Analytical characterization of four new ortho‐methoxybenzylated amphetamine‐type designer drugs

In a seizure of German custom authorities four N‐(ortho‐methoxybenzyl)amines with amphetamine partial structure were obtained as pure compounds: N‐(ortho‐methoxybenzyl)‐3,4‐dimethoxyamphetamine (3,4‐DMA‐NBOMe ( 1 )), N‐(ortho‐methoxybenzyl)‐4‐ethylamphetamine (4‐EA‐NBOMe ( 2 )), N‐(ortho‐methoxybenzyl)‐4‐methylmethamphetamine (4‐MMA‐NBOMe ( 3 )), and N‐(ortho‐methoxybenzyl)‐5‐(2‐aminopropyl)benzofuran (5‐APB‐NBOMe ( 4 )). The compounds have been detected in Germany for the first time and no analytical data had been previously published. Mass spectrometric (MS), infrared (IR) spectroscopic, and nuclear magnetic resonance (NMR) spectroscopic data are presented. Copyright © 2015 John Wiley & Sons, Ltd.     

In vitro characterization of potential CYP‐ and UGT‐derived metabolites of the psychoactive drug 25B‐NBOMe using LC‐high resolution MS

One of the main challenges posed by the emergence of new psychoactive substances is their identification in human biological samples. Trying to detect the parent drug could lead to false‐negative results when the delay between consumption and sampling has been too long. The identification of their metabolites could then improve their detection window in biological matrices. Oxidative metabolism by cytochromes P450 and glucuronidation are two major detoxification pathways in humans. In order to characterize possible CYP‐ and UGT‐dependent metabolites of the 2‐(4‐bromo‐2,5‐dimethoxy‐phenyl)‐N‐[(2‐methoxyphenyl)methyl]ethanamine (25B‐NBOMe), a synthetic psychoactive drug, analyses of human liver microsome (HLM) incubates were performed using an ultra‐high performance liquid chromatography system coupled with a quadrupole‐time of flight mass spectrometry detector (UHPLC‐Q‐TOF/MS). On‐line analyses were performed using a Waters OASIS HLB column (30 x 2.1 mm, 20 µm) for the automatic sample loading and a Waters ACQUITY HSS C18 column (150 x 2 mm, 1.8 µm) for the chromatographic separation. Twenty‐one metabolites, consisting of 12 CYP‐derived and 9 UGT‐derived metabolites, were identified. O‐Desmethyl metabolites were the most abundant compounds after the phase I process, which appears to be in accordance with data from previously published NBOMe‐intoxication case reports. Although other important metabolic transformations, such as sulfation, acetylation, methylation or glutathione conjugation, were not studied and artefactual metabolites might have been produced during the HLM incubation process, the record of all the metabolite MS spectra in our library should enable us to characterize relevant metabolites of 25B‐NBOMe and allow us to detect 25B‐MBOMe users. Copyright © 2015 John Wiley & Sons, Ltd.     

Identification of five substituted phenethylamine derivatives 5‐MAPDB, 5‐AEDB,MDMA methylene homolog, 6‐Br‐MDMA,and 5‐APB‐NBOMe

This paper reports analytical properties of five substituted phenethylamine derivatives seized from a clandestine laboratory. These five derivatives include 5‐(2‐methylaminopropyl)‐2,3‐dihydrobenzofuran (5‐MAPDB, 1 ), 5‐(2‐aminoethyl)‐2,3‐dihydrobenzofuran (5‐AEDB, 2 ), N ,2‐dimethyl‐3‐(3,4‐methylenedioxyphenyl)propan‐1‐amine (MDMA methylene homolog, 3 ), 6‐bromo‐3,4‐methylenedioxymethamphetamine (6‐Br‐MDMA, 4 ), and 1‐(benzofuran‐5‐yl)‐N ‐(2‐methoxybenzyl)propan‐2‐amine (5‐APB‐NBOMe, 5 ). These compounds were identified by liquid chromatography‐quadrupole time‐of‐flight mass spectrometry (LC‐QTOF‐MS), gas chromatography‐mass spectrometry (GC‐MS), and nuclear magnetic resonance spectroscopy (NMR). No analytical properties about compounds 1‐4 have appeared until now, making this the first report on these compounds. Copyright © 2016 John Wiley & Sons, Ltd.     

