and a set of blank A4 blotting papers believed to be impregnated with SCs to determine qualitatively their drug content. HPLC-DAD, HRMS, NMR and GC–MS provided conclusive structural identification and characterization of a number of different SCs in such alternative forms. Mass- and UV-triggered semipreparative LC-DAD-MS was successfully employed to purify SCs at a mg scale, facilitating their structural identification via NMR, and also paving the way for rapidly obtaining pure SC standards from commercial formulations. Finally, an authentic letter seized from a prison was also analysed and the two synthetic cannabinoids AMB-FUBINACA and MMB-CHMICA were detected. Our data further highlight recent reports about emerging trends which include applying SCs in alternative formulations (i.e. impregnated paper letters and e-liquids) and provide a database to help others identify these potentially toxic substances.

New Synthetic Cannabinoids Metabolism and Strategies to Best Identify Optimal Marker Metabolites.

Synthetic cannabinoids (SCs) were initially developed as pharmacological tools to probe the endocannabinoid system and as novel pharmacotherapies, but are now highly abused. This is a serious public health and social problem throughout the world and it is highly challenging to identify which SC was consumed by the drug abusers, a necessary step to tie adverse health effects to the new drug's toxicity. Two intrinsic properties complicate SC identification, their often rapid and extensive metabolism, and their generally high potency relative to the natural psychoactive Δ9-tetrahydrocannabinol in cannabis. Additional challenges are the lack of reference standards for the major urinary metabolites needed for forensic verification, and the sometimes differing illicit and licit status and, in some cases, identical metabolites produced by closely related SC pairs, i.e., JWH-018/AM-2201, THJ-018/THJ-2201, and BB-22/MDMB-CHMICA/ADB-CHMICA. We review current SC prevalence, establish the necessity for SC metabolism investigation and contrast the advantages and disadvantages of multiple metabolic approaches. The human hepatocyte incubation model for determining a new SC's metabolism is highly recommended after comparison to human liver microsomes incubation, in silico prediction, rat in vivo, zebrafish, and fungus Cunninghamella elegans models. We evaluate SC metabolic patterns, and devise a practical strategy to select optimal urinary marker metabolites for SCs. New SCs are incubated first with human hepatocytes and major metabolites are then identified by high-resolution mass spectrometry. Although initially difficult to obtain, authentic human urine samples following the specified SC exposure are hydrolyzed and analyzed by high-resolution mass spectrometry to verify identified major metabolites. Since some SCs produce the same major urinary metabolites, documentation of the specific SC consumed may require identification of the SC parent itself in either blood or oral fluid. An encouraging trend is the recent reduction in the number of new SC introduced per year. With global collaboration and communication, we can improve education of the public about the toxicity of new SC and our response to their introduction.

Simple screening procedure for 72 synthetic cannabinoids in whole blood by liquid chromatography\u2013tandem mass spectrometry

PurposeIn recent years, many synthetic cannabinoids (SCs) have appeared on the drug market. Despite the increasing number of SCs, there are few comprehensive screening methods for their detection in biological specimens. In this context, the purpose of this study was to develop a fast and simple liquid chromatography–tandem mass spectrometry screening procedure for detection and identification of SCs in whole blood.MethodsThe elaborated qualitative screening method allows the simultaneous detection and identification of 72 compounds from different chemical groups: naphthoylindoles, naphthoylindazoles, benzoylindoles, phenylacetylindoles, tetramethylcyclopropylindoles, indole-3-carboxylic acid esters, indole-3-carboxylic acid amides, indazole-3-carboxylic acid amides, and others. Whole-blood samples (0.2 mL) were precipitated with acetonitrile (0.6 mL). The separation was achieved with the gradient of the mobile phase composition (0.1% formic acid in acetonitrile and 0.1% formic acid in water) and the gradient of the flow rate (0.5–0.8 mL/min) in 16 min. Detection of all compounds was based on dynamic multiple reaction monitoring.ResultsMass spectrometer parameters for all compounds were presented. All of the compounds were well-separated by their retention times and/or transitions. The limits of detection (LODs) for 50 compounds were in the range 0.01–0.48 ng/mL.ConclusionsEstimated LODs make this assay suitable for the analysis of biological material. The procedure can be easily expanded for more substances, which is an indispensable advantage in the dynamically developing drug market. It can have wide application in various analytical forensic and clinical laboratories.

