Enantiomeric profiling of amphetamines in wastewater using chiral derivatisation with gas chromatographic-tandem mass spectrometric detection.

Enantiomeric profiling can supplement wastewater-based epidemiology (WBE) by providing additional information on drug origin (licit or illicit), improving consumption estimates, i.e., differentiating between disposal and consumption, and offering an insight into the potency of drugs available on the illicit drug market. We report on the enantiomeric profiling of amphetamines in wastewater using R(-)-α-methoxy-α-(trifluoromethyl) phenylacetyl chloride (R-MTPCl), a chiral derivatising agent and GC-MS/MS. The method performed well when evaluated against the SANTE/12682/2019 guidelines in terms of recovery (81-99%), accuracy (99-111%), repeatability (1-8%RSD) and linearity (LOQ-1000 ng/mL). The LOD and LOQ were 120 ng/L and 400 ng/L, respectively. The method was applied to samples of raw wastewater from two Slovene municipalities with unusual levels of amphetamines: Ljubljana (LJ1) and Velenje (VE1). LJ1 had an anomalously high mass load of MDMA (3,4-methylenedioxymethamphetamine) identified during SCORE 2020, and VE1 is a representative sample of the consistently high mass load of amphetamine. A second Ljubljana sample (LJ2) was chosen as a representative sample. The presence of racemic MDMA (EF = 0.511) in LJ1 indicated the disposal of the unused drug into the sewer, while the enrichment of R-MDMA (EF = 0.666) in the combined extract sample from Ljubljana (LJ2) indicated consumption. In the case of Velenje and Ljubljana, it is impossible to distinguish between the direct disposal and consumption of amphetamine and methamphetamine. Also, since amphetamine/methamphetamine-based prescription medications are unavailable in Slovenia, racemic amphetamine in VE1 (EF = 0.514) and LJ2 (EF = 0.459) indicate racemic and the more potent S-amphetamine are sold on the illicit drug market. Only S-methamphetamine was detected in wastewater (LJ2: EF = 0), indicating the presence of only the more potent S-methamphetamine on the illicit drug market. Overall, enantiomeric profiling provided useful information on amphetamine residues. In addition, chiral derivatisation can be a cost-effective alternative to using chiral chromatographic columns for the enantiomeric profiling of amphetamines in wastewater.

Revealing Metabolic Perturbation Following Heavy Methamphetamine Abuse by Human Hair Metabolomics and Network Analysis.

Methamphetamine (MA) is a highly addictive central nervous system stimulant. Drug addiction is not a static condition but rather a chronically relapsing disorder. Hair is a valuable and stable specimen for chronic toxicological monitoring as it retains toxicants and metabolites. The primary focus of this study was to discover the metabolic effects encompassing diverse pathological symptoms of MA addiction. Therefore, metabolic alterations were investigated in human hair following heavy MA abuse using both targeted and untargeted mass spectrometry and through integrated network analysis. The statistical analyses (t-test, variable importance on projection score, and receiver-operator characteristic curve) demonstrated that 32 metabolites (in targeted metabolomics) as well as 417 and 224 ion features (in positive and negative ionization modes of untargeted metabolomics, respectively) were critically dysregulated. The network analysis showed that the biosynthesis or metabolism of lipids, such as glycosphingolipids, sphingolipids, glycerophospholipids, and ether lipids, as well as the metabolism of amino acids (glycine, serine and threonine; cysteine and methionine) is affected by heavy MA abuse. These findings reveal crucial metabolic effects caused by MA addiction, with emphasis on the value of human hair as a diagnostic specimen for determining drug addiction, and will aid in identifying robust diagnostic markers and therapeutic targets.

Liquid chromatography-high resolution mass spectrometry for the determination of three cannabinoids, two (-)-trans-Δ9-tetrahydrocannabinol metabolites, and six amphetamine-type stimulants in human hair.

