Related Designer Drugs Research Articles

Detection and identification of amphetamine-type stimulants and analogs via recognition-enabled “chromatographic” 19F NMR

The abuse of amphetamine (AMPH)-type stimulants (ATS) is a worldwide problem, and there is a need for reliable analytical methods to detect amphetamines and related designer drugs in unknown samples. Nuclear magnetic resonance (NMR) is an indispensable tool for structural elucidation and noninvasive analysis. Herein, we applied recognition-enabled “chromatographic” (REC) 19F NMR for the detection and quantitative analysis of ATS. This method allows for the detection and differentiation of up to 20 ATS and structurally similar analogs in complex mixtures, where the 19F NMR signals are precisely matched with those obtained in the analysis of each component independently. Selecting AMPH as a model analyte, we also illustrate the capability of our approach to quantitatively detect ATS. Further experiments showed that AMPH can be detected at a concentration of 1 μM in artificial urine. These results demonstrate that REC 19F NMR is an efficient and robust strategy for the determination of ATS and related derivatives.

Baths salts, spice, and related designer drugs: the science behind the headlines.

The abuse of synthetic psychoactive substances known as "designer drugs," or "new psychoactive substances" (NPS), is increasing at an alarming rate. NPS are purchased as alternatives to traditional illicit drugs of abuse and are manufactured to circumvent laws regulating the sale and use of controlled substances. Synthetic cathinones (i.e., "bath salts") and synthetic cannabinoids (i.e., "spice") are two types of NPS that have received substantial media attention. Although low recreational doses of bath salts or spice compounds can produce desirable effects, high doses or chronic exposure often leads to dangerous medical consequences, including psychosis, violent behaviors, tachycardia, hyperthermia, and even death. Despite the popularity of NPS, there is a paucity of scientific data about these drugs. Here we provide a brief up-to-date review describing the mechanisms of action and neurobiological effects of synthetic cathinones and cannabinoids.

P54 Capillary electrophoresis – mass spectrometry for the forensic analysis of cathinone drugs and metabolites

(For research use only, not for use in diagnostic procedures). In forensic drug analysis there is always a need for increased sensitivity to provide better detection of new synthetic substances in challenging casework. The capability to resolve very complex mixtures and provide confirmation of the identity of polar drugs and metabolites is the strength of Capillary Electrophoresis integrated with Mass Spectrometry. In this work we apply a sheathless interface for Capillary Electrophoresis-ESI-Mass Spectrometry (CESI-MS) for screening of 25 cathinone related designer drugs and metabolites in biofluids. The sample preparation included a pre-treatment step using only a simple dilution of urine. The CE used a 90 cm barefused silica, 150 μm o.d., 30 μm i.d. capillary, at 200-300 V/cm, 2.4 to 2.8 μamps at 25 °C. The background electrolyte (BGE) and conductive liquid were both 50 mM Ammonium Formate, pH 2.85. Sample introduction was either Hydrodynamic; 16s at 34.5 mBar or Electrokinetic; 10kV for 16s. The CE Instrument was a Beckman Coulter CESI 8000 coupled to an AB SCIEX 5600 TripleTOF® mass spectrometer through a sheathless interface High Sensitivity Porous Sprayer (HSPS). The ESI Voltage applied to the interface was 1.2 to 1.8 kV. The capillaries were initially conditioned with MeOH, water, 1N NaOH, water and BGE. A sheathless HSPS was used to interface CE and ESI-MS. The drugs tested included 25 cathinone drugs and metabolites in urine. Acquired MS/MS spectra were searched against an accurate mass drug library for confident compound identification. Combining high resolution MS and MS/MS acquisition with Capillary Electrophoresis separation, allows for retrospective compound identification of the cathinones with highest confidence based on accurate mass product ion specificity, mass spectral library matching as well as mass error, isotopic profile and molecular formula finding. The LOQ values for all components were found to be less than 5 ng/mL. Linearity was achieved between 5-1000 ng/mL for most drugs and % RSDs for different analytes are reported to be less than 10 % in most cases. A sheathless interface for Capillary Electrophoresis- ESI-Mass Spectrometry (CESI-MS) has shown to be a viable alternative for screening of 25 cathinone related designer drugs and metabolites in biofluids.

An LC–MS–MS method for quantitation of four new phenethylamines (BOX series) in plasma: in vivo application

