Showing papers in "Drug Testing and Analysis in 2014"

From PCP to MXE: a comprehensive review of the non-medical use of dissociative drugs.

Abstract: PCP or phencyclidine was discovered in 1956 and soon became a popular street drug. Dissociatives including PCP, ketamine, and dextromethorphan have been used non-medically for their mind-altering effects for over 60 years. Many of these compounds have also been used clinically and in legitimate research. At least 14 derivatives of PCP were sold for non-medical and illict use from the late 1960s until the 1990s. With the advent of the Internet, the drug market underwent a dramatic evolution. While initially gray-market chemical vendors offering dextromethorphan and ketamine thrived, most recently the market has shifted to legal high and online-based research chemical vendors. Starting with the first dissociative research chemical, 4-MeO-PCP in 2008, the dissociative research chemical market has rapidly evolved and currently comprises at least 12 dissociatives, almost half of which were unknown in the scientific literature prior to their introduction. Several of these, including methoxetamine, have reached widespread use internationally. A historical account of non-medical use of over 30 dissociative compounds was compiled from a diverse collection of sources. The first complete portrait of this underground market is presented along with the relevant legal, technological, and scientific developments which have driven its evolution.

The adverse health effects of chronic cannabis use

Abstract: This paper summarizes the most probable of the adverse health effects of regular cannabis use sustained over years, as indicated by epidemiological studies that have established an association between cannabis use and adverse outcomes; ruled out reverse causation; and controlled for plausible alternative explanations. We have also focused on adverse outcomes for which there is good evidence of biological plausibility. The focus is on those adverse health effects of greatest potential public health significance – those that are most likely to occur and to affect a substantial proportion of regular cannabis users. These most probable adverse effects of regular use include a dependence syndrome, impaired respiratory function, cardiovascular disease, adverse effects on adolescent psychosocial development and mental health, and residual cognitive impairment. Copyright © 2013 John Wiley & Sons, Ltd.

A review of the cultivation and processing of cannabis (Cannabis sativa L.) for production of prescription medicines in the UK.

Abstract: The quality demands of the pharmaceutical industry require prescription medicines to be consistent in their active ingredient content. Achieving this, using raw cannabis as a feedstock, is especially challenging. The plant material is extremely inhomogeneous, and the ratios of active ingredients are affected by a range of factors. These include the genetics of the plant, the growing and storage conditions, the state of maturity at harvest, and the methods used to process and formulate the material. The reasons for this variability are described, with particular emphasis on the botanical considerations. To produce the complex botanical medicine Sativex®, which contains the cannabinoids Δ(9)-tetrahydrocannabinol (THC) and cannabidiol (CBD) and a range of other ingredients, GW Pharmaceuticals had to manage these variables. This medicine, for the treatment of spasticity due to multiple sclerosis, is the first cannabis-based medicine to be approved in the UK. The company's methodology for producing this and other chemotypes is described.

Dried blood spots: concepts, present status, and future perspectives in bioanalysis.

Abstract: Over the past several years, dried blood spot (DBS) sampling technique has emerged as a pertinent method in both qualitative and quantitative bioanalysis context. In the DBS method, the blood sample is directly soaked on to a paper (with or without treatment). After drying it can be analyzed by modern analytical, immunological, or genomic detection systems. Several advantages of the DBS technique such as low blood volume requirement, transportation and storage without special treatment, better analytes stability, enhanced clinical cooperation in clinical trials, and reduced unforeseeable exposure of analysts to biohazards, make it the most appropriate blood sampling technique. This review illustrates the information available on the DBS method which may serve as a single window for investigators in the field of bioanalysis. Also, it explores the proficiency and appliance of the DBS method in pharmacokinetic (PK), therapeutic drug monitoring (TDM), toxicokinetic (TK), metabolomic, and disease diagnosis.

New psychoactive substances as adulterants of controlled drugs. A worrying phenomenon

Abstract: The use of new psychoactive substances (NPS) as adulterants has received little attention in the literature. In this paper, results from Energy Control's drug checking service documenting the use of NPS as adulterants of controlled drugs are presented, and some reflections about possible explanations for this new phenomenon, potential risks for users, and challenges that it poses are discussed. From 2009 to 2012, 24 NPS belonging to several chemical classes such as phenethylamines, substituted cathinones, tryptamines, and methoxetamine were identified in 173 samples believed to be MDMA, amphetamine, ketamine, cocaine, mescaline, or methamphetamine. The NPS adulterant most frequently observed was 2-(4-bromo-2,5-dimethoxyphenyl)ethanamine (2C-B) followed by 1-(4-fluorophenyl)propan-2-amine (4-FA). Sixty-nine different combinations of substances were detected: 20 involving a controlled drug combined with an NPS, and 49 involving one or more NPS that substituted the controlled drug. As these combinations could pose substantial risks to users, the need to improve knowledge about toxicity associated with these combinations, and the danger of these substances being incorporated into the products of illegal markets, are highlighted. Drug checking services and the European Union's early-warning system operated by the European Monitoring Centre for Drugs and Drug Addiction (EMCDDA) and Europol can play an important role in reducing the harm associated with this phenomenon.