Since cannabis and amphetamine-type stimulants (ATS) are drugs of abuse used alone and concurrently worldwide, it is important to analyze them simultaneously. However, there are no reports of analytical method for the simultaneous extraction of these compounds and metabolites in hair samples of suspected drug abusers, due to the different chemical properties of acidic and lipophilic cannabinoids, and basic and hydrophilic ATS. We developed a liquid chromatography-high resolution mass spectrometry (LC-HRMS) method for the quantification of cannabidiol (CBD), cannabinol (CBN), (-)-trans-Δ9-tetrahydrocannabinol (THC), THC metabolites such as 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) and THC-COOH-glucuronide (THC-COOH-glu), and six ATS of amphetamine, methamphetamine, phentermine, methylenedioxyamphetamine, methylenedioxymethamphetamine, and methylenedioxyethylamphetamine in the least amounts of human hair samples. The pulverized hair samples (10 mg) were extracted with 1 mL of 0.5% formic acid in methanol three times, and the supernatants were evaporated in a stream of nitrogen gas. The residue was dissolved and the aliquot was analyzed by LC-HRMS using positive electrospray ionization and the parallel reaction monitoring mode. The lower limits of quantitation were 0.1 pg mg-1 for THC-COOH and THC-COOH-glu, 4 pg mg-1 for CBD, CBN, and THC, and 10 pg mg-1 for the six ATS. Linearity, accuracy, precision, matrix effect, recovery, and post-preparation stability for the 11 analytes were fully validated. The present method was successfully applied to the determination of 11 analytes in hair samples of 10 suspected drug abusers.

Prevalence and factors associated to the use of illicit drugs and psychotropic medications among brazilian undergraduates

The aim of this study was to verify the prevalence of use of psychoactive substances (PS) and its associated factors in undergraduate students of a university in southern Brazil. The study was carried out with 830 undergraduate students in the year 2016. The individuals answered a self-administered questionnaire about the PS and its prevalence of daily use, in the last 30 days or at any time of their lives, as well as socioeconomic conditions and academic variables. Caffeine-based energy drinks was the most consumed psychoactive substance (96.3%) among undergraduates in the last 30 days, followed by alcohol (64.0%). Among the illicit drugs most consumed in the last 30 days was marijuana (17.3%), while anxiolytics and amphetamines were the most prevalent psychoactive medicaments in the last 30 days. The prevalence of lifetime illicit drugs used by these students was 41.5%, where we highlight besides marijuana (38.6%) the high consumption of cocaine (7.8%), ecstasy (9.3%) and solvents. Socioeconomic and demographic factors such as gender, have children, religion, and financial background as well as academic variables were associated to recent consumption of these substances. This study concluded there is a high prevalence of use of PS among the undergraduate students, including illicit drugs.

Characterisation of aqueous waste produced during the clandestine production of amphetamine following the Leuckart route utilising solid-phase extraction gas chromatography-mass spectrometry and capillary electrophoresis with contactless conductivity detection.

Chemical waste from the clandestine production of amphetamine is of forensic and environmental importance due to its illegal nature which often leads to dumping into the environment. In this study, 27 aqueous amphetamine waste samples from controlled Leuckart reactions performed in Germany, the Netherlands, and Poland were characterised to increase knowledge about the chemical composition and physicochemical characteristics of such waste. Aqueous waste samples from different reaction steps were analysed to determine characteristic patterns which could be used for classification. Conductivity, pH, density, ionic load, and organic compounds were determined using different analytical methods. Conductivity values ranged from 1 to over 200 mS/cm, pH values from 0 to 14, and densities from 1.0 to 1.3 g/cm3 . A capillary electrophoresis method with contactless conductivity detection (CE-C4 D) was developed and validated to quantify chloride, sulphate, formate, ammonium, and sodium ions which were the most abundant ions in the investigated waste samples. A solid-phase extraction sample preparation was used prior to gas chromatography-mass spectrometry analysis to determine the organic compounds. Using the characterisation data of the known samples, it was possible to assign 16 seized clandestine waste samples from an amphetamine production to the corresponding synthesis step. The data also allowed us to draw conclusions about the synthesis procedure and used chemicals. The presented data and methods could support forensic investigations by showing the probative value of synthesis waste when investigating the illegal production of amphetamine. It can also act as starting point to develop new approaches to tackle the problem of clandestine waste dumping.