The appearance of new “designer drugs” in the illicit market poses a serious health risk because they have unknown safety profiles, have a high potential for abuse, high potency, and can lead to devastating health consequences. For this reason, it is desirable to develop validated and reliable analytical screening tests that allow detection of amphetamines and related designer drugs in biological samples. We report a method for separation and quantitation of four new phenethylamines, 4-bromo-2,5-beta-trimethoxyphenethylamine (BOB), 4-methyl-2,5-beta-trimethoxyphenethylamine (BOD), 3,4-methylenedioxy-beta-methoxyphenethylamine (BOH), and 4-methyl-2,5-dimethoxy-beta-hydroxyphenethylamine (BOHD), in plasma. Quantitation was achieved via liquid chromatography–tandem mass spectrometry (LC–MS–MS) in the multiple reaction monitoring mode, using 2,3-dimethoxyphenethylamine-d3 as internal standard. The method was validated according to international guidelines. The parameters determined were selectivity, sensitivity, matrix effect, linearity, precision, recovery, and stability. All parameters were satisfactory. To remove matrix interference, solid-phase extraction was introduced in the method as clean-up step. The same method was applied in a pharmacokinetic study to monitor the target compounds in rat plasma after a single oral administration. The developed and validated LC–MS–MS method is the first available for quantitation of BOB, BOH, BOD, and BOHD in a biological matrix. This method is recommended for use in forensic and clinical toxicology, because of its sensitivity, selectivity, and simplicity. An important extension of this method could involve its application to other complex matrices.

Pharmacology and abuse of cocaine, amphetamines, Ecstasy and related designer drugs: a comprehensive review on their mode of action, treatment of abuse and intoxication

Pharmacology and abuse of cocaine, amphetamines, Ecstasy and related designer drugs: a comprehensive review on their mode of action, treatment of abuse and intoxication

A rapid method for the extraction, enantiomeric separation and quantification of amphetamines in hair

This paper presents a rapid and sensitive method for the determination and chiral separation of amphetamines and related designer drugs in hair samples. The substances are extracted from hair matrix by a 30 min treatment with a saturated carbonate buffer at pH 10 under ultrasonication. A commercial chiral derivatizing agent, trifluoroacetyl-prolyl chloride, is then added to the solution that is directly extracted with hexane and subsequently analyzed by GC/MS in SIM mode. R and S isomers of amphetamine, methamphetamine, MDA, MDMA and MDEA can be separated and detected with a limit of detection of 0.1 ng/mg for amphetamine, methamphetamine and MDA, and of 0.2 ng/mg for MDMA and MDEA. The method was then applied to 12 samples from suspected amphetamines abusers, showing the presence of both isomers of amphetamine and MDMA in one sample (27 and 1.5 ng/mg, respectively) and of MDMA in further eight samples, in concentrations ranging from traces to 2.7 ng/mg. No differences were observed in the disposition of different isomers in hair.

N-Methyl-1-(4-methoxyphenyl)-2-aminopropane (PMMA) and N-Methyl-1-(3,4-methylenedioxyphenyl)-2-aminopropane (MDMA) produce non-identical discriminative stimuli in rats

N-Methyl-1-(3,4-methylenedioxyphenyl)-2-aminopropane (methylenedioxymethamphetamine, MDMA, Ecstasy) and its structurally abbreviated congener N-methyl-1-(4-methoxyphenyl)-2-aminopropane ( para-methoxymethamphetamine, PMMA) are chemically related designer drugs, and PMMA is sometimes sold on the clandestine market as a substitute for MDMA. Prior drug discrimination studies have found that MDMA and PMMA substitute for one another suggesting that they produce similar discriminative stimulus effects in rats. However, there also are some indications that the two agents produce distinct stimulus effects. In this study, further comparisons were made between the stimulus effects of these two agents. Sprague–Dawley rats were trained to discriminate either 1.25 mg/kg of PMMA or 1.5 mg/kg of MDMA from saline vehicle in a two-lever operant paradigm. A structure–activity comparison revealed that MDMA and PMMA behave similarly upon homologation of their terminal amine substituents. In contrast, the PMMA stimulus, unlike an MDMA stimulus, failed to generalize completely to the psychostimulant cocaine, 8-hydroxy-2-( N, N-di- n-propylamino)tetralin (8-OH DPAT), and R(−)-1-(3-methoxyphenyl)-2-aminopropane [ R(−)MMA]. In an additional group of animals, a (+)amphetamine stimulus partially generalized to R(−)MMA. Taken together, the results argue and re-emphasize the conclusion that the stimulus effects produced by MDMA and PMMA are similar, but non-identical, and that PMMA is the less “stimulant-like” of the two.

Validation of the Cozart ® Amphetamine Microplate EIA for the analysis of amphetamines in oral fluid

The purpose of this study was to determine the performance characteristics of the Cozart ® Amphetamine Microplate EIA for detecting amphetamine in oral fluid. Oral fluid samples were collected using the Cozart ® RapiScan Collection System from 135 volunteer donors from drug treatment clinics. A further 35 oral fluid samples were collected from volunteer donors who were not drug users. The samples were analyzed in the laboratory using the Cozart ® Amphetamine Microplate EIA and confirmed using gas chromatography–mass spectrometry (GC–MS). The samples were stored frozen until analysis by GC–MS. The intra-assay precision for the Cozart ® Amphetamine Microplate EIA for amphetamine in oral fluid over forty assays was 2.74–7.1% CV (within assay) and 3.4–7.0% CV (within day). A total of 78 samples were positive for various amphetamines and related designer drugs. The Cozart ® Amphetamine Microplate EIA, using a cutoff of 45 ng/ml amphetamine equivalents in neat oral fluid, had a sensitivity of 91.7 ± 3.3% and a specificity of 95.9 ± 1.9% versus GC–MS using a cutoff of 30 ng/ml. A series of potential adulterants of oral fluid were evaluated and shown not to alter the outcome of the test result.