Changes in the prevalence of new psychoactive substances before and after the introduction of the generic scheduling of synthetic cannabinoids in Japan

Abstract: To counter the spread of the many analogues of psychoactive substances, the Pharmaceutical Affairs Law in Japan was amended in 2006 to establish a new category - Designated Substances - in order to more promptly control these drugs. As of March 2013, 106 substances (including one plant, Salvia divinorum) were listed in the category of Designated Substances, and 13 of them had had their category changed from Designated Substances into the much stricter category, Narcotics. However, new analogues of controlled substances, especially synthetic cannabinoids, appeared one-by-one since the new category was introduced. To avoid a cat-and-mouse game between regulators and illicit drug manufacturers, a comprehensive system (generic scheduling) for designating naphthoylindole-type synthetic cannabinoids, with particular substituents, was introduced into the Designated Substances in 2013. Since late 2012, the naphthoylindole-type compounds have been gradually replaced by other types of synthetic cannabinoids, such as cyclopropylmethanones, cannabimimetic carboxamide derivatives, adamanthoyl indoles, and cannabimimetic quinolinyl carboxylates. After the enforcement of the generic scheduling for designating naphthoylindoles in March 2013, these naphthoylindoles have been completely replaced by other types and have rarely been detected in the products. New types of psychoactive substances, including opioid receptor agonists (e.g. AH-7921, MT-45), hallucinogenic phenethylamines (e.g. NBOMe-type compounds), and thiophene derivatives (e.g. methiopropamine, α-PVT) have also appeared. The almost infinite possibilities of altered structures of chemicals make it difficult to carry out effective and exhaustive scheduling. To prevent the widespread distribution and abuse of these new psychoactive substances, continuous and dedicated monitoring for the emergence of these substances is necessary.

Therapeutic potential of cannabinoid medicines

Abstract: Cannabis was extensively used as a medicine throughout the developed world in the nineteenth century but went into decline early in the twentieth century ahead of its emergence as the most widely used illicit recreational drug later that century. Recent advances in cannabinoid pharmacology alongside the discovery of the endocannabinoid system (ECS) have re-ignited interest in cannabis-based medicines. The ECS has emerged as an important physiological system and plausible target for new medicines. Its receptors and endogenous ligands play a vital modulatory role in diverse functions including immune response, food intake, cognition, emotion, perception, behavioural reinforcement, motor co-ordination, body temperature, wake/sleep cycle, bone formation and resorption, and various aspects of hormonal control. In disease it may act as part of the physiological response or as a component of the underlying pathology. In the forefront of clinical research are the cannabinoids delta-9-tetrahydrocannabinol and cannabidiol, and their contrasting pharmacology will be briefly outlined. The therapeutic potential and possible risks of drugs that inhibit the ECS will also be considered. This paper will then go on to review clinical research exploring the potential of cannabinoid medicines in the following indications: symptomatic relief in multiple sclerosis, chronic neuropathic pain, intractable nausea and vomiting, loss of appetite and weight in the context of cancer or AIDS, psychosis, epilepsy, addiction, and metabolic disorders.

Current knowledge on cannabinoids in oral fluid

Abstract: Oral fluid (OF) is a new biological matrix for clinical and forensic drug testing, offering non-invasive and directly observable sample collection reducing adulteration potential, ease of multiple sample collections, lower biohazard risk during collection, recent exposure identification, and stronger correlation with blood than urine concentrations. Because cannabinoids are usually the most prevalent analytes in illicit drug testing, application of OF drug testing requires sufficient scientific data to support sensitive and specific OF cannabinoid detection. This review presents current knowledge of OF cannabinoids, evaluating pharmacokinetic properties, detection windows, and correlation with other biological matrices and impairment from field applications and controlled drug administration studies. In addition, onsite screening technologies, confirmatory analytical methods, drug stability, and effects of sample collection procedure, adulterants, and passive environmental exposure are reviewed. Delta-9-tetrahydrocannabinol OF concentrations could be >1000 µg/L shortly after smoking, whereas minor cannabinoids are detected at 10-fold and metabolites at 1000-fold lower concentrations. OF research over the past decade demonstrated that appropriate interpretation of test results requires a comprehensive understanding of distinct elimination profiles and detection windows for different cannabinoids, which are influenced by administration route, dose, and drug use history. Thus, each drug testing program should establish cut-off criteria, collection/analysis procedures, and storage conditions tailored to its purposes. Building a scientific basis for OF testing is ongoing, with continuing OF cannabinoids research on passive environmental exposure, drug use history, donor physiological conditions, and oral cavity metabolism needed to better understand mechanisms of cannabinoid OF disposition and expand OF drug testing applicability. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.

Drug concentrations in post-mortem femoral blood compared with therapeutic concentrations in plasma.

Abstract: Therapeutic drug concentrations measured in plasma are of limited value as reference intervals for interpretation in post-mortem (PM) toxicology. In this study, drug concentration distributions were studied in PM femoral venous blood from 57 903 Finnish autopsy cases representing all causes of death during an 11-year period. Cause-of-death information was obtained from death certificates issued by forensic pathologists. Median, mean, and upper percentile (90th, 95th, 97.5th) concentrations were calculated for 129 drugs. To illustrate how PM median concentrations relate to established therapeutic ranges in plasma, a PM blood/plasma relationship was calculated for each drug. Males represented 75% of the subjects and showed a lower median age (55 yrs) than females (59 yrs). In 43% of these cases, blood alcohol concentration was higher than 0.2‰, and the median was 1.8‰. Sixty-one (47%) of the 129 drugs showed a PM blood/plasma relationship of 1. For 22 drugs (17%), the relationship was 1. No marked correlation was found between the PM blood/plasma relationship and the volume of distribution (Vd). For 36 drugs, more than 10% of cases were fatal poisonings attributed to this drug as the main finding. These drug concentration distributions based on a large database provide a helpful reference not only to forensic toxicologists and pathologists but also to clinical pharmacologists in charge of interpreting drug concentrations in PM cases. © 2013 The Authors. Drug Testing and Analysis published by John Wiley & Sons, Ltd.