Avaliação da presença de cocaína e anfetamina em amostras de sangue post mortem e de indivíduos vivos, utilizando técnica de microextração em fase líquida (HF-LPME) / Amphetamine, cocaine and tetrahydrocannabinol evaluation in blood samples of living people and post mortem blood samples using microextraction technique in liquid phase (HF-LPME)

Estima-se atualmente que mais de 5% da população mundial vem fazendo uso recreativo de algum tipo de substância psicoativa, sendo que o direito a esse uso é tema recorrente da sociedade contemporânea. Por apresentar riscos associados à saúde e a segurança das populações, o uso abusivo dessas substâncias tem instigado a toxicologia social na busca de respostas, com as quais se possa caracterizar, analisar e gerenciar esses riscos. Drogas de grande consumo no Brasil são a anfetamina, cocaína e Cannabis sativa. Esta tese desenvolveu uma nova metodologia para detectar e quantificar anfetamina, cocaína e tetrahidrocanabinol em sangue total, com uso de microextração em fase líquida via fibra de polipropileno (HF-LPME), seguida de cromatografia gasosa acoplada a espectrometria de massa (GC-MS). Trata-se de uma técnica que apresenta vantagens sobre as tradicionais, uma vez que demanda quantidades menores de solvente orgânico, diminuindo riscos e custos de processo. Também propôs um estudo com a aplicação dos métodos em 69 amostras de sangue de vivos e de post mortem, as quais foram obtidas por convênio com a superintendência da polícia técnica científica de São Paulo (SPTC/SP). Os métodos desenvolvidos foram validados de acordo com diretrizes internacionais de interesse forense. Como resultado da validação, os métodos desenvolvidos se mostraram precisos e exatos para anfetamina e cocaína. O limite de detecção da cocaína foi de 5 ng . mL-1 e o limite de quantificação de 10 ng . mL-1. Quanto a anfetamina, os limites de detecção e de quantificação foram de 5 ng . mL-1. A técnica de HF-LPME não foi aplicável ao tetraidrocanabinol (Δ9-THC). Como resultado da análise das amostras, 40% delas apresentaram resultados positivos para cocaína. Desses positivos, 35% foram oriundos das matrizes de sangue de vivos e 64% oriundos de sangue post mortem. Nenhuma delas apresentou resultado quantificável para anfetamina

It is currently estimated that more than 5% of the world's population has been doing recreational use of some kind of psychoactive substances and the legal right to such use is a recurring theme debated by contemporary society. Due to the risks associated with populations health and safety, the abusive use of these substances has been instigating by social toxicology to search for answers to characterize, analyze and manage these risks. Drugs of great consumption in Brazil are, amphetamine cocaine and marijuana. This thesis proposes to develop a new methodology to detect and quantify psychoactive drugs in whole blood with the use of liquid phase microextraction by polypropylene fiber (HFLPME), followed by gas chromatography coupled to mass spectrometry (GC-MS). It is a technique that presents advantages compared with traditional ones, because of the smaller amounts demands of organic solvent, reducing risks and process costs. It also proposes a study with 69 blood samples taken from living persons and post mortem blood samples, which were obtained by agreement with the Superintendency of São Paulo Scientific Technical Police (SPTC / SP). The methods developed were validated according to international guidelines of forensic interest. As a result of the validation, the methods developed were precise and accurate for amphetamine and cocaine. The limit of cocaine detection was 5 ng . mL-1 and the limit of quantification was 10 ng . mL-1. As for amphetamine, the limits of detection and quantification were 5 ng . mL-1. The HF-LPME technique was not applicable to tetrahydrocannabinol (Δ9-THC). As a result of the sample analysis, 40% of them presented positive results for cocaine. Of these, 35% were from blood samples taken from living persons and 64% from the post mortem blood samples. None of the samples presented quantifiable results for amphetamine

A Norwegian study of the suitability of hair samples in epidemiological research of alcohol, nicotine and drug use.