A rapid and sensitive liquid chromatography-tandem mass spectrometry method for the determination of amphetamine and related designer drugs in urine

Une methode pour l'analyse directe de six amphetaminiques dans les urines a ete developpee en utilisant la chromatographie liquide couplee a la spectrometrie de masse en tandem (LC-MS/MS). Nous avons ajoute 90 μl d'un melange d'etalons internes (1 pg/mL de d 5 -AMP, de d 5 -MET, de d 5 -MDA, de d 5 -MDMA, de d 5 -MDEA et de d 5 -MBDB) a 10 μl d'urine, melange par vortex et centrifuge. Les echantillons ont ete analyses par LC-MS/MS en mode MRM apres separation sur une colonne C18 a phase inverse en utilisant un gradient d'elution. La separation et la detection de tous les composes ont ete accomplies en huit minutes. La linearite a ete etablie pour tous les composes, de 78 a 100000 ng/mL. Les coefficients de correlation etaient superieurs a 0.998. Les limites de quantification etaient inferieures a 10 ng/mL, sauf pour l'amphetamine et la MDA (78 ng/mL). La repetabilite (CV%) et la reproductibilite variaient respectivement entre 2.62 et 16.26% et entre 0.86 et 11.98%. En ce qui concerne l'exactitude, le pourcentage de biais a 78 et 10000 ng/mL variait entre 0.16 et 7.17 %. La surface des pics des amphetamines ajoutees a de l'urine variait entre 85 et 100% de celle des amphetamines dissous dans un melange d'eau et de methanol, excepte pour l'amphetamine et le MDA. Le carry-over etait negligeable et la stabilite (156 et 5000 ng/mL) apres stockage a la temperature ambiante pendant 24h etait acceptable. En conclusion, la methode presentee permet la determination exacte, precise et rapide de six amphetaminiques dans les urines sur une plage de concentration large.

Advances in analytical toxicology: the current role of liquid chromatography-mass spectrometry in drug quantification in blood and oral fluid.

This paper reviews procedures for quantification of drugs in the biosamples blood, plasma, serum, or oral fluid (saliva, etc.) using liquid chromatography coupled with single-stage or tandem mass spectrometry (LC-MS, LC-MS-MS). Such procedures are important prerequisites for competent toxicological judgment and consultation in clinical and forensic toxicology. They cover blood (plasma, serum) analysis of amphetamines and related designer drugs, anesthetics, anticonvulsants, benzodiazepines, opioids, serotonergic drugs, tricyclic antidepressants, neuroleptics, antihistamines, beta-blockers, muscle relaxants, and sulfonylurea-type antidiabetics, and oral fluid analysis of amphetamines and related designer drugs, cocaine, benzoylecgonine, codeine, morphine, enantiomers of methadone and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), the nicotine metabolites cotinine and hydroxycotinine, and finally risperidone and its metabolite 9-hydroxyrisperidone. Basic information on the procedures is given in two tables and an example of quantification is illustrated in two figures. The pros and cons of such LC-MS procedures including sample work-up and ion suppression effects are critically discussed.

Identification of the human cytochromes P450 involved in the oxidative metabolism of “Ecstasy”-related designer drugs

The human cytochrome P450 (CYP) isozymes catalyzing the oxidative metabolism of the widely abused amphetamine derivatives MDMA ( N-methyl-3,4-methylenedioxyamphetamine, “Ecstasy”), MDE ( N-ethyl-3,4-methylenedioxyamphetamine, “Eve”), and MDA (3,4-methylenedioxyamphetamine) were identified. Using a simplified non-extractive reversed-phase HPLC assay with fluorescence detection, biphasic Michaelis–Menten kinetics were obtained for formation of all three dihydroxyamphetamines in liver microsomes from a CYP2D6 extensive metabolizer subject. In contrast, no low K m component was detectable in microsomes from a poor metabolizer subject. Additional specific probes for CYP2D6 further confirmed this isozyme as the exclusive low K m component for demethylenation. P450-selective inhibitors applied to CYP2D6-inhibited microsomes and activity measurements in a series of recombinant P450s suggested CYP1A2 as the major high K m component with contributions by CYP2B6 and CYP3A4. Moreover, the relative CYP1A2 content of a panel of 12 human livers was weakly but significantly correlated to the high K m demethylenase activity (Spearman rank correlation coefficient [r s] = 0.58; P < 0.05). Microsomal maximal velocities for N-dealkylation were at least 7-fold lower than for demethylenation and were characterized by apparently monophasic kinetics. The most important isozyme for this reaction appeared to be CYP2B6, the microsomal content of which was found to be strongly correlated to N-deethylation of MDE (r s = 0.90; P < 0.001). We conclude that, in addition to CP2D6 as the sole high-affinity demethylenase, several other P450 isozymes have the capacity to contribute to microsomal oxidative metabolism of methylenedioxyamphetamines. This may be of particular importance in individuals genetically lacking functional CYP2D6.