An investigation of the stability of emerging new psychoactive substances.

Abstract: The evolving nature of new psychoactive substances (NPS) - often referred to as 'legal highs', 'designer drugs' or 'bath salts' - presents an evolving challenge for toxicologists. Apart from the detection and identification of these compounds, further analytical challenges may arise from the presence of possible metabolites or degradation products which may have to be considered when devising an analytical strategy. Whilst there has been some stability research for some more established drugs of abuse and medicinal products, data on emerging NPS are less abundant. In order to address this need, 13 NPS (4-MEC, MDAI, methoxetamine, 5-MeO-DALT, 6-APB, MPA, 5-IAI, MDAT, 2-AI, AMT, 25C-NBOMe, AH-7921, 5-MAPB) were spiked in blood and plasma and kept at room temperature (20-23 °C). Detection and identification of the suspected breakdown products were carried out by high performance liquid chromatography with diode-array detection (HPLC-DAD), liquid chromatography-tandem mass spectrometry (LC-MS/MS), and ultra high performance liquid chromatography with high mass accuracy quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). 4-MEC became undetectable in blood within 14 days with a corresponding loss of 54% in plasma. A breakdown product was identified as dihydro-4-MEC which was also found in vivo in a case work sample. Storage of AMT led to a range of potential breakdown products which were also found in vivo. The remaining substances were found to be stable for at least 21 days in blood and plasma. This is the first time stability data have been published for these emerging substances and showed that additional compounds found during forensic casework were potential metabolites rather than instability products. In particular, presumptive metabolites of 25C-NBOMe and AH-7921 are presented. Copyright © 2013 John Wiley & Sons, Ltd. Language: en

Characterization of the designer benzodiazepine diclazepam and preliminary data on its metabolism and pharmacokinetics.

Abstract: Designer benzodiazepines, first offered in online shops selling 'research chemicals' in 2012, provide an attractive alternative to prescription-only benzodiazepines as they are readily available over the Internet at a low price. However, as data regarding pharmacokinetic parameters, metabolism, and detectability in biological fluids are limited, they present a challenge for forensic laboratories. Most recently, diclazepam (other names: 2-chlorodiazepam, Ro 5-3448 or 7-chloro-5-(2-chlorophenyl)-1-methyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one) emerged as a new compound in this class of drugs. In this paper, this new designer benzodiazepine was characterized utilizing nuclear magnetic resonance (NMR), gas chromatography-mass spectrometry (GC-MS) as well as liquid chromatography tandem mass spectrometry (LC-MS/MS) techniques. Furthermore, a self-experiment was performed to gain preliminary data on pharmacokinetic properties and to identify the main metabolites. For this purpose, one tablet of diclazepam (declared amount: 1 mg) was ingested by one of the authors, and serum as well as urine samples were collected for 14 and 21 days, respectively. Based on this study, diclazepam has an approximate elimination half-life of 42 h and is metabolized into the pharmacologically active benzodiazepines delorazepam, lorazepam, and lormetazepam which can be detected in urine for 6, 19, and 11 days, respectively, when applying the presented LC-MS/MS method. In serum, the consumption could be proven between 99 h post-intake targeting the parent compound and up to 10 days targeting the metabolite delorazepam. As immunochemical assays are applied for screening purposes quite often, detectability using this technique was assessed, especially since detection of low-dosed benzodiazepines can be sometimes problematic. However, only one of the utilized immunochemical assays was capable of detecting the intake of one tablet diclazepam, and the positive results were restricted to a few urine samples showing relatively high creatinine concentrations.

A methamphetamine analog (N,α‐diethyl‐phenylethylamine) identified in a mainstream dietary supplement

Abstract: Pharmaceuticals and banned substances have been detected in hundreds of purportedly natural supplements. Recently, several athletes have been disqualified from competition after testing positive for the methamphetamine analog N,α-diethyl-phenylethylamine (N,α-DEPEA). Athletes have claimed they unknowingly consumed the banned stimulant in workout supplements. Three samples from different lot numbers of Craze, a workout supplement, were analyzed to detect the presence and concentration of N,α-DEPEA. Two labs independently identified N,α-DEPEA in the supplement using ultra high performance liquid chromatography (UHPLC) coupled to an LTQ Orbitrap XL mass spectrometer and UHPLC-quadruple-time-of-flight mass (Q-TOF) spectrometer, respectively. The identity of N,α-DEPEA was confirmed using nuclear magnetic resonance and reference standards. Manufacturer recommended servings were estimated to provide 21 to 35 mg of N,α-DEPEA. N,α-DEPEA has never been studied in humans. N,α-DEPEA is a methamphetamine analog; however, its stimulant, addictive and other adverse effects in humans are entirely unknown. Regulatory agencies should act expeditiously to warn consumers and remove N,α-DEPEA from all dietary supplements. Copyright © 2013 John Wiley & Sons, Ltd. Language: en

