Methamphetamine And 3,4-methylenedioxymethamphetamine Research Articles

N, N-Dimethylpentylone (Dipentylone)-A New Synthetic Cathinone Identified in a Postmortem Forensic Toxicology Case Series.

Synthetic cathinones emerged on the novel psychoactive substance (NPS) drug market as alternatives to controlled stimulants and entactogens such as methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA). The majority of synthetic cathinones can be subclassified into two groups: beta-keto amphetamines (i.e., NPS with the suffix "drone") and beta-keto methylenedioxyamphetamines (i.e., NPS with the suffix "lone"). Although a significant number of beta-keto amphetamines have been identified, beta-keto methylenedioxyamphetamines have dominated the NPS market, including notable drugs like methylone, butylone, N-ethyl pentylone (ephylone), eutylone, and now N, N-dimethylpentylone. N, N-Dimethylpentylone, also known as dipentylone or bk-DMBDP, emerged into the illicit drug supply within two months of the international control of eutylone (September 2021). A novel standard addition method was developed and validated for N, N-dimethylpentylone, pentylone, and eutylone, and 18 postmortem cases were quantitated using the method described in this manuscript. The resulting blood concentration range for N, N-dimethylpentylone in this case series was 3.3 to 970 ng/mL (median: 145 ng/mL, mean: 277 ± 283 ng/mL). Pentylone, a metabolite of N, N-dimethylpentylone, was detected in all cases (range: 1.3-420 ng/mL, median: 31 ng/mL, and mean: 88 ± 127 ng/mL). Due to the rise in identifications of N, N-dimethylpentylone in postmortem investigations as well as the potential misidentification of N, N-dimethylpentylone as N-ethyl pentylone, samples testing positive for pentylone should be additionally confirmed for the presence of N, N-dimethylpentylone. Based on prior trends of new synthetic cathinones, it can be theorized that N, N-dimethylpentylone may predominate the U.S. synthetic stimulant market for the next one to two years, however; given the emergence of additional closely related isomeric compounds, it is important to utilize methodology capable of differentiating N, N-dimethylpentylone from its isomers (N-isopropylbutylone, N-ethyl pentylone, N-ethyl N-methyl butylone, hexylone, N-propylbutylone, diethylone, and tertylone).

Evaluation of Cytotoxic, Necrotic, Apoptotic, and Autophagic Effects of Methamphetamine and 3,4-Methylenedioxymethamphetamine on U-87 MG (Glial) and B104-1-1 (Neuronal) Cell Lines.

Methamphetamine (MA) and 3,4-methylenedioxymethamphetamine (MDMA) are empathogen (entactogen) psychoactive designer drugs which are mainly used for recreational purposes. Both MA and MDMA are central nervous system stimulants which are classified as monoamine neurotransmitter reuptake inhibitors. They have strong cytotoxic effects on dopaminergic and serotonergic neurons. Neurotoxicities of MA and MDMA by glial activation have been shown. The present work has investigated and measured cytotoxic, necrotic and apoptotic, and autophagic effects of MA and MDMA on U-87 MG (glial) and B104-1-1 (neuronal) cell lines by janus green, ethidium bromide/acridine orange, and monodansylcadaverine/propidium iodide staining to evaluate and compare their effects on glial and neuronal cells, respectively. The results of the present work showed that: (1) MDMA induced more potent mitochondrial toxicity, stronger necrotic and autophagic effects than MA in both B104-1-1 (neuronal) and U-87 MG (glial) cell lines; (2) although MDMA induced stronger apoptotic effect than MA on U-87 MG cell line, it had equal apoptotic effect on B104-1-1 cell line with MA; and (3) MDMA induced more potent mitochondrial toxicity, stronger necrotic, apoptotic, and autophagic effects than MA in B104-1-1 cell line than U-87 MG cell line.

Temporal monitoring of stimulants during the COVID-19 pandemic in Belgium through the analysis of influent wastewater.