A feasibility study was performed to examine the effectiveness of hair testing in determining the prevalence of drug use in a young adult population. The study included 200 randomly selected young adults in Norway. It was designed to make the collection, preparation and analysis of the samples as little resource demaning as possible. Full anonymity was provided for the participants. In total, 23.5% of the samples were positive for one or more substances (14.5%, excluding the nicotine metabolite cotinine). Of the samples, 5% were positive for at least one illegal drug, 9.5% for a medicinal drug, 11.5% for cotinine and 2.5% for the alcohol metabolite ethyl glucuronide. The preliminary findings suggest that the study protocol used to collect and analyze the samples was unable to produce results that could be generalized to the young adult population in Norway. Analysis of hair samples may underestimate the use of cannabis, alcohol, amphetamine and methamphetamine. It may, however, be done to estimate cocaine and general drug use if a sample-collection procedure different from that described in our study is used and includes information about hair length, sample length, length from the scalp, cosmetic treatment, washing and whether the samples always get washed/decontaminated prior to analysis.

Determination of amphetamine-type stimulants in oral fluid by solid-phase microextraction and gas chromatography-mass spectrometry.

A method for the simultaneous identification and quantification of amphetamine (AMP), methamphetamine (MET), fenproporex (FEN), diethylpropion (DIE) and methylphenidate (MPH) in oral fluid collected with Quantisal™ device has been developed and validated. Thereunto, in-matrix propylchloroformate derivatization followed by direct immersion solid-phase microextraction and gas chromatography-mass spectrometry were employed. Deuterium labeled AMP was used as internal standard for all the stimulants and analysis was performed using the selected ion monitoring mode. The detector response was linear for the studied drugs in the concentration range of 2-256 ng mL(-1) (neat oral fluid), except for FEN, whereas the linear range was 4-256 ng mL(-1). The detection limits were 0.5 ng mL(-1) (MET), 1 ng mL(-1) (MPH) and 2 ng mL(-1) (DIE, AMP, FEN), respectively. Accuracy of quality control samples remained within 98.2-111.9% of the target concentrations, while precision has not exceeded 15% of the relative standard deviation. Recoveries with Quantisal™ device ranged from 77.2% to 112.1%. Also, the goodness-of-fit concerning the ordinary least squares model in the statistical inference of data has been tested through residual plotting and ANOVA. The validated method can be easily automated and then used for screening and confirmation of amphetamine-type stimulants in drivers' oral fluid.

HIV-negative drug addict diagnosed with AIDS and tuberculosis at autopsy: a case report and brief review of literature.

Human Immunodeficiency Virus (HIV) infection is a true plague and a major health concern globally. It is one of the most significant pandemics in recorded history. Despite worldwide efforts to fight the pandemic, and now with the re-emergence of tuberculosis, those clinicians, personnel performing autopsies and medical caregivers are again at risk in the work place, especially in developing countries. We describe a case where a drug abuser, whose addiction was concealed by his parents, died in hospital. He was tested HIV-negative there. A medical tangle ensued and forensic autopsy was carried out. Autopsy confirmed he was an intravenous drug addict and had tuberculosis. Post-mortem blood was positive for HIV antibodies and he was diagnosed with AIDS. Due to social stigmas, lack of knowledge or inefficient medical laboratory procedures etc, such type of cases can become a hazard to those attending the sick and to autopsy pathologists alike. We provide the case description, autopsy findings and review of pertinent literature.

Rapid and simple determination of psychotropic phenylalkylamine derivatives in human hair by gas chromatography-mass spectrometry using micro-pulverized extraction.

A gas chromatography-mass spectrometric (GC-MS) method was developed and validated for the determination of five psychotropic phenylalkylamine derivatives (amphetamine, AP; methamphetamine, MA; 3,4-methylenedioxyamphetamine, MDA; 3,4-methylenedioxymethamphetamine, MDMA; norketamine, NKT) in human hair. Hair samples (10mg) were washed with distilled water and acetone, mechanically pulverized for 1.5 min with a bead mill, and then incubated in 1 mL of methanol under ultrasonication at 50 degrees C for 1h. The resulting solutions were evaporated to dryness, derivatized using heptafluorobutyric anhydride (HFBA) at 50 degrees C for 30 min, and analyzed by GC-MS. The linear ranges were 0.1-20.0 ng/mg for AP and MA and 0.05-20.0 ng/mg for MDA, MDMA, and NKT, with the coefficients of determination (r(2)>0.9982). The intra-day and inter-day precisions were within 11.5% and 12.8%, respectively. The intra-day and inter-day accuracies were -4.1% to 5.8% and -6.6% to 4.2%, respectively. The limits of detections (LODs) for each compound were lower than 0.028 ng/mg. The recoveries were in the range of 78.9-101.2%. Based on these results, the method proved to be effective for the rapid and simple determination of phenylalkylamine derivatives in hair specimens.

Determination of 4-hydroxy-3-methoxymethamphetamine as a metabolite of methamphetamine in rats and human liver microsomes using gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry.

The aim of this study was to determine whether methamphetamine (MA) is metabolized to 4-hydroxy-3-methoxymethamphetamine (HMMA), which is known as the main metabolite of 3,4-methylenedioxymethamphetamine (MDMA). After MA was intravenously administered to rats, the plasma, urine, and bile were collected periodically. HMMA together with MA and its main metabolites, amphetamine and 4-hydroxymethamphetamine, were detected in the rat plasma, urine, and bile by gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry. In addition, HMMA was produced when MA was incubated with human liver microsomes. HMMA may be produced as a metabolite of MA when humans have consumed MA, although the amount of HMMA would be small compared with that of MA, amphetamine, or 4-hydroxymethamphetamine. The results of the present study will be helpful in determining the type of drug used.

Causes and manners of death among users of heroin, methadone, amphetamine, and cannabis in relation to postmortem chemical tests for illegal drugs.

A 12-year medicolegal investigation of deceased illegal drug users (ILDU) in Stockholm, Sweden, classified on the basis of postmortem chemical tests, showed noticeable variations in causes and manners of death as well as in the distribution of suicide methods. This study offers objective information about connection between the postmortem findings of illegal drugs and the causes and manners of death of their users. However, further studies, comparing prevalence of drug use in general population and at the postmortem tests, are needed for more detailed elucidation of this connection.

Simultaneous determination of amphetamine-type stimulants and cannabinoids in fingernails by gas chromatography-mass spectrometry.

A gas chromatography-mass spectrometric (GC-MS) method was developed and validated for the simultaneous detection and quantification of four amphetamine-type stimulants (amphetamine (AP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA)) and two cannabinoids (Delta9-tetrahydrocannabinol (Delta9-THC) and 11-nor-Delta9-tetrahydrocannabinol-9-carboxylic acid (THCCOOH)) in fingernails. Fingernail clippings (30 mg) were washed with distilled water and methanol, and then incubated in 1.0 M sodium hydroxide at 95 degrees C for 30 min. The compounds of interest were isolated by liquid-liquid extraction followed by derivatization with N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) at 70 degrees C for 15 min. The derivatized compounds were analyzed by GC-MS in the selective ion monitoring (SIM) mode. The linear ranges were 0.1-15.0 ng/mg for AP, 0.2-15.0 ng/mg for MDA, Delta9-THC and THCCOOH, and 0.2-30.0 ng/mg for MA and MDMA, with good correlation coefficients (r2 > 0.9991). The intra-day, inter-day, and inter-person precisions were within 10.6%, 6.3%, and 5.3%, respectively. The intra-day, inter-day and inter-person accuracies were between -6.1 and 5.0%, -6.2 and 5.7%, and -6.4 and 5.6%, respectively. The limits of detection (LODs) and quantification (LOQs) for each compound were lower than 0.056 and 0.2 ng/mg, respectively. The recoveries were in the range of 74.0-94.8%. Positive GC-MS results were obtained from specimens of nine suspected MA or cannabis abusers. The concentration ranges of MA, AP, and THCCOOH were 0.10-1.41, 0.12-2.64, and 0.20 ng/mg, respectively. Based on these results, the method proved to be effective for the simultaneous qualification and quantification of amphetamine-type stimulants and cannabinoids in fingernails.

Screening for drugs in oral fluid: illicit drug use and drug driving in a sample of Queensland motorists.

Police Services in a number of Australian states have indicated random roadside drug testing will be implemented to target drug driving. This