Determination of 4-bromo-2,5-dimethoxy-N-[(2-methoxyphenyl)methyl]-benzeneethanamine (25B-NBOMe) in serum and urine by high performance liquid chromatography with tandem mass spectrometry in a case of severe intoxication

Abstract: We present a case of 4-bromo-2,5-dimethoxy-N-[(2-methoxyphenyl)methyl]-benzeneethanamine (25B-NBOMe), an N-benzyl phenethylamines derivative, intoxication and a high performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) method for detection and quantification of 25B-NBOMe.A 19-year-old male was found unresponsive with generalized grand mal seizure activity. On the second day of hospitalization, a friend admitted that the patient used ‘some unknown drug’ called 25B. Serum and urine collected were sent to the Virginia Commonwealth University Medical Center Toxicology Laboratory for analysis. An HPLC-MS/MS method for the identification and quantificationof 25B-NBOMe using 2-(2,5-dimethoxyphenyl)-N-(2-methoxybenzyl) ethanamine (25H-NBOMe)as the internal standard (ISTD)was developed. As this is a novel, single-case presentation, an assay validation was performed prior to testing to ensure the reliability of the analytical results. The serum and urine specimens were determined to contain 180 pg/ml and1900 pg/ml of 25B-NBOMe, respectively.

New phenethylamines in Europe.

Abstract: Sixteen phenethylamines are now included in Schedules I and II of the United Nations 1971 Convention on Psychotropic Substances. Most of the ring-substituted compounds are in Schedule I, whereas 2C-B, amphetamine, and methamphetamine are listed in Schedule II. Substances in Schedule IV (e.g. benzphetamine) are now regarded as obsolete pharmaceutical products. They all represent the 'old phenethylamines'. By 2013, nearly 100 illicit phenethylamines had been found in the European Union (EU). Of these, nine (MBDB, 4-MTA, PMMA, 2C-I, 2C-T-2, 2C-T-7, TMA-2, 5-IT and 4-MA) were submitted for risk assessment by the European Monitoring Centre for Drugs and Drug Addiction (EMCDDA). All except MBDB were recommended for EU-wide control. Of the 'new phenethylamines', 2C-B was the most commonly reported, but other 2C compounds were widespread. Many of the ring-substituted phenethylamines are described in the 1991 book PIHKAL. Many fused ring phenethylamines have appeared in the past few years; they include further benzofurans (e.g. 5-and 6-APB), indanylalkylamines (e.g. 5-IAP), dibenzofurans (e.g. 2C-B-FLY) and 2-aminopropylindoles (e.g.5-IT). The recent and rapid rise of phenethylamines with bulky N-substituents (e.g. 25I-NBOMe) has been particularly significant. Although not phenethylamines, it is notable that the thiophene bioisosteres of amphetamine and methamphetamine as well as certain conformationally-restricted variants (e.g. aminoindanes) have been found in recent drug seizures. In the United Kingdom Misuse of Drugs Act, most ring-substituted phenethylamines are either listed by name or are covered by generic definitions dating from 1977. In 2013, temporary generic legislation included a number of benzofurans, indanylalkylamines and certain 'NBOMe' compounds.

Changes in patterns of injecting drug use in Hungary: a shift to synthetic cathinones

Abstract: The spread of synthetic cathinone injecting is a new phenomenon observed in recent years in Hungary. Until 2010, when the first anecdotal reports on cathinone injecting appeared, injecting was associated with the use of heroin and amphetamine. In this paper we review available evidence of the changes in the drug market and a concurrent shift in patterns of injecting drug use that have been taking place in Hungary since 2010. Remarkable changes have been observed in police seizures data since 2010. While new psychoactive substances have appeared, the availability of heroin has dropped significantly. A qualitative study in 2011 revealed that these market changes correlate with changes in patterns of injecting drug use: decreasing heroin use and the appearance of mephedrone injecting were reported by treatment and needle and syringe programme (NSP) personnel. These changes are detectable in other routine epidemiological data collection systems in the following years as well (i.e. treatment, drug-related deaths, NSP clientele). Heroin-related treatment demand dropped, as did heroin-related mortality. Parallel to this, a growing number of clients appeared in treatment and in NSPs who were primarily injecting cathinones. The shift to cathinones can be observed in amphetamine and heroin injectors as well. Monitoring changes in patterns of injecting drug use are especially important because of the vulnerability of this drug-user population and the consequences of this high-risk route of drug administration. The realignment observed in Hungary is to be further investigated with regard to its determinants, changes in risk behaviour, and in treatment needs.