Screening for synthetic cannabinoids in human urine samples by LC-QToF-MS using software assisted metabolite identification for method updating

New synthetic cannabinoids (SC) frequently appear on the markets challenging toxicologists. Perceived as a legal alternative to cannabis a growing number of SCs is available. Consequently, analytical methods have to be adapted frequently to include newly emerging compounds. Since most SC are metabolized extensively metabolite identification prior to analyzing urine samples is inevitable. The presented study aimed at the development and implementation of a workflow allowing for the identification of the major in vitro phase I metabolites of new compounds by using dedicated software to mine analytical data of pooled human liver microsome (pHLM) incubations. The identified metabolites and their respective MSMS fragment information are utilized to update analytical methods for the reliable identification of SC uptake in human urine samples. The performed pHLM assay was according to an in-house protocol. The MS was operated in positive ESI with data-dependent MSMS fragmentation. MassMetaSite software (Molecular Discovery) was used to analyze HPLC-MSMS datasets of the incubations sampled at time points zero and one hour. Precursor and fragment information of the identified metabolites as well as their retention times were collected and added to a database used for LC-QToF-MS screening of urine samples for SC uptake. To allow for retrospective data evaluation authentic urine samples were analyzed in data independent broad-band-CID mode (bbCID). The highly potent and prevalent synthetic cannabinoid MDMB-CHMICA was chosen as a model compound. Analysis of the pHLM incubations of MDMB-CHMICA with MassMetaSite revealed 10 metabolites with at least two fragment masses each. Although in-depth manual data evaluation lead to the identification of 15 phase I metabolites, all main in vitro phase I metabolites were detected with the software. Authentic forensic case samples were screened after enzymatic cleavage of glucuronides and processed with the updated TASQ method. Despite varying relative abundances of the detected metabolites, the in vitro and in vivo data showed good agreement with respect to the chosen MDMB-CHMICA metabolites. Using MassMetaSite software and the described workflow proved to be a suitable procedure to identify metabolites and subsequently update analytical method to account for the rapid fluctuation of compounds among the group of new psychoactive substances (NPS). The here described approach can be helpful for updating screening methods with metabolite information. This is necessary whenever dealing with analytes that are extensively metabolized such as SCs. With other NPS the identification of metabolites along with the parent compound can serve as a plausibility check and may help in estimating the time of the last drug uptake. The use of dedicated software to help with metabolite identification is less time consuming thus providing a useful alternative for the rapid method update in routine forensic toxicology.

Identification and analytical characterization of six synthetic cannabinoids NNL-3, 5F-NPB-22-7N, 5F-AKB-48-7N, 5F-EDMB-PINACA, EMB-FUBINACA, and EG-018.

Clinical and forensic toxicology laboratories are continuously confronted by analytical challenges when dealing with the new psychoactive substances phenomenon. The number of synthetic cannabinoids, the chemical diversity, and the speed of emergence make this group of compounds particularly challenging in terms of detection, monitoring, and responding. Three indazole 7N positional isomer synthetic cannabinoids, two ethyl 2-amino-3-methylbutanoate-type synthetic cannabinoids, and one 9H-carbazole substituted synthetic cannabinoid were identified in seized materials. These six synthetic cannabinoid derivatives included: 1H-benzo[d] [1,2,3]triazol-1-yl 1-(5-fluoropentyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxylate (NNL-3, 1), quinolin-8-yl 1-(5-fluoropentyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxylate (5F-NPB-22-7N, 2), N-((1s,3s)-adamantan-1-yl)-1-(5-fluoropentyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (5F-AKB-48-7N, 3), ethyl 2-(1-(5-fluoropentyl)-1H-indazole-3-carboxamido)-3,3-dimethylbutanoate (5F-EDMB-PINACA, 4), ethyl 2-(1-(4-fluorobenzyl)-1H-indazole-3-carboxamido)-3-methylbutanoate (EMB-FUBINACA, 5), and naphthalen-1-yl(9-pentyl-9H-carbazol-3-yl)methanone (EG-018, 6). The identification was based on ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UHPLC-QTOF-MS), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance spectroscopy (NMR). The analytical characterization of these six synthetic cannabinoids was described, so as to assist forensic laboratories in identifying these compounds or other substances with similar structure in their case work. To our knowledge, no analytical data about the compounds 1-5 have appeared until now, making this the first report on these compounds. The GC-MS data of 6 has been reported, but this study added the LC-MS, NMR, and Fourier transform infrared (FTIR), data to render the analytical data collection process more complete. Copyright © 2017 John Wiley & Sons, Ltd.