25C‐NBOMe and 25I‐NBOMe metabolite studies in human hepatocytes,in vivo mouse and human urine with high‐resolution mass spectrometry

25C‐NBOMe and 25I‐NBOMe are potent hallucinogenic drugs that recently emerged as new psychoactive substances. To date, a few metabolism studies were conducted for 25I‐NBOMe, whereas 25C‐NBOMe metabolism data are scarce. Therefore, we investigated the metabolic profile of these compounds in human hepatocytes, an in vivo mouse model and authentic human urine samples from forensic cases. Cryopreserved human hepatocytes were incubated for 3 h with 10 μM 25C‐NBOMe and 25I‐NBOMe; samples were analyzed by liquid chromatography high‐resolution mass spectrometry (LC‐HRMS) on an Accucore C18 column with a Thermo QExactive; data analysis was performed with Compound Discoverer software (Thermo Scientific). Mice were administered 1.0 mg drug/kg body weight intraperitoneally, urine was collected for 24 h and analyzed (with or without hydrolysis) by LC‐HRMS on an Acquity HSS T3 column with an Agilent 6550 QTOF; data were analyzed manually and with WebMetabase software (Molecular Discovery). Human urine samples were analyzed similarly. In vitro and in vivo results matched well. 25C‐NBOMe and 25I‐NBOMe were predominantly metabolized by O‐demethylation, followed by O‐di‐demethylation and hydroxylation. All methoxy groups could be demethylated; hydroxylation preferably occurred at the NBOMe ring. Phase I metabolites were extensively conjugated in human urine with glucuronic acid and sulfate. Based on these data and a comparison with synthesized reference standards for potential metabolites, specific and abundant 25C‐NBOMe urine targets are 5’‐desmethyl 25C‐NBOMe, 25C‐NBOMe and 5‐hydroxy 25C‐NBOMe, and for 25I‐NBOMe 2’ and 5’‐desmethyl 25I‐NBOMe and hydroxy 25I‐NBOMe. These data will help clinical and forensic laboratories to develop analytical methods and to interpret results. Copyright © 2016 John Wiley & Sons, Ltd.     

Characterization of the hepatic cytochrome P450 enzymes involved in the metabolism of 25I‐NBOMe and 25I‐NBOH

The dimethoxyphenyl‐N‐((2‐methoxyphenyl)methyl)ethanamine (NBOMe) compounds are potent serotonin 5‐HT2A receptor agonists and have recently been subject to recreational use due to their hallucinogenic effects. Use of NBOMe compounds has been known since 2011, and several non‐fatal and fatal intoxication cases have been reported in the scientific literature. The aim of this study was to determine the importance of the different cytochrome P450 enzymes (CYP) involved in the metabolism of 2‐(4‐iodo‐2,5‐dimethoxyphenyl)‐N‐(2methoxybenzyl)ethanamine (25I‐NBOMe) and 2‐[[2‐(4‐iodo‐2,5dimethoxyphenyl)ethylamino]methyl]phenol (25I‐NBOH) and to characterize the metabolites. The following approaches were used to identify the main enzymes involved in primary metabolism: incubation with a panel of CYP and monoamine oxidase (MAO) enzymes and incubation in pooled human liver microsomes (HLM) with and without specific CYP chemical inhibitors. The study was further substantiated by an evaluation of 25I‐NBOMe and 25I‐NBOH metabolism in single donor HLM. The metabolism pathways of 25I‐NBOMe and 25I‐NBOH were NADPHdependent with intrinsic clearance values of (CLint) of 70.1 and 118.7 mL/min/kg, respectively. The biotransformations included hydroxylation, O‐demethylation, N‐dealkylation, dehydrogenation, and combinations thereof. The most abundant metabolites were all identified by retention time and spectrum matching with synthesized reference standards. The major CYP enzymes involved in the metabolism of 25I‐NBOMe and 25INBOH were identified as CYP3A4 and CYP2D6, respectively. The compound 25I‐NBOH was also liable to direct glucuronidation, which may diminish the impact of CYP2D6 genetic polymorphism. Users of 25I‐NBOMe may be subject to drug‐drug interactions (DDI) if 25I‐NBOMe is taken with a strong CYP3A4 inhibitor. Copyright © 2016 John Wiley & Sons, Ltd.     