Hair analysis as a tool to evaluate the prevalence of synthetic cannabinoids in different populations of drug consumers

Abstract: Among the new psychoactive products, herbal mixtures containing synthetic cannabimimetics are likely the most abused worldwide In this study, a specific ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the detection of 23 synthetic cannabinoids in hair samples was developed in order to (1) expand the number of screened compounds, coherent with new substances emerging in the European territory, (2) evaluate their consumption on a large period of examination, and (3) evaluate the diffusion of cannabimimetics among different populations of drug consumers The method employs digestion of hair sample with NaOH followed by extraction with n-hexane/ethylacetate, and injection into the UHPLC-MS/MS system After validation, the method was applied to the analysis of 344 hair samples previously tested in our laboratory for the most common drugs Overall, 15 samples were found positive for at least one synthetic cannabinoid Coherent with previously published results, the present data show that young males, former or still active Cannabis consumers, represent the population most often involved in synthetic cannabimimetics consumption Several cases of poly-abuse were also determined The drug most frequently detected was JWH-073 (11 samples) generally at low concentration (mean 769 ± 144 pg/mg, median 19 pg/mg, range 16-505 pg/mg), followed by JWH-122 (8 samples, mean concentration: 544 ± 968 pg/mg, median 284 pg/mg, range 74-2800 pg/mg) Other detected drugs included JWH-250, JWH-081, JWH-018, JWH-210, JWH-019, and AM-1220 For several positive samples, the synthetic cannabinoid concentration was lower than 50 pg/mg, underlining the need for established cut-off values for discrimination between chronic consumption and occasional use (or external contamination) Language: en

Analysis of new classes of recreational drugs in sewage: Synthetic cannabinoids and amphetamine-like substances

Abstract: The analysis of sewage for the residues of commonly used illicit drugs has successfully been applied as a suitable approach for estimating community illicit drug use. The drug market is increasingly dynamic with new substances continually being marketed for recreational purposes. In this study, ultra-high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to simultaneously and quantitatively detect the exogenous biomarkers of new classes of recreational drugs in sewage collected from three different Norwegian cities (Oslo, Bergen, Hamar). The samples were screened for the presence of khat (d-norpseudoephedrine and cathinone), mephedrone, pseudoephedrine, 7-aminoflunitrazepam, para-methoxyamphetamine (PMA), para-methoxy-N-methylamphetamine (PMMA) and a selection of urinary metabolites of synthetic cannabinoids collectively termed ´Spice´ (5-3-1-naphthoyl-1H-indol-1-yl-pentanoic acid (JWH 018 N-pentanoic acid), 1-5-hydroxypentyl-1H-indol-3-ylnaphthalen-1-yl-methanone (JWH 018 N-5-hydroxypentyl), 4-3-1-naphthoyl-1H-indol-1-yl-butanoic acid (JWH 073 N-butanoic acid), 1-4-hydroxybutyl-1H-indol-3-ylnaphthalen-1-yl-methanone (JWH 073 N-4-hydroxybutyl), 1-5-hydroxypentyl-1H-indol-3-yl4-methylnaphthalen-1-yl-methanone (JWH 122 N-5-hydroxypentyl), 1-5-fluoro-4-hydroxypentyl-1H-indol-3-ylnaphthalen-1-ylmethanone (AM2201 N-4-hydroxypentyl), and 1-5-hydroxypentyl-1H-indol-3-yl4-methoxyphenyl-methanone (RCS-4 N-5-hydroxypentyl)). Limits of detection were 1 ng/L for amphetamine like compounds and 5 ng/L for the metabolites of synthetic cannabinoids while the limits of quantification were 3 and 15 ng/L, respectively. Three of the fourteen selected biomarkers (cathine, pseudoephedrine and the synthetic cannabinoid metabolite JWH-018 N-5-hydroxypentyl) were detected in sewage, alongside the illicit drugs (and/or metabolites) typically found in sewage (cocaine, benzoylecognine, methamphetamine, MDMA, and THC-COOH). The khat biomarker d-norpseudoephedrine was detected in Oslo sewage at a mean concentration of 93 ng/L that represents a daily load of 54 mg/day/1000 inhabitants. Pseudoephedrine was present at mean concentrations of between 27 and 67 ng/L representing normalized daily loads of between 10 (Hamar) and 24 mg/day/1000 inhabitants (Bergen). The daily normalized loads of JWH-018 N-5-hydroxypentyl were between 49 (Oslo) and 62 mg/day/1000 inhabitants (Hamar). This study demonstrates for the first time that sewage biomarker analysis can be applied to evaluate not only the use the traditional illicit drugs (cocaine, cannabis and amphetamines), but also the use of certain new synthetic drugs.

The rise of new psychoactive substance use in Australia.

Abstract: New Psychoactive Substances are now a feature of Australia's recreational drug market. Little is known, however, about the prevalence of use, the characteristics of people who use them and the relationship between the NPS and ecstasy markets. This study examined the prevalence and correlates of NPS use amongst a group of regular ecstasy users in Australia. Participants were recruited if they had used ecstasy at least six times in the previous six months, lived in a capital city and were over 16 years of age. Purposive sampling was used, recruiting through universities and colleges, word of mouth and street press. 654 participants were recruited in 2013. Respondents who had used an NPS in the past six months were compared to those who had not. NPS were used by 44% of the total sample. In 2013 2C-I (14%) and 2C-B (8%) were the most prevalent NPS. Respondents in the NPS group were younger and reported more frequent use of more types of drugs. They were also more likely to rate the purity of ecstasy as low relative to those in the no NPS group. NPS are now an established part of Australia's recreational drug scene and NPS with hallucinogenic effects are now used most commonly. Monitoring systems need to develop capacity to monitor this highly dynamic market.