Background and aimsWastewater-based epidemiology (WBE) is a complementary epidemiological data source to monitor stimulant consumption. The aims were to: (i) study intra- and inter-year temporal changes in stimulant use in Belgium during the first wave of the COVID-19 pandemic; and (ii) evaluate the effect of COVID-19 restrictive measures on stimulant consumption.MethodsThe study population corresponded to the catchments of four wastewater treatment plants corresponding with four Belgian cities (i.e., Antwerp-Zuid, Boom, Brussels, Leuven). Daily 24-h composite influent wastewater samples collected over one week in September 2019 and March through June 2020 during the first wave of the COVID-19 pandemic were analyzed for biomarkers of amphetamine, cocaine, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA). Measured concentrations were converted to population-normalized mass loads by considering the daily flow rate and the catchment population size. Mobile network data was used to accurately capture population movements in the different catchment areas. Temporal changes were assessed with multiple linear regression models, and the effect of the COVID-19 interventions on stimulant consumption were investigated.ResultsAn increase in amphetamine use was observed in three cities during governmental restrictions, with highest consumption predominantly during lockdown. Similarly, cocaine consumption was higher after the pandemic started, with highest consumption noted during the lockdown period in Boom and Leuven. Consumption of MDMA was similar in Antwerp-Zuid, Brussels and Leuven throughout the entire sampled period. In Boom, the highest consumption was observed during the full lockdown period.ConclusionsThe present study shows the potential of WBE to assess the impact of stringent lockdown measures on stimulant use in Belgium. This paper shows that strong restrictive measures did not have a profound effect on stimulant consumption.

Selective filtering of excitatory inputs to nucleus accumbens by dopamine and serotonin

The detailed mechanisms by which dopamine (DA) and serotonin (5-HT) act in the nucleus accumbens (NAc) to influence motivated behaviors in distinct ways remain largely unknown. Here, we examined whether DA and 5-HT selectively modulate excitatory synaptic transmission in NAc medium spiny neurons in an input-specific manner. DA reduced excitatory postsynaptic currents (EPSCs) generated by paraventricular thalamus (PVT) inputs but not by ventral hippocampus (vHip), basolateral amygdala (BLA), or medial prefrontal cortex (mPFC) inputs. In contrast, 5-HT reduced EPSCs generated by inputs from all areas except the mPFC. Release of endogenous DA and 5-HT by methamphetamine (METH) and (±)3,4-methylenedioxymethamphetamine (MDMA), respectively, recapitulated these input-specific synaptic effects. Optogenetic inhibition of PVT inputs enhanced cocaine-conditioned place preference, whereas mPFC input inhibition reduced the enhancement of sociability elicited by MDMA. These findings suggest that the distinct, input-specific filtering of excitatory inputs in the NAc by DA and 5-HT contribute to their discrete behavioral effects.

High concentrations of illicit stimulants and cutting agents cause false positives on fentanyl test strips

BackgroundThe opioid epidemic has caused an increase in overdose deaths which can be attributed to fentanyl combined with various illicit substances. Drug checking programs have been started by many harm reduction groups to provide tools for users to determine the composition of their street drugs. Immunoassay fentanyl test strips (FTS) allow users to test drugs for fentanyl by either filling a baggie or cooker with water to dissolve the sample and test. The antibody used in FTS is very selective for fentanyl at high dilutions, a characteristic of the traditional use of urine testing. These street sample preparation methods can lead to mg/mL concentrations of several potential interferents. We tested whether these concentrated samples could cause false positive results on a FTS.Methods20 ng/mL Rapid Response FTS were obtained from BTNX Inc. and tested against 4 different pharmaceuticals (diphenhydramine, alprazolam, gabapentin, and naloxone buprenorphine) and 3 illicit stimulants [cocaine HCl, methamphetamine, and 3,4-methylenedioxymethamphetamine (MDMA)] in concentrations from 20 to 0.2 mg/mL. The FTS testing pad is divided into 2 sections: the control area and the test area. Control and test area signal intensities were quantified by ImageJ from photographs of the test strips and compared to a threshold set by fentanyl at the FTS limit of detection.ResultsFalse positive results indicating the presence of fentanyl were obtained from samples of methamphetamine, MDMA, and diphenhydramine at concentrations at or above 1 mg/mL. Diphenhydramine is a common cutting agent in heroin. The street sample preparation protocols for FTS use suggested by many online resources would produce such concentrations of these materials. Street samples need to be diluted more significantly to avoid interference from potential cutting agents and stimulants.ConclusionsFentanyl test strips are commercially available, successful at detecting fentanyl to the specified limit of detection and can be a valuable tool for harm reduction efforts. Users should be aware that when drugs and adulterants are in high concentrations, FTS can give a false positive result.