Use of Synthetic Cannabinoids in Patients With Psychotic Disorders: Case Series

An increasing number of synthetic cannabinoids have become available on the black market in recent years, and health professionals have seen a corresponding increase in use of these compounds among patients with psychiatric disorders. Unfortunately, there is almost no research available in the literature on this topic, and what little exists is based on case reports of individuals without psychiatric disorders. Synthetic cannabinoids are functionally similar to, but structurally different from, delta-9-tetrahydrocannabinol, the active principle in cannabis, and are problematic for many reasons. The psychotropic action of synthetic cannabinoids in patients with schizophrenia is unpredictable, with very diverse clinical presentations. These drugs can be much more potent than delta-9-tetrahydrocannabinol, they are readily available and difficult to detect. The gold standard for identification of synthetic cannabinoids is gas chromatography with mass spectrometry, but even this is difficult because new formulations of these designer drugs are constantly emerging. In this manuscript, we provide an overview and discussion of synthetic cannabinoids and present four cases of patients with synthetic cannabinoid intoxication who were hospitalized in our intensive psychiatric unit at the time of intoxication. All patients had a history of schizophrenia and had been hospitalized several times previously. While hospitalized, they smoked an unknown substance brought in by a visitor, which was then confirmed using gas chromatography with mass spectrometry to be the synthetic cannabinoid AM-2201. Our patients experienced predominantly psychiatric adverse clinical effects. We observed the appearance of new psychotic phenomena, without exacerbation of their previously known psychotic symptoms, as well as the occurrence or marked worsening of mood and anxiety symptoms. Despite several similar reactions, and even though they ingested the same exact substance, the clinical picture differed markedly between individual patients. We assume that the acute effects of synthetic cannabinoids in patients with schizophrenia would be different from those in persons without psychotic disorders. The reasons for this difference could be the actual symptomatology of the presenting disorder, the impact of psychopharmacotherapy, individual patient differences and probably many, as yet unknown, factors. The long-term consequences of synthetic cannabinoid use on preexisting psychotic disorders are unclear.

Prevalence of synthetic cannabinoids in blood samples from Norwegian drivers suspected of impaired driving during a seven weeks period

From early year 2000 different herbal products containing synthetic cannabinoids (SC) have appeared on the drug market all over the world, and new substances are frequently introduced. The prevalence of SC use in different populations is however still mainly unknown, also in Norway. This information is difficult to obtain, but studies of drivers suspected of driving under the influence of drugs (DUID), might provide important information.The aim of this study was to assess the prevalence of SC in drivers suspected of being under the influence of drugs in Norway, and investigate if SCs impair driving performance.For two periods of three and four weeks all blood samples from drivers suspected of DUID in Norway were analyzed for the presence of 12 and 18 different SCs, respectively. A new ultra performance liquid chromatography tandem mass spectrometry method was developed.A total of 726 cases were analyzed during our study period, and SCs were detected in 16 cases (2.2%) in total. The mean age of these drivers was 29.6 years. High concentrations of other psychoactive drugs were detected in all the blood samples where a SC was found. AM-2201 and JWH-018 were the most frequently detected SCs, each found in five cases. In addition RSC-4, JWH-122, JWH-081 and JWH-250 were detected. None of the drivers had reported using SCs prior to driving.Despite the limited number of SCs investigated in this 7 week study period, a considerable percent of the cases were positive. Other psychoactive drugs of abuse were always found concomitant with the SCs, and the age of these drivers indicates that experienced drug users also ingest SCs. Since other drugs were found in all the samples, the psychomotor impairment caused by the SCs is difficult to estimate. Our study shows the importance of screening analyses of biological samples from different populations to assess the prevalence of drug use, since self-reporting might be encumbered with significant under-reporting.