Optimization, validation, and the application of liquid chromatography-tandem mass spectrometry for the analysis of new drugs of abuse in wastewater

Abstract: Recently it was demonstrated that the analysis of drugs of abuse (DOA) in wastewater can be used to track their use in communities This paper presents the optimization, validation, and application of an analytical procedure based on solid-phase extraction (SPE) and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) for the determination of some emerging drugs in influent wastewater The compounds of interest were the cathinone derivatives methylenedioxypyrovalerone (MDPV) and 4-methylmethcathinone (mephedrone (MEPH)), ketamine (KET) and its metabolites norketamine (NK) and dehydronorketamine (DHNK), as well as the major metabolite of cannabis, 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH) Except for DHNK, deuterated analogues were used as internal standards for quantification The use of a short C18 column (50 mm x 2 mm, 3 µm) allowed a good separation for the compounds analyzed in positive ionization mode (KET, NK, DHNK, MDPV, and MEPH) and for THC-COOH, which was analyzed in negative ionization mode, with a total run time of 13 min Sample preparation using SPE was optimized by comparing Oasis HLB and Oasis MCX sorbents The method was validated for each compound by assessing the following parameters: linearity, accuracy, precision, recovery, and relative process efficiency The lowest calibration level was considered as the lower limit of quantification (LLOQ) and was 5 ng/l for KET, NK, DHNK, MDPV, and MEPH and 20 ng/l for THC-COOH KET, and THC-COOH could be quantified in the majority of wastewater samples from three large wastewater treatment plants in Belgium The other compounds were below the LLOQ or could not be detected

Cross-reactivity of designer drugs, including cathinone derivatives, in commercial enzyme-linked immunosorbent assays

Abstract: Since the introduction of synthetic heroin, designer drugs have been increasing in prevalence in the United States drug market over the past few decades. Recently, 'legal highs' sold as 'bath salts' have become a household term for one such class of designer drugs. While a number of federal and state bans have been enacted, the abuse of these designer drugs still continues. Few assays have been developed for the comprehensive detection of such compounds, so it is important to investigate how they may or may not react in presumptive screens, i.e. pre-existing commercial immunoassays. In this experiment, 16 different ELISA reagents were evaluated to determine the cross-reactivity of 30 designer drugs, including 24 phenylethylamines (including 8 cathinone derivatives), 3 piperazines, and 3 tryptamines. Cross-reactivity towards most drugs was Language: en

Rapid and simple LC‐MS/MS screening of 64 novel psychoactive substances using dried blood spots

Abstract: The range of novel psychoactive substances (NPS) including phenethylamines, cathinones, piperazines, tryptamines, etc. is continuously growing. Therefore, fast and reliable screening methods for these compounds are essential and needed. The use of dried blood spots (DBS) for a fast straightforward approach helps to simplify and shorten sample preparation significantly. DBS were produced from 10 µl of whole blood and extracted offline with 500 µl methanol followed by evaporation and reconstitution in mobile phase. Reversed-phase chromatographic separation and mass spectrometric detection (RP-LC-MS/MS) was achieved within a run time of 10 min. The screening method was validated by evaluating the following parameters: limit of detection (LOD), matrix effect, selectivity and specificity, extraction efficiency, and short-term and long-term stability. Furthermore, the method was applied to authentic samples and results were compared with those obtained with a validated whole blood method used for routine analysis of NPS. LOD was between 1 and 10 ng/ml. No interference from matrix compounds was observed. The method was proven to be specific and selective for the analytes, although with limitations for 3-FMC/flephedrone and MDDMA/MDEA. Mean extraction efficiency was 84.6 %. All substances were stable in DBS for at least a week when cooled. Cooling was essential for the stability of cathinones. Prepared samples were stable for at least 3 days. Comparison to the validated whole blood method yielded similar results. DBS were shown to be useful in developing a rapid screening method for NPS with simplified sample preparation.

A proteomic evaluation of the effects of the pharmaceuticals diclofenac and gemfibrozil on marine mussels (Mytilus spp.): evidence for chronic sublethal effects on stress-response proteins.

Abstract: Human pharmaceuticals (e.g. the lipid regulator gemfibrozil and the non-steroidal anti-inflammatory drug diclofenac) are an emerging environmental threat in the aquatic environment. This study aimed to evaluate sublethal effects of these two commonly found pharmaceuticals on the protein profiles of marine mussels (Mytilus spp.). Mytilus spp. was exposed to environmentally relevant and elevated concentrations (1 and 1000 µg/l respectively) of both drugs for 14 days. In addition, mussels were maintained for seven days post treatment to examine the potential of blue mussels to recover from such an exposure. Differential protein expression signatures (PES) in the digestive gland of mussels were obtained using two-dimensional gel electrophoresis after 7, 14, and 21 days of exposure. Twelve spots were significantly increased or decreased by gemfibrozil and/or diclofenac, seven of which were successfully identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. These proteins were involved in energy metabolism, oxidative stress response, protein folding, and immune responses. Changes in the PES over time suggested that mussels were still experiencing oxidative stress for up to seven days post exposure. In addition, a suite of biomarkers comprising glutathione transferase, lipid peroxidation, and DNA damage were studied. An oxidative stress response was confirmed by biomarker responses. To our knowledge, this is the first investigation using proteomics to assess the potential effects of human pharmaceuticals on a non-target species in an environmentally-relevant model. The successful application of this proteomic approach supports its potential use in pollution biomonitoring and highlights its ability to aid in the discovery of new biomarkers.