Metabolism of the tryptamine‐derived new psychoactive substances 5‐MeO‐2‐Me‐DALT, 5‐MeO‐2‐Me‐ALCHT,and 5‐MeO‐2‐Me‐DIPT and their detectability in urine studied by GC–MS,LC–MSn,and LC‐HR‐MS/MS

Many N,N‐dialkylated tryptamines show psychoactive properties and were encountered as new psychoactive substances. The aims of the presented work were to study the phase I and II metabolism and the detectability in standard urine screening approaches (SUSA) of 5‐methoxy‐2‐methyl‐N,N‐diallyltryptamine (5‐MeO‐2‐Me‐DALT), 5‐methoxy‐2‐methyl‐N‐allyl‐N‐cyclohexyltryptamine (5‐MeO‐2‐Me‐ALCHT), and 5‐methoxy‐2‐methyl‐N,N‐diisopropyltryptamine (5‐MeO‐2‐Me‐DIPT) using gas chromatography–mass spectrometry (GC–MS), liquid chromatography coupled with multistage accurate mass spectrometry (LC–MSⁿ), and liquid chromatography‐high‐resolution tandem mass spectrometry (LC‐HR‐MS/MS). For metabolism studies, urine was collected over a 24 h period after administration of the compounds to male Wistar rats at 20 mg/kg body weight (BW). Phase I and II metabolites were identified after urine precipitation with acetonitrile by LC‐HR‐MS/MS. 5‐MeO‐2‐Me‐DALT (24 phase I and 12 phase II metabolites), 5‐MeO‐2‐Me‐ALCHT (24 phase I and 14 phase II metabolites), and 5‐MeO‐2‐Me‐DIPT (20 phase I and 11 phase II metabolites) were mainly metabolized by O‐demethylation, hydroxylation, N‐dealkylation, and combinations of them as well as by glucuronidation and sulfation of phase I metabolites. Incubations with mixtures of pooled human liver microsomes and cytosols (pHLM and pHLC) confirmed that the main metabolic reactions in humans and rats might be identical. Furthermore, initial CYP activity screenings revealed that CYP1A2, CYP2C19, CYP2D6, and CYP3A4 were involved in hydroxylation, CYP2C19 and CYP2D6 in O‐demethylation, and CYP2C19, CYP2D6, and CYP3A4 in N‐dealkylation. For SUSAs, GC–MS, LC‐MSⁿ, and LC‐HR‐MS/MS were applied to rat urine samples after 1 or 0.1 mg/kg BW doses, respectively. In contrast to the GC–MS SUSA, both LC–MS SUSAs were able to detect an intake of 5‐MeO‐2‐Me‐ALCHT and 5‐MeO‐2‐Me‐DIPT via their metabolites following 1 mg/kg BW administrations and 5‐MeO‐2‐Me‐DALT following 0.1 mg/kg BW dosage. Copyright © 2017 John Wiley & Sons, Ltd.     

Synthesis and determination of analytical characteristics and differentiation of positional isomers in the series of N‐(2‐methoxybenzyl)‐2‐(dimethoxyphenyl)ethanamine using chromatography–mass spectrometry