Synthetic Cannabinoids: Crisis of The Decade

The Chinese emperor Shen-Nung was first to mention (2737 BC) the benefits of cannabis, noting its properties against malaria and rheumatism (1). Following the isolation of Δ9-tetrahydrocannabinoid (Δ9-THC) in 1964 (2) and discovery of cannabinoid receptors (CB1 and CB2) in the 1980s (3), a large number of cannabinoid receptor agonists have been synthesized for therapeutic purposes (3) while, on the other hand cannabis has become the most widely produced and consumed illicit substance worldwide (4). For many decades there have been few additions to the spectrum of drugs abused. However, in recent years, there has been a huge upsurge in novel psychoactive substances, also known as “legal highs”, “designer drugs”, “herbal highs” or “research chemicals”. Since 1997 more than 200 new psychoactive substances have been identified (5). In 1990s, Huffman et al. (6) synthesized naphthoylindoles, naphthoylpyrroles and related compounds with cannabinoid receptor agonist activity, which have become known as the “JWH compounds” and have become the major component of novel drugs containing synthetic cannabinoids (SCs) afterwards. Since 2004, SCs have become available on the market and they have become popular with those seeking a “legal high” (7). The SCs reported since 2008, belongs to different chemical groups named as naphthoylindoles, c y c lo hexy lp heno l s , t r i cyc l i c t e rpen o i ds , phenylacetylindoles, benzoylindoles, naphthoylpyrroles, naphthoylnaphthalenes, adamantoylindoles, quinones and cyclopropylindoles (8). Products containing SCs are commonly referred as “Spice” in Europe, “K2” in USA, whereas “Bonzai” or “Jamaika” in Turkey. These are “herbal smoking mixtures”, typically containing a number of different SCs that are sprayed onto the herbal constituents, which are then smoked by users, similar to cannabis (9,10). These drugs are widely marketed in Europe, the United States, and Japan, and easily accessible from the internet. “Herbal mixtures” of the “Spice”-type are labelled “not for human consumption” and are advertised e.g. as incense or plant growth regulator. These mixtures are declared to be purely herbal, but exhibit strong cannabimimetic effects after smoking because they have been adulterated with SC receptor agonists (10). Because of the lack of specific regulations and widespread use of products with unknown composition in terms of components and dosages, the growing consumption of SCs as designer drugs of abuse has become a significant trouble for public health institutions (11).

Prevalence of Synthetic Cannabinoids in U.S. Athletes: Initial Findings

A number of synthetic cannabinoids such as JWH-018 and JWH-073 have been incorporated into "spice" products. Despite having labels warning against human consumption, the products are smoked for their cannabinoid-like effects and the extent of their use by athletes has not been adequately described. Urine samples collected from 5,956 athletes were analyzed by high-performance liquid chromatography-tandem mass spectrometry for the presence of JWH-018, JWH-073, and their metabolites. Metabolites of JWH-018 and/or JWH-073 were detected in 4.5% of the samples. Metabolites of JWH-018 and JWH-073, only JWH-018, and only JWH-073 were detected in 50%, 49%, and approximately 1% of positive samples, respectively. In total, JWH-018 metabolites were detected in 99% (50% + 49%) and JWH-073 metabolites were detected in approximately 50% (49% + 1%) of the positive samples. Parent JWH-018, JWH-018-2-OH-indole, and JWH-018-4-OH-indole were not detected in any of the samples. All samples in which JWH-073 metabolites were detected contained JWH-073-N-butanoic acid. Parent JWH-073 and its N-(4-OH-butyl), 4-OH-indole, 5-OH-indole, and 7-OH-indole metabolites were not detected. Given the number of synthetic cannabinoids that have been synthesized, their limited regulation, and the prevalence of JWH-018 and JWH-073 metabolites detected in the athletes, these compounds should remain a priority for anti-doping programs.

Canabinoides sintéticos: drogas de abuso emergentes

CONTEXTO: A Cannabis sativa (cannabis, maconha, marijuana) é uma droga de abuso com efeitos psicoativos e potencial terapêutico bastante conhecidos. Um grande número de canabinoides foi sintetizado na tentativa de excluir ou minimizar os efeitos psicotrópicos e mantendo as aplicações terapêuticas. Os canabinoides sintéticos representam o mais recente avanço das designer drugs. OBJETIVOS: Este trabalho busca trazer informações sobre a utilização de canabinoides sintéticos como drogas de abuso emergentes, principalmente sob a forma de spice ou herbal blends. MÉTODOS: Foi realizada uma pesquisa bibliográfica na base de dados PubMed utilizando os descritores de assunto "synthetic cannabinoids", "spice", "legal highs", "herbal blends", "psychosis cannabis" e suas combinações. RESULTADOS: Foram encontrados canabinoides sintéticos nos produtos Spice, confirmando que os incensos herbais realmente surgiram como drogas de abuso emergentes. CONCLUSÃO: Não está claro se a utilização de canabinoides sintéticos e de Spice é uma moda passageira ou se irá se estabelecer em nossa sociedade. No entanto, o fenômeno das designer drugs, especialmente canabinoides sintéticos, representa um desafio presente para o sistemas de saúde no mundo globalizado, tornando-se necessária a pesquisa clínica e forense desses produtos.