Identification and quantification of synthetic cannabinoids in 'spice-like' herbal mixtures: a snapshot of the German situation in the autumn of 2012.

Abstract: Synthetic compounds mimicking cannabis-like effects are a recent trend. Currently, these so-called synthetic cannabinoids are the largest and fastest growing class of newly appearing designer drugs. Many national authorities are continuously adapting their regulations to keep pace with the permanently changing variety of compounds. We have analyzed eight herbal smoking blends containing synthetic cannabinoids. Altogether, nine compounds could be identified, namely AM-2201, AM-2201-pMe (MAM-2201), AM-1220, AM-1220-azepane, UR-144, XLR-11, JWH-122-pentenyl, AM-2232, and STS-135. Newly appearing compounds were isolated by column chromatography and their structures elucidated by 1D- and 2D-nuclear magnetic resonance (NMR) experiments. In addition, the compounds were investigated by electron ionization-mass spectrometry (EI-MS) and electrospray ionization-tandem mass spectrometry (ESI-MS/MS) to complete the physicochemical dataset. Based on the purified compounds a universal gas chromatography-mass spectrometry (GC-MS) method was developed for the identification and quantification of these compounds in commercial smoking blends. By applying this method, up to five different compounds could be found in such products showing total concentrations from 72 to 303 mg/g smoking blend while individual compounds ranged from 0.4 to 303 mg/g. 1H NMR spectra of the chiral compounds AM-1220 and its azepane-isomer recorded in the presence of 1 equivalent of (R)-(+)-α-methoxy-α-trifluoromethylphenylacetic acid (MTPA, Mosher's acid) showed them to be racemic mixtures. Copyright © 2013 John Wiley & Sons, Ltd.

Determination of human insulin and its analogues in human blood using liquid chromatography coupled to ion mobility mass spectrometry (LC‐IM‐MS)

Abstract: The qualitative and quantitative determination of insulin from human blood samples is an emerging topic in doping controls as well as in other related disciplines (e.g. forensics). Beside the therapeutic use, insulin represents a prohibited, performance enhancing substance in sports drug testing. In both cases accurate, sensitive, specific, and unambiguous determination of the target peptide is of the utmost importance. The challenges concerning identifying insulins in blood by liquid chromatography coupled to ion mobility mass spectrometry (LC-IM-MS) are detecting the basal concentrations of approximately 0.2 ng/mL and covering the hyperinsulinaemic clamps at > 3 ng/mL simultaneously using up to 200 μL of plasma or serum. This is achieved by immunoaffinity purification of the insulins with magnetic beads and subsequent separation by micro-scale liquid chromatography coupled to ion mobility / high resolution mass spectrometry. The method includes human insulin as well as the synthetic or animal analogues insulin aspart, glulisine, glargine, detemir, lispro, bovine, and porcine insulin. The method validation shows reliable results considering specificity, limit of detection (0.2 ng/mL except for detemir: 0.8 ng/mL), limit of quantification (0.5 ng/mL for human insulin), precision (CV 0.99), recovery, accuracy (>90%), robustness (plasma/serum), and ion suppression. For quantification of human insulin a labelled internal standard ([[2H10]-LeuB6,B11,B15,B17] ‒ human Insulin) is introduced. By means of the additional ion mobility separation of the different analogues, the chromatographic run time is shortened to 8 min without losing specificity. As proof-of-concept, the procedure was successfully applied to different blood specimens from diabetic patients receiving recombinant synthetic analogues Copyright © 2014 John Wiley & Sons, Ltd.

Validation of the only commercially available immunoassay for synthetic cathinones in urine: Randox Drugs of Abuse V Biochip Array Technology

Abstract: Deterrence of synthetic cathinone abuse is hampered by the lack of a high-throughput immunoassay screen. The Randox Drugs of Abuse V (DOA-V) Biochip Array Technology contains two synthetic cathinone antibodies: Bath Salt I (BSI) targets mephedrone/methcathinone and Bath Salt II (BSII) targets 3',4'-methylenedioxypyrovalerone (MDPV)/3',4'-methylenedioxy-α-pyrrolidinobutiophenone (MDPBP). We evaluated DOA-V synthetic cathinones performance and conducted a full validation on the original assay with calibrators reconstituted in water, and the new assay with calibrators prepared in lyophilized urine; both utilized the same antibodies and were run on the fully automated Evidence® Analyzer. We screened 20 017 authentic military urine specimens and confirmed positives by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for 28 synthetic cathinones. Limits of detection (LOD) for the original and new assays were 0.35 and 0.18 (BSI), and 8.5 and 9.2 µg/L (BSII), respectively. Linearity was acceptable (R(2) >0.98); however, a large negative bias was observed with in-house prepared calibrators. Intra-assay imprecision was <20% BSI-II, while inter-assay imprecision was 18-42% BSI and <22% BSII. Precision was acceptable for Randox controls. Cross-reactivities of many additional synthetic cathinones were determined. Authentic drug-free negative urine pH <4 produced false positive results for BSI (6.3 µg/L) and BSII (473 µg/L). Oxidizing agents reduced BSI and increased BSII results. Sensitivity, specificity, and efficiency of 100%, 52.1%, and 53.0% were obtained at manufacturer's proposed cut-offs (BSI 5 µg/L, BSII 30 µg/L). Performance improved if cut-off concentrations increased (BSI 7.5 µg/L, BSII 40 µg/L); however, there were limited confirmed positive specimens. Currently, this is the first and only fully validated immunoassay for preliminary detection of synthetic cathinones in urine. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