Synthetic psychoactive cathinones: hypothermia and reduced lethality compared to methamphetamine and methylenedioxymethamphetamine

RationaleSynthetic psychoactive cathinones (SPCs) are drugs with psychostimulant and entactogenic properties like methamphetamine (MA) and 3,4-methylenedioxymethamphetamine (MDMA). Despite clinical reports of human overdose, it remains to be determined if SPCs have greater propensity for adverse effects than MA or MDMA. ObjectivesTo determine whether the SPCs cathinone (CAT), methcathinone (MCAT), mephedrone (MMC), and methylenedioxypyrovalerone (MDPV) have lower LD50 values than MA or MDMA. MethodsMale and female C57Bl/6J mice received single injections of one of 6 doses of a test drug (0–160 mg/kg IP). Temperature and behavioral observations were taken every 20 min for 2 h followed by euthanasia of surviving mice. Organs were weighed and evaluated for histopathological changes. ResultsLD50 values for MA and MDMA, 84.5 and 100.9 mg/kg respectively, were similar to previous observations. The LD50 for MMC was 118.8 mg/kg, but limited lethality was observed for other SPCs (CAT, MCAT, MDPV), so LD50 values could not be calculated. For all drugs, death was associated with seizure, when it was observed. Rather than hyperthermia, dose-dependent hypothermia was observed for MMC, MDPV, CAT, and MCAT. Contrary to initial expectations, none of the SPCs studied here had LD50 values lower than MA or MDMA. ConclusionsThese data indicate that, under the conditions studied here: (1) SPCs exhibit less lethality than MA and MDMA; (2) SPCs impair thermoregulation; (3) effects of SPCs on temperature appear to be independent of effects on lethality.

A simplified fabric phase sorptive extraction method for the determination of amphetamine drugs in water samples using liquid chromatography-mass spectrometry.

Fabric phase sorptive extraction (FPSE) can directly extract the target analytes and simultaneously determine many similar substances from complicated sample matrices. Also, it has very high chemical stability. Therefore, we used fabric phase sorptive extraction to analyze three amphetamine drugs (amphetamine (AM), methamphetamine (MAM), and 3,4-methylenedioxymethamphetamine (MDMA)) in water. This was coupled with ultrahigh-performance liquid chromatography and tandem mass spectrometry. The effects of different sorbent chemistries such as sorption time, ratios of back-extraction solvents, back-extraction time, and the salt effect on the extraction efficiency were studied; the optimum operation conditions were determined. Medium polarity polar polymer-coated FPSE media were created using short-chain poly (tetrahydrofuran) (PTHF). This is the most efficient extraction media for the analytes of interest. Under the optimized conditions, the linear range of the three amphetamine drugs were 0.1–150.0 (AM, MAM) and 0.5–200 ng mL−1 (MDMA). The correlation coefficients (γ) were 0.9947 (AM), 0.9925 (MAM), and 0.9918 (MDMA). The detection limits (LOD) were 0.025 ng mL−1 for AM, 0.029 ng mL−1 for MAM, and 0.01 ng mL−1 for MDMA. The corresponding limit of quantification values (LOQ) were 0.083 ng mL−1, 0.097 ng mL−1, and 0.031 ng mL−1, respectively. The recoveries were 73.4–91.6%, 82.6–95.4%, and 92.7–95.3%, respectively, and the relative standard deviations (RSD) were 1.65–6.88%, 1.38–6.11%, and 1.58–7.34%, respectively. Moreover, our method can be successfully applied for the analysis of amphetamines in wastewater samples, and at the same time, lays the foundation for the future detection of such substances.

Spatio-temporal assessment of illicit drug use at large scale: evidence from 7 years of international wastewater monitoring.

Background and aimsWastewater‐based epidemiology is an additional indicator of drug use that is gaining reliability to complement the current established panel of indicators. The aims of this study were to: (i) assess spatial and temporal trends of population‐normalized mass loads of benzoylecgonine, amphetamine, methamphetamine and 3,4‐methylenedioxymethamphetamine (MDMA) in raw wastewater over 7 years (2011–17); (ii) address overall drug use by estimating the average number of combined doses consumed per day in each city; and (iii) compare these with existing prevalence and seizure data.DesignAnalysis of daily raw wastewater composite samples collected over 1 week per year from 2011 to 2017.Setting and ParticipantsCatchment areas of 143 wastewater treatment plants in 120 cities in 37 countries.MeasurementsParent substances (amphetamine, methamphetamine and MDMA) and the metabolites of cocaine (benzoylecgonine) and of Δ9‐tetrahydrocannabinol (11‐nor‐9‐carboxy‐Δ9‐tetrahydrocannabinol) were measured in wastewater using liquid chromatography–tandem mass spectrometry. Daily mass loads (mg/day) were normalized to catchment population (mg/1000 people/day) and converted to the number of combined doses consumed per day. Spatial differences were assessed world‐wide, and temporal trends were discerned at European level by comparing 2011–13 drug loads versus 2014–17 loads.FindingsBenzoylecgonine was the stimulant metabolite detected at higher loads in southern and western Europe, and amphetamine, MDMA and methamphetamine in East and North–Central Europe. In other continents, methamphetamine showed the highest levels in the United States and Australia and benzoylecgonine in South America. During the reporting period, benzoylecgonine loads increased in general across Europe, amphetamine and methamphetamine levels fluctuated and MDMA underwent an intermittent upsurge.ConclusionsThe analysis of wastewater to quantify drug loads provides near real‐time drug use estimates that globally correspond to prevalence and seizure data.

Understanding the Removal and Fate of Selected Drugs of Abuse in Sludge and Biosolids from Australian Wastewater Treatment Operations

Abstract Illicit and pharmaceutical drugs are considered to be emerging contaminants of concern, and much research effort has gone into assessing their occurrence in wastewater. However, little information exists on their presence in treated sludge or biosolids. In this study, we examined sludge and biosolids from a large metropolitan wastewater treatment plant (WWTP) in Australia to determine the occurrence of five drugs of abuse, including benzoylecgonine as indicator of cocaine consumption, methamphetamine and 3,4-methylenedioxy methamphetamine (MDMA) as representative illicit stimulants, and codeine and morphine as pharmaceuticals with potential environmental risk. The samples were solid-phase extracted and analyzed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Benzoylecgonine and MDMA were present in raw sludge but were notably degraded during solids treatment processes, and were not detected in the dewatered sludge (after treatment) or in biosolids. Methamphetamine, codeine, and morphine were detected in all biosolids samples at mean concentrations of 20–50 μg·kg−1. The presence of these three drugs in biosolids shows that these compounds are relatively stable in the solids and in soil, and can persist in biosolids for at least several years. A simple environmental risk assessment based on estimated risk quotients (RQs) for these compounds indicated that the potential environmental risks associated with the land application of biosolids are very low at typical Australian biosolids application rates.

Do cosmetic treatments increase the incorporation of amphetamines into hair?

According to several studies, the concentration of drugs in hair may decrease by bleaching or dyeing; however, at the same time, treated hair could incorporate greater quantity of drugs from sweat or sebum than intact hair. The aim of this study is to assess the incorporation of amphetamine (AP), methamphetamine (MA) and 3,4-methylenedioxy-methamphetamine (MDMA) from sweat or sebum to virgin, dyed, and bleached hair. Drug-free head hair samples (from 20 volunteers) were divided into four locks, one of them was left untreated, another was subjected to dyeing treatment, and the last two were subjected to different degree of bleaching. Virgin, dyed, mildly as well as severely bleached locks of hair were exposed to artificial sweat or sebum containing AP, MA, and MDMA (500 ng/g), at 37 °C for a time period of 8 and 24 h, respectively. After the exposure, all the hair samples were tested by GC/MS in SIM mode, applying an already published method. Drug concentrations in hair ranged 0.41 to 2.20 ng/mg from artificial sweat, while only reached 0.27 ng/mg from artificial sebum. The differences of each drug concentration (AP, MA, MDMA) between different kind of hair were statistically compared: virgin hair vs. dyed hair, virgin hair vs mildly bleached hair, virgin hair vs. severely bleached hair. The differences between different drugs concentration within each kind of hair (virgin, dyed, mildly and severely bleached) were, as well, statistically compared: AP vs. MA, AP vs. MDMA, MA vs. MDMA. In all cases, differences were calculated between locks of hair from the same volunteer. The paired samples Student's t-test was applied in cases where the concentration differences followed the normal law. In these cases the extent of the difference was calculated with a confidence interval of 95%. The paired samples Wilcoxon's t -test was applied in cases where the concentration differences did not follow the normal law. Regarding the incorporation of AP, MA and MDMA from artificial sweat in vitro, the analysis provides evidence that the mean concentration for each drug in dyed or bleached hair is higher than the mean concentration in virgin hair, and that the differences between means are likely to be between 0.15 and 0.57 ng/mg, depending on the drug and the compared pairs. The incorporation of different drugs in the same kind of hair shows little differences in virgin and dyed hair, but the mean concentration of MDMA in bleached hair is higher than the mean concentration of AP; the differences between means are likely to be between 0.08 and 0.16 ng/mg. Regarding the incorporation of AP, MA and MDMA from artificial sebum in vitro, the differences between mean concentrations are lower for every compared pair, in relation to the incorporation from sweat. AP is not incorporated into hair from sweat or sebum to a greater extent than MDMA. Higher positivity rates for AP have been found in dyed or bleached hair in relation to virgin hair, detecting both significant [1] and non-significant [2] differences in the target populations. Based on these facts and the quantitative results of in vitro incorporation of amphetamine derivatives in hair, when interpreting the results from cosmetically treated hair, consideration should be given not only to the possibility that the treatment will lead to a decrease in drug concentration, but also to an increase. This consideration is particularly relevant if the detected levels are close to the cut-off.

Rapid and simple analysis of amphetamine-type illegal drugs using excitation–emission matrix fluorescence coupled with parallel factor analysis

ABSTRACTNowadays, the abuse of illegal drugs has been an increasingly grim problem in the world. Excitation–emission matrix fluorescence combined with parallel factor analysis was used to make a quantitative analysis of the simulated amphetamine-type illegal drugs. Satisfactory results were achieved for simultaneous determination of methamphetamine (MAM) and 3, 4-methylenedioxymethamphetamine (MDMA) in the presence of adulterants. The average recoveries were (99.8 ± 0.6)% and (101.6 ± 5.7)% for MAM and MDMA, respectively. Figures of merit including root-mean-square error of calibration and prediction, sensitivity and selectivity were investigated to evaluate the performance of the proposed method. The limits of detection were 0.054 and 0.002 1 μg/mL for MAM and MDMA, respectively.

Investigation of the effects of ‘piperazine-containing party pills’ and dexamphetamine on interhemispheric communication using electroencephalography

'Piperazine-containing party pills' were marketed and sold as legal alternatives to methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) until 2008 in New Zealand. The major constituents of these 'pills' were benzylphenylpiperazine (BZP) and trifluoromethylphenylpiperazine (TFMPP). Despite their popularity, there is a paucity of knowledge about their central effects in humans. This study investigated their effects on human neural processing using electroencephalographic techniques. A randomised, double-blind, placebo-controlled study investigated the effects of an acute dose of these compounds on the interhemispheric transfer of information (IHTT) using the Poffenberger task. Reaction time data were also collected. Healthy, right-handed males were given an oral dose of either BZP (n = 13) (200mg), TFMPP (n = 15) (60mg), a combination of BZP + TFMPP (n = 15) (100mg/30mg), dexamphetamine (n = 16) (20mg), or placebo (n = 23) and tested both before and 120min after drug administration. A mixed factorial repeated measures analysis of variance of absolute N160 latency and contrast analysis revealed that only TFMPP (F (1,77) = 17.30, p ≤ 0.001) significantly reduced the absolute N160 latency. Analysis of the IHTT revealed that only TFMPP (F (1,77) = 5.266, p ≤ 0.02) significantly reduced the IHTT, while BZP, BZP + TFMPP and dexamphetamine had no effect. Contrast analysis revealed that both TFMPP (F (1,77) = 17.30, p ≤ 0.001) and placebo (F (1,77) = 15.08, p ≤ 0.001) preserved the laterality of information transfer from one hemisphere to the other. Reaction time (p > 0.05) was not significantly affected by any of the drug treatments. The usual directional asymmetry (i.e. faster R-to-L transfer relative to L-to-R) observed in healthy control group was absent following the administration of either BZP, BZP + TFMPP or dexamphetamine. Surprisingly, lateralised hemispheric function was not affected by TFMPP. Our findings highlight how the administration of BZP, TFMPP and BZP + TFMPP leads to changes in the pattern of information transfer.

Three-dimensional surface-enhanced Raman scattering hotspots in spherical colloidal superstructure for identification and detection of drugs in human urine.

Rapid component separation and robust surface-enhanced Raman scattering (SERS) identification of drugs in real human urine remain an attractive challenge because of the sample complexity, low molecular affinity for metal surface, and inefficient use of hotspots in one- or two-dimensional (2D) geometries. Here, we developed a 5 min strategy of cyclohexane (CYH) extraction for separating amphetamines from human urine. Simultaneously, an oil-in-water emulsion method is used to assemble monodisperse Ag nanoparticles in the CYH phase into spherical colloidal superstructures in the aqueous phase. These superstructures create three-dimensional (3D) SERS hotspots which exist between every two adjacent particles in 3D space, break the traditional 2D limitation, and extend the hotspots into the third dimension along the z-axis. In this platform, a conservative estimate of Raman enhancement factor is larger than 10(7), and the same CYH extraction processing results in a high acceptability and enrichment of drug molecules in 3D hotspots which demonstrates excellent stability and reproducibility and is suitable for the quantitative examination of amphetamines in both aqueous and organic phases. Parallel ultraperformance liquid chromatography (UPLC) examinations corroborate an excellent performance of our SERS platform for the quantitative analysis of methamphetamine (MA) in both aqueous solution and real human urine, of which the detection limits reach 1 and 10 ppb, respectively, with tolerable signal-to-noise ratios. Moreover, SERS examinations on different proportions of MA and 3,4-methylenedioxymethamphetamine (MDMA) in human urine demonstrate an excellent capability of multiplex quantification of ultratrace analytes. By virtue of a spectral classification algorithm, we realize the rapid and accurate recognition of weak Raman signals of amphetamines at trace levels and also clearly distinguish various proportions of multiplex components. Our platform for detecting drugs promises to be a great prospect for a rapid, reliable, and on-spot analyzer.

Adsorption and desorption characteristics of methamphetamine, 3,4-methylenedioxymethamphetamine, and pseudoephedrine in soils.

This work presents, for the first time, information on the adsorption-desorption characteristics of illicit drugs and precursors in soils and an estimation of their potential bioavailability. The experiment was conducted using a batch equilibrium technique for the parent drugs methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) and the precursor pseudoephedrine in three South Australian soils varying in physiochemical properties. The individual compounds exhibited different adsorption mechanisms in the test soils, and the results fitted better with the Freundlich isotherm model (r (2) ≥ 0.99). The maximum adsorption capacity was recorded for pseudoephedrine (2,000 μg g(-1)). However, pseudoephedrine recorded lower organic carbon normalized adsorption coefficient values (<250 mL g(-1)), lower magnitudes of Gibb's free energy change, and higher percent desorption (73-92 %) compared to methamphetamine and MDMA. The results thus showed pseudoephedrine to be the most mobile compound in the soils under study, to have the highest availability for degradation of the three compounds, and to have the highest susceptibility to biotic degradation in test soils.

Effects of combined treatment with mephedrone and methamphetamine or 3,4-methylenedioxymethamphetamine on serotonin nerve endings of the hippocampus

AimsMephedrone is a stimulant drug of abuse with close structural and mechanistic similarities to methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA). Although mephedrone does not damage dopamine nerve endings it increases the neurotoxicity of amphetamine, methamphetamine and MDMA. The effects of mephedrone on serotonin (5HT) nerve endings are not fully understood, with some investigators reporting damage while others conclude it does not. Presently, we investigate if mephedrone given alone or with methamphetamine or MDMA damages 5HT nerve endings of the hippocampus. Main methodsThe status of 5HT nerve endings in the hippocampus of female C57BL mice was assessed through measures of 5HT by HPLC and by immunoblot analysis of serotonin transporter (SERT) and tryptophan hydroxylase 2 (TPH2), selective markers of 5HT nerve endings. Astrocytosis was assessed through measures of glial fibrillary acidic protein (GFAP) (immunoblotting) and microglial activation was determined by histochemical staining with Isolectin B4. Key findingsMephedrone alone did not cause persistent reductions in the levels of 5HT, SERT or TPH2. Methamphetamine and MDMA alone caused mild reductions in 5HT but did not change SERT and TPH2 levels. Combined treatment with mephedrone and methamphetamine or MDMA did not change the status of 5HT nerve endings to an extent that was different from either drug alone. SignificanceMephedrone does not cause toxicity to 5HT nerve endings of the hippocampus. When co-administered with methamphetamine or MDMA, drugs that are often co-abused with mephedrone by humans, toxicity is not increased as is the case for dopamine nerve endings when these drugs are taken together.

Using quantitative wastewater analysis to measure daily usage of conventional and emerging illicit drugs at an annual music festival

Wastewater analysis provides a non-intrusive way of measuring drug use within a population. We used this approach to determine daily use of conventional illicit drugs [cannabis, cocaine, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA)] and emerging illicit psychostimulants (benzylpiperazine, mephedrone and methylone) in two consecutive years (2010 and 2011) at an annual music festival. Daily composite wastewater samples, representative of the festival, were collected from the on-site wastewater treatment plant and analysed for drug metabolites. Data over 2 years were compared using Wilcoxon matched-pair test. Data from 2010 festival were compared with data collected at the same time from a nearby urban community using equivalent methods. Conventional illicit drugs were detected in all samples whereas emerging illicit psychostimulants were found only on specific days. The estimated per capita consumption of MDMA, cocaine and cannabis was similar between the two festival years. Statistically significant (P < 0.05; Z = -2.0-2.2) decreases were observed in use of methamphetamine and one emerging illicit psychostimulant (benzyl piperazine). Only consumption of MDMA was elevated at the festival compared with the nearby urban community. Rates of substance use at this festival remained relatively consistent over two monitoring years. Compared with the urban community, drug use among festival goers was only elevated for MDMA, confirming its popularity in music settings. Our study demonstrated that wastewater analysis can objectively capture changes in substance use at a music setting without raising major ethical issues. It would potentially allow effective assessments of drug prevention strategies in such settings in the future.

Psychoactive ‘Bath Salts’: Compounds, Mechanisms, and Toxicities

Recently, there has been an alarming increase in the abuse of so-called ‘bath salts' products sold on the internet and in retail shops. These products have no legitimate use as bath additives, rather they are purchased as legal alternatives to illicit drugs like cocaine, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) (Prosser and Nelson, 2012; Spiller et al, 2011). Bath salts powders are self-administered by insufflation (ie, snorted), taken orally or injected intravenously (i.v.). Recreational doses of bath salts enhance mood and increase alertness, whereas higher doses can lead to dangerous neurological and cardiovascular complications. The psychoactive constituents in bath salts have been identified as synthetic derivatives of cathinone, an amphetamine-type stimulant (Shanks et al, 2012; Spiller et al, 2011). Figure 1 illustrates the chemical structures of three popular bath salts compounds: 4-methylmethcathinone (mephedrone), 3,4-methylenedioxymethcathinone (methylone) and 3,4-methylenedioxypyrovalerone (MDPV). Owing to the public health risks posed by bath salts, the US government placed mephedrone, methylone and MDPV into Schedule I control in November 2011 (DEA, 2011). Unfortunately, a new wave of cathinone derivatives has appeared in the marketplace to replace those drugs now subject to regulatory control (Shanks et al, 2012). Figure 1 Chemical structures of bath salts cathinones and related compounds. Despite the widespread use of bath salts, there is a paucity of information about their pharmacology. Using assay methods in rat brain synaptosomes, it has been shown that mephedrone and methylone act as substrates at plasma membrane transporters for norepinephrine, dopamine, and serotonin, thereby stimulating non-exocytotic release of these neurotransmitters (Baumann et al, 2012). Mephedrone is about two-times more potent than methylone as a transporter substrate. In vivo microdialysis studies in rats confirm that mephedrone and methylone (0.3 or 1.0 mg/kg, i.v.) increase extracellular levels of dopamine and serotonin in the brain, similar to the effects of MDMA. Little is known about the pharmacology of MDPV, but our unpublished findings show the drug is a potent blocker of dopamine and norepinephrine uptake. The fact that synthetic cathinones enhance dopamine transmission predicts high abuse liability. Hadlock et al (2011) reported that i.v. mephedrone (0.24 mg/infusion) is readily self-administered by rats, but reinforcing effects of other synthetic cathinones are largely unexplored. Few preclinical studies have examined the pharmacokinetics and metabolism of synthetic cathinones or the consequences of chronic drug dosing, and these types of investigations are needed. Serotonin transporter substrates like MDMA are known to produce sustained deficits in brain serotonin neurons of laboratory animals, so mephedrone and methylone could have similar actions. Repeated subcutaneous (s.c.) administration of either drug to single-housed rats (3 or 10 mg/kg, three doses) has no long-lasting effects on brain tissue monoamines (Baumann et al, 2012), but administration of higher doses of mephedrone to group-housed rats (10 or 25 mg/kg, s.c., four doses) causes selective depletion of brain serotonin (Hadlock et al, 2011). It is noteworthy that MDPV is the chief substance detected in blood and urine from patients hospitalized for bath salts overdose in the US (Spiller et al, 2011). Such patients display agitation, combative behavior, hallucinations, delusions, hyperthermia, tachycardia and hypertension (Prosser and Nelson, 2012; Spiller et al, 2011). Health care workers should be aware that patients presenting with this constellation of symptoms may have taken bath salts, and appropriate supportive care should be provided. Owing to the increasing availability of ‘replacement' cathinones with unknown pharmacology (Shanks et al, 2012), it seems likely that emergency departments will continue to encounter patients suffering from adverse effects of synthetic cathinone abuse.

Evaluation of hair roots for detection of methamphetamine and 3,4-methylenedioxymethamphetamine abuse by use of an HPLC–chemiluminescence method

We describe the use of hair roots as a matrix for detection of methamphetamine (MP) and 3,4-methylenedioxymethamphetamine (MDMA) abuse. The concentration of drugs was determined in rat hair roots, hair shafts, and plasma after a single administration of MP or MDMA, by use of an HPLC-peroxyoxalate chemiluminescence (PO-CL) method involving column switching. Plasma and hair roots and shafts were collected from male Wistar rats before and after administration of MP (10 mg kg(-1), i.p.). In addition, the roots and shafts of pigmented and non-pigmented hair of male Lister hooded rats were collected after administration of MDMA (10 mg kg(-1), i.p.). The concentrations of MP and MDMA in plasma and hair were determined by use of the HPLC-PO-CL method, with satisfactory sensitivity and reproducibility. The concentration of MP in hair roots 1-14 days after administration ranged from 0.038 to 0.115 ng mg(-1) (n = 3). By use of the HPLC-PO-CL method, MP could be detected in hair roots for longer (up to 14 days) than it could be detected in conventional biological specimens, for example plasma (~1 day), and MDMA was detected in hair roots from 1 to 10 days after administration. The AUC(1-10) (ng day mg(-1)) for MDMA in roots of non-pigmented and pigmented hair was comparable (4.93 ± 2.09 vs. 6.67 ± 1.28, n = 3), whereas AUC(1-14) for hair shafts differed significantly (1.86 ± 0.93 vs. 4.58 ± 0.63, P < 0.05, n = 3). The window for detecting MP (or MDMA) in hair roots under our conditions was 1-14 (or 1-10) days.

Development of a simultaneous analytical method for selected anorectics, methamphetamine, MDMA, and their metabolites in hair using LC-MS/MS to prove anorectics abuse

Owing to the tight control of methamphetamine, it is presumed that phentermine, an amphetamine-type anorectic, has recently been considered a supplement for methamphetamine abusers in Korea. In addition, the abuse of other anorectics obtained by inappropriate means has become a social issue. Hair is a useful specimen to prove chronic drug use. Therefore, an analytical method for the simultaneous detection of phentermine, phendimetrazine, amfepramone, fenfluramine, mazindol, methamphetamine, and 3,4-methylenedioxymethamphetamine (MDMA), as well as their metabolites, which covers the major amphetamines and anorectic agents in Korea, in hair was established and validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The drugs and their metabolites in hair were extracted using 1 % HCl in methanol and then filtered and analyzed by LC-MS/MS with electrospray ionization in positive mode. The validation results for selectivity, linearity, matrix effect, recovery, process efficiency, intra- and interassay precision and accuracy, and processed sample stability were satisfactory. The limits of detection ranged from 0.025 to 1 ng/10 mg hair and the limits of quantification were 0.25 ng/10 mg hair for every analyte except mazindol and phentermine, for which they were 10 ng/10 mg hair. The method was successfully applied for the segmental determination of selected anorectics, methamphetamine, MDMA, and their metabolites in hair from 39 drug suspects. Among the anorectics, phentermine and/or phendimetrazine were identified with or without methamphetamine in the hair samples. Closer supervision of the inappropriate use of anorectics is necessary. Also, hair analysis is useful for monitoring the abuse potential of unnoticed drugs.

MDMA, methamphetamine, and CYP2D6 pharmacogenetics: what is clinically relevant?

In vitro human studies show that the metabolism of most amphetamine-like psychostimulants is regulated by the polymorphic cytochrome P450 isozyme CYP2D6. Two compounds, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA), were selected as archetypes to discuss the translation and clinical significance of in vitro to in vivo findings. Both compounds were chosen based on their differential interaction with CYP2D6 and their high abuse prevalence in society. Methamphetamine behaves as both a weak substrate and competitive inhibitor of CYP2D6, while MDMA acts as a high affinity substrate and potent mechanism-based inhibitor (MBI) of the enzyme. The MBI behavior of MDMA on CYP2D6 implies that subjects, irrespective of their genotype/phenotype, are phenocopied to the poor metabolizer (PM) phenotype. The fraction of metabolic clearance regulated by CYP2D6 for both drugs is substantially lower than expected from in vitro studies. Other isoenzymes of cytochrome P450 and a relevant contribution of renal excretion play a part in their clearance. These facts tune down the potential contribution of CYP2D6 polymorphism in the clinical outcomes of both substances. Globally, the clinical relevance of CYP2D6 polymorphism is lower than that predicted by in vitro studies.