N‐(2‐Methoxybenzyl)‐2,5‐dimethoxyphenethylamines (NBOMes) are synthetic phenethylamine derivatives emerging on the global drug market and reported to be associated with untoward effects in people who use drugs. Its action involves agonism at serotonin 5‐HT_2A receptors, affecting cognitive and behavioral processes. However, certain isomers of NBOMes may not show any psychoactive effects. They are not controlled by legislation and can be tested as pharmaceutical drugs. This study deals with the differentiation among positional isomers of 25H‐NBOMe differing in the position of the two methoxy groups in the phenylethyl moiety of the molecule, using chromatography–mass spectrometry methods. The gas chromatography analysis showed that the isothermal mode was more efficient than the usually applied temperature‐programming mode for the separation of the mentioned isomers. Electron ionization mass spectra of 25H‐NBOMe isomers were highly similar, often resulting in a high probability of erroneous identification. However, mass spectra of their trifluoroacetyl or pentafluoropropanoyl derivatives were easily identified as they contained fragments with many significant differences. The proposed analysis using liquid chromatography–tandem mass spectrometry could distinguish the isomers of 25H‐NBOMe without the need for any derivatization.     

Identification and characterization of 2,5‐dimethoxy‐3,4‐dimethyl‐β‐phenethylamine (2C‐G) – A new designer drug

This study presents and discusses the mass spectrometric, infrared spectroscopic and nuclear magnetic resonance spectroscopic data of 2,5‐dimethoxy‐3,4‐dimethyl‐β‐phenethylamine (2C‐G), a new designer drug. A powder sample containing 2C‐G was seized in Poland in 2011. The paper focuses on a comparison of the analytical features of 2C‐G and other members of the 2C‐series, in order to assess the possibility of unequivocal identification. The occurrence of intense peak at m/z = 178 and different intensities of the ions at m/z = 165 and 180 in the gas chromatography‐electron impact‐mass spectrometry (GC‐EI/MS) spectrum of 2C‐G made it possible to distinguish it from 2C‐E. Differences in relative intensities of the ions at m/z = 192, 179 and 177 were observed for GC‐EI/MS spectra of TFAA derivatives of 2C‐G and 2C‐E. An identical set of ions was recorded for these substances using the liquid chromatography‐electrospray ionization/quadrupole time of flight mass spectrometry (LC‐ESI/QTOFMS) method in both MS and tandem mass spectrometry (MS/MS) mode, but the distinction was possible based on differences in the ion intensities at m/z = 193.1223 and 178.0988. The Fourier transform infrared (FTIR) spectrum of 2C‐G was significantly different from other members of the 2C‐series, with a characteristic doublets at 993–1014 cm^‐1 and 1099–1124 cm^‐1, and the ratio of bands at higher wavenumbers. Final elucidation of the structure of 2C‐G was carried out by ¹H and ¹³C NMR spectroscopy. The study indicated that the marketing of analogues of controlled substances poses a real analytical challenge for forensic laboratories, and the application of sophisticated methods is often required for unequivocal identification of a new substance. Copyright © 2012 John Wiley & Sons, Ltd.     

Characterization of hallucinogenic phenethylamines using high‐resolution mass spectrometry for non‐targeted screening purposes

Hallucinogenic phenethylamines such as 2,5‐dimethoxyphenethylamines (2C–X) and their N ‐(2‐methoxybenzyl) derivatives (25X–NBOMe) have seen an increase in novel analogues in recent years. These rapidly changing analogues make it difficult for laboratories to rely on traditional targeted screening methods to detect unknown new psychoactive substances (NPS). In this study, twelve 2C–X, six 2,5‐dimethoxyamphetamines (DOX), and fourteen 25X–NBOMe derivatives, including two deuterated derivatives (2C–B‐d ₆ and 25I–NBOMe‐d ₉), were analyzed using ultra‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry (UPLC‐QTOF‐MS). Collision‐induced dissociation (CID) experiments were performed using collision energies set at 10, 20, and 40 eV. For 2C–X and DOX derivatives, common losses were observed including neutral and radical losses such as NH₃ (17.0265 Da), •CH₆N (32.0500 Da), C₂H₇N (45.0578 Da) and C₂H₉N (47.0735 Da). 2C–X derivatives displayed common product ions at m/z 164.0837 ([C₁₀H₁₂O₂]^+•), 149.0603 ([C₉H₉O₂]⁺), and 134.0732 ([C₉H₁₀O]^+•) while DOX derivatives had common product ions at m/z 178.0994 ([C₁₁H₁₄O₂]^+•), 163.0754 ([C₁₀H₁₁O₂]⁺), 147.0804 ([C₁₀H₁₁O]⁺), and 135.0810 ([C₉H₁₁O]⁺). 25X–NBOMe had characteristic product ions at m/z 121.0654 ([C₈H₉O]⁺) and 91.0548 ([C₇H₇]⁺) with minor common losses corresponding to 2‐methylanisole (C₈H₁₀O, 122.0732 Da), 2‐methoxybenzylamine (C₈H₁₁NO, 137.0847 Da), and •C₉H₁₄NO (152.1074 Da). Novel analogues of the selected classes can be detected by applying neutral loss filters (NLFs) and extracting the common product ions. Copyright © 2017 John Wiley & Sons, Ltd.     

Identification and analytical characterization of four synthetic cathinone derivatives iso‐4‐BMC, β‐TH‐naphyrone,mexedrone, and 4‐MDMC

New psychoactive substances (NPS) have gained much popularity on the global market over the last number of years. The synthetic cathinone family is one of the most prominent groups and this paper reports on the analytical properties of four synthetic cathinone derivatives: ( 1 ) 1‐(4‐bromophenyl)‐1‐(methylamino)propan‐2‐one (iso‐4‐BMC or iso‐brephedrone), ( 2 ) 2‐(pyrrolidin‐1‐yl)‐1‐(5,6,7,8‐tetrahydronaphthalen‐2‐yl)pentan‐1‐one (β ‐TH‐naphyrone), ( 3 ) 3‐methoxy‐2‐(methylamino)‐1‐(4‐methylphenyl)propan‐1‐one (mexedrone), and ( 4 ) 2‐(dimethylamino)‐1‐(4‐methylphenyl)propan‐1‐one (4‐MDMC). These identifications were based on liquid chromatography‐quadrupole time‐of‐flight‐mass spectrometry (LC‐QTOF‐MS), gas chromatography‐mass spectrometry (GC‐MS) and nuclear magnetic resonance (NMR) spectroscopy. To our knowledge, no chemical or pharmacological data about compounds 1–3 have appeared until now, making this the first report on these compounds. The Raman and GC‐MS data of 4 have been reported, but this study added the LC‐MS and NMR data for additional characterization. Copyright © 2016 John Wiley & Sons, Ltd.     

Phenethylamine‐derived new psychoactive substances 2C‐E‐FLY, 2C‐EF‐FLY,and 2C‐T‐7‐FLY: Investigations on their metabolic fate including isoenzyme activities and their toxicological detectability in urine screenings

Psychoactive substances of the 2C‐series are phenethylamine‐based designer drugs that can induce psychostimulant and hallucinogenic effects. The so‐called 2C‐FLY series contains rigidified methoxy groups integrated in a 2,3,6,7‐tetrahydrobenzo[1,2‐b:4,5‐b']difuran core. The aim of the presented work was to investigate the in vivo and in vitro metabolic fate including isoenzyme activities and toxicological detectability of the three new psychoactive substances (NPS) 2C‐E‐FLY, 2C‐EF‐FLY, and 2C‐T‐7‐FLY to allow clinical and forensic toxicologists the identification of these novel compounds. Rat urine, after oral administration, and pooled human liver S9 fraction (pS9) incubations were analyzed by liquid chromatography?high‐resolution tandem mass spectrometry (LC?HRMS/MS). By performing activity screenings, the human isoenzymes involved were identified and toxicological detectability in rat urine investigated using standard urine screening approaches (SUSAs) based on gas chromatography (GC)?MS, LC?MSⁿ, and LC?HRMS/MS. In total, 32 metabolites were tentatively identified. Main metabolic steps consisted of hydroxylation and N‐acetylation. Phase I metabolic reactions were catalyzed by CYP2D6, 3A4, and FMO3 and N‐acetylation by NAT1 and NAT2. Methoxyamine was used as a trapping agent for detection of the deaminated metabolite formed by MAO‐A and B. Interindividual differences in the metabolism of the 2C‐FLY drugs could be caused by polymorphisms of enzymes involved or drug–drug interactions. All three SUSAs were shown to be suitable to detect an intake of these NPS but common metabolites of 2C‐E‐FLY and 2C‐EF‐FLY have to be considered during interpretation of analytical findings.     

Liquid chromatography‐quadrupole‐time‐of‐flight mass spectrometry screening procedure for urine samples in forensic casework compared to gas chromatography‐mass spectrometry

This work represents the development, validation, and application of a liquid chromatography‐quadrupole‐time‐of‐flight mass spectrometry (LC‐QTOF‐MS) screening method for the detection of pharmaceutical substances and illicit drugs (acidic, basic, and neutral organic drugs) in urine samples. Time‐of‐flight mass spectrometry was performed using an LC‐Triple TOF 5600 system with electrospray ionization operated in both positive and negative mode, respectively. The limits of detection (LODs), determined for 34 substances, were < 10 ng/mL for 91% of the compounds. The limits of quantitation (LOQs) were < 20 ng/mL for 91% of the substances. The identification of the compounds was based on exact mass (< ± 5 ppm), retention time (<2%) if available, isotopic pattern fit (<10%) and library hit (>70%). These four parameters served as identification criteria and are discussed according to their role in identifying compounds even without reference substances. In routine casework, two in‐house XIC (extracted ion chromatogram) lists, consisting of 456 protonated and 26 deprotonated compounds were used and retention times for 365 compounds were available. Compared to the results found with the established gas chromatography‐mass spectrometry (GC‐MS) procedure, the findings with the LC‐QTOF‐MS screening method showed a good comparability. Results that were not detected by LC‐QTOF‐MS because of a missing entry in the targeted XIC list could retrospectively be confirmed by simply entering the elemental formula of the relevant substance into the software and reprocessing the sample. LC‐QTOF‐MS offers an attractive technique for the fast and specific identification of illicit drugs and toxic compounds in urine samples. Copyright © 2016 John Wiley & Sons, Ltd.     

The chemistry and pharmacology of synthetic cannabinoid SDB‐006 and its regioisomeric fluorinated and methoxylated analogs

Synthetic cannabinoids are the largest and most structurally diverse class of new psychoactive substances, with manufacturers often using isomerism to evade detection and circumvent legal restriction. The regioisomeric methoxy‐ and fluorine‐substituted analogs of SDB‐006 (N‐benzyl‐1‐pentyl‐1H‐indole‐3‐carboxamide) were synthesized and could not be differentiated by gas chromatography–mass spectrometry (GC–MS), but were distinguishable by liquid chromatography–quadrupole time‐of‐flight–MS (LC–QTOF–MS). In a fluorescence‐based plate reader membrane potential assay, SDB‐006 acted as a potent agonist at human cannabinoid receptors (CB₁ EC₅₀ = 19 nM). All methoxy‐ and fluorine‐substituted analogs showed reduced potency compared to SDB‐006, although the 2‐fluorinated analog (EC₅₀ = 166 nM) was comparable to known synthetic cannabinoid RCS‐4 (EC₅₀ = 146 nM). Using biotelemetry in rats, SDB‐006 and RCS‐4 evoked comparable reduction in body temperature (~0.7°C at a dose of 10 mg/kg), suggesting lower potency than the recent synthetic cannabinoid AB‐CHMINACA (>2°C, 3 mg/kg).     

Synthesis of [13C6]‐labelled phenethylamine derivatives for drug quantification in biological samples

The availability of high‐quality ¹³C‐labelled internal standards will improve accurate quantification of narcotics and drugs in biological samples. Thus, the synthesis of 10 [¹³C₆]‐labelled phenethylamine derivatives, namely amphetamine, methamphetamine, 3,4‐methylenedioxyamphetamine, 3,4‐methylenedioxymethamphetamine, 3,4‐methylenedioxy‐N‐ethylamphetamine, 4‐methoxyamphetamine, 4‐methoxymethamphetamine, 3,5‐dimethoxyphenethylamine 4‐bromo‐2,5‐dimethoxyphenethylamine and 2,5‐dimethoxy‐4‐iodophenethylamine, have been undertaken. [¹³C₆]‐Phenol proved to be an excellent starting material for making ¹³C‐labelled narcotic substances in the phenethylamine class, and a developed Stille‐type coupling enabled an efficient synthesis of the 3,4‐methylenedioxy and 4‐methoxy derivatives. The pros and cons of alternative routes and transformations are also discussed. The [¹³C₆]‐labelled compounds are intended for use as internal standards in forensic analysis, health sciences and metabolomics studies by gas chromatography‐mass spectrometry and liquid chromatography‐tandem mass spectrometry. Copyright © 2014 John Wiley & Sons, Ltd.     

Synthesis of deuterium labeled phenethylamine derivatives

The synthesis of a series of five deuterium labeled phenethylamine derivatives, 4‐bromo‐2,5‐[²H₆]‐dimethoxyphenethylamine (2C‐B), 4‐chloro‐2,5‐[²H₆]‐dimethoxyphenethylamine (2C‐C), 2,5‐[²H₆]‐dimethoxy‐4‐iodophenethylamine (2C‐I), 2,5‐[²H₆]‐dimethoxy‐4‐ethylthiophenethylamine (2C‐T‐2) and 2,5‐[²H₆]‐dimethoxy‐4‐n‐propylthiophenethylamine (2C‐T‐7) from 1,4‐[²H₆]‐dimethoxybenzene is described. The isotopically labeled compounds are used as internal standards in gas chromatography‐mass spectrometry (GC‐MS) assays. Copyright © 2006 John Wiley & Sons, Ltd.     

Synthesis and Biological Activity of Some 1,3‐Dihydro‐2H‐3‐benzazepin‐2‐ones with a Piperazine Moiety as Bradycardic Agents

A series of 1,3‐dihydro‐2H‐3‐benzazepin‐2‐ones with a piperazine moiety were designed and synthesized by treating the common intermediate of 1,3‐dihydro‐7,8‐dimethoxy‐3‐[3‐(1‐piperazinyl)propyl]‐2H‐3‐benzazepin‐2‐ones with a variety of N‐aryl‐2‐chloroacetamides and acyl chlorides. Their structures have been characterized by ¹H‐NMR, MS, and elemental analysis. The title compounds were evaluated for their bradycardic activity in vitro. Most of the synthesized compounds exhibited some vasorelaxant activity and heart‐rate‐reducing activity with bradycardic potency.     

The identification and analytical characterization of 2,2′‐difluorofentanyl

New psychoactive substances (NPS) have expanded their distribution and become widely available in the global market in recent years. The illicit use of fentanyl and its analogs has become an important worldwide concern linked to their high potency and risk of fatal overdose. This study describes the analytical characterization of a new fentanyl derivative N‐(1‐(2‐fluorophenethyl)‐4‐piperidinyl)‐N‐(2‐fluorophenyl)propionamide (2,2′‐difluorofentanyl). Identification was based on ultra‐high‐performance liquid chromatography–quadrupole time‐of‐flight–mass spectrometry (UHPLC–QTOF–MS), gas chromatography–mass spectrometry (GC–MS), Fourier transform infrared (FTIR) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. To our knowledge, this study is the first to report on analytical data for this compound. The most abundant fragment ion in the electrospray ionization (ESI) mass spectrum under collision‐induced dissociation (CID) mode was formed by the cleavage between the piperidine ring and the N‐phenyl‐amide moiety of the protonated molecule. Two diagnostic ions in the electron ionization (EI) mass spectrum were formed by the loss of a tropylium ion (M‐91), and by the degradation of the piperidine ring and dissociate of the COC₂H₅ moiety altogether, respectively.     

Syntheses and analytical characterizations of N‐alkyl‐arylcyclohexylamines

The rise in new psychoactive substances that are available as 'research chemicals’ (RCs) remains a significant forensic and legislative challenge. A number of arylcyclohexylamines have attracted attention as RCs and continue to be encountered, including 3‐MeO‐PCP, 3‐MeO‐PCE and 3‐MeO‐PCPr. These compounds are commonly perceived as ketamine‐like dissociative substances and are believed to act predominantly via antagonism of the N‐methyl‐D‐aspartate (NMDA) receptor. To aid in the identification of newly emerging substances of abuse, the current studies were performed. The syntheses of fifteen N‐alkyl‐arylcyclohexylamines are described. Analytical characterizations were performed via gas chromatography and high performance liquid chromatography coupled to multiple forms of mass spectrometry as well as nuclear magnetic resonance spectroscopy, ultraviolet diode array detection and infrared spectroscopy. The series consisted of the N‐alkyl derivatives (N‐methyl, N‐ethyl, N‐propyl) of phenyl‐substituted and isomeric 2‐, 3‐ and 4‐methoxy phenylcyclohexylamines, as well as the N‐alkyl derivatives obtained from 3‐methylphenyl and 2‐thienyl moieties. In addition to the presentation of a range of previously unreported data, it was also found that positional isomers of aryl methoxyl‐substituted arylcyclohexylamines were readily distinguishable under a variety of analytical conditions. Copyright © 2015 John Wiley & Sons, Ltd.     

Synthesis and Positive Inotropic Evaluation of 2‐(4‐(4‐Substituted benzyloxy)‐3‐methoxybenzyl)‐1,4‐diazepan‐1‐yl)‐N‐(4,5‐dihydro‐1‐methyl[1,2,4]triazolo[4,3‐a]quinolin‐7‐yl)‐acetamides

In an attempt to search for more potent positive inotropic agents, a series of 2‐(4‐(4‐substituted benzyloxy)‐3‐methoxybenzyl)‐1,4‐diazepan‐1‐yl)‐N‐(4,5‐dihydro‐1‐methyl[1,2,4]triazolo[4,3‐a]quinolin‐7‐yl)acetamides was synthesized and their positive inotropic activities were evaluated by measuring left atrium stroke volume on isolated rabbit‐heart preparations. Several compounds showed favorable activity compared with the standard drug Milrinone among which 2‐(4‐(4‐(2‐chlorobenzyloxy)‐3‐methoxybenzyl)‐1,4‐diazepan‐1‐yl)‐N‐(4,5‐dihydro‐1‐methyl‐[1,2,4]triazolo[4,3‐a]quinolin‐7‐yl)acetamide 6e was found to have the most desirable potency with the 6.79 ± 0.18% increased stroke volume (Milrinone: 1.67 ± 0.64%) at a concentration of 1×10^–5 M in our in‐vitro study. The chronotropic effects of those compounds having inotropic effects were also evaluated in this work.     

Identification of three cannabimimetic indazole and pyrazole derivatives,APINACA 2H‐indazole analogue,AMPPPCA, and 5F‐AMPPPCA

This paper reports analytical properties of three cannabimimetic indazole and pyrazole derivatives seized from a clandestine laboratory. These three new synthetic cannabinoids include N ‐(1‐adamantyl)‐2‐pentyl‐2H ‐indazole‐3‐carboxamide (APINACA 2H ‐indazole analogue, 1 ), N ‐(1‐adamantyl)‐4‐methyl‐1‐pentyl‐5‐phenyl‐1H ‐pyrazole‐3‐carboxamide (AMPPPCA, 2 ), and N ‐(1‐adamantyl)‐1‐(5‐fluoropentyl)‐4‐methyl‐5‐phenyl‐1H ‐pyrazole‐3‐carboxamide (5F‐AMPPPCA, 3 ). These compounds were identified by liquid chromatography‐quadrupole time‐of‐flight‐mass spectrometry (LC‐QTOF‐MS), gas chromatography‐time‐of‐flight‐mass spectrometry (GC‐TOF‐MS), and nuclear magnetic resonance (NMR) spectroscopy. No analytical properties and pharmacological activities about compounds 1–3 have appeared until now, making this the first report on these compounds. Copyright © 2016 John Wiley & Sons, Ltd.