A cocktail of synthetic stimulants found in a dietary supplement associated with serious adverse events

Abstract: Food supplements are regularly found to contain pharmacologically active substances. Recently, the food supplement Dexaprine was removed from the Dutch market because it was associated with severe adverse events. Reports to the Dutch Poisons Information Center (DPIC) showed that ingestion of as little as half a tablet caused several cases of nausea, agitation, tachycardia, and palpitations and even one case of cardiac arrest. The remaining tablets of four patients were sent in by different healthcare professionals. Analysis by ultra-performance liquid chromatography quadrupole time of flight mass-spectrometry (UPLC-QTOF-MS) confirmed the presence of synephrine, oxilofrine, deterenol, yohimbine, caffeine, and theophylline. Two more compounds were found which were tentatively identified as β-methyl-β-phenylethylamines. This incident is only the next in a series of similar incidents involving dietary supplements with (undeclared) active substances that are either unsafe or have no known safety profile.

Hair analysis for THCA-A, THC and CBN after passive in vivo exposure to marijuana smoke.

Abstract: Condensation of marijuana smoke on the hair surface can be a source of an external contamination in hair analysis and may have serious consequences for the person under investigation. Δ9-tetrahydrocannabinolic acid A (THCA-A) is found in marijuana smoke and in hair analysis, but is not incorporated into the hair through the bloodstream. Therefore it might be a promising marker for external contamination of hair and could facilitate a more accurate interpretation of analytical results. In this study, three participants were exposed to the smoke of one joint every weekday over three weeks. Inhalation was excluded by an alternative breathing source. Hair samples were obtained up to seven weeks after the last exposure and analyzed for THCA-A, Δ9-tetrahydrocannabinol (THC) and cannabinol (CBN) by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Additionally 30 hair samples from various regions of the head were obtained seven weeks after the exposure from one participant. The obtained results show that the degree of contamination depends on the hair length, with longer hair resulting in higher THC and CBN concentrations (1300 pg/mg and 530 pg/mg at the end of the exposure period) similar to the ones typically found after daily cannabis consumption. THCA-A could be detected in relatively low concentrations. Analysis of the distribution of the contamination showed that the posterior vertex region was affected most. The relatively low THCA-A concentrations in the samples suggest that most of the THCA-A found in forensic hair samples is not caused by sidestream marijuana smoke, but by other sources. Language: en

Intolerability of cobalt salt as erythropoietic agent

Abstract: Unfair athletes seek ways to stimulate erythropoiesis, because the mass of haemoglobin is a critical factor in aerobic sports. Here, the potential misuse of cobalt deserves special attention. Cobalt ions (Co(2+) ) stabilize the hypoxia-inducible transcription factors (HIFs) that increase the expression of the erythropoietin (Epo) gene. Co(2+) is orally active, easy to obtain, and inexpensive. However, its intake can bear risks to health. To elaborate this issue, a review of the pertinent literature was retrieved by a search with the keywords 'anaemia', 'cobalt', 'cobalt chloride', 'erythropoiesis', 'erythropoietin', 'Epo', 'side-effects' and 'treatment', amongst others. In earlier years, cobalt chloride was administered at daily doses of 25 to 300 mg for use as an anti-anaemic agent. Co(2+) therapy proved effective in stimulating erythropoiesis in both non-renal and renal anaemia, yet there were also serious medical adverse effects. The intake of inorganic cobalt can cause severe organ damage, concerning primarily the gastrointestinal tract, the thyroid, the heart and the sensory systems. These insights should keep athletes off taking Co(2+) to stimulate erythropoiesis.

Characterizing metabolites and potential metabolic pathways for the novel psychoactive substance methoxetamine

Abstract: The classic approach of controlled volunteer studies to study drug metabolism is dif ficult or impossible to undertake for novel psy-choactive substances (NPS), as there is generally very l imited information available to allow appropriate dose finding and safety. Aviable and powerful alternative is the identification and characterization of phase I and II metabolites of such drugs by examiningthe concordance of data gathered from analysis of microsomal i ncubates with that from analysis of specimens collected fromindividuals with analytically confirmed use of NPS. Liquid chromatography-high resolution mass spectrometry provides the abilitytoreliablyidentifysuchmetabolites.Weusedthistechniquehere tostudythemetabolismoftheketamineanaloguemethoxetamine.A large number of metabolites were identi fied in the in vitro studies including normethoxetamine, O-desmethylmethoxetamine,dihydromethoxetamine, dehydromethoxetamine and several structural isomers of hydroxymethoxetamine andhydroxynormethoxetamine. pH dependent liquid-liquid extraction was used to discriminate phenolic from alcoholic metab-olites. Phase II glucuronide conjugates included those ofO-desmethylmethoxetamine,O-desmethylnormethoxetamine andO-desmethylhydroxymethoxetamine. The majority of these phase I and II metabolites were confirmed to be present in urinecollected from three individuals presentingwithacutemethoxetaminetoxicity. Copyright © 2013 John Wiley & Sons, Ltd.Additional supporting information may be found in the online version of this article at the publisher’s web site.Keywords: methoxetamine; high resolution mass spectrometry; new psychoactive substance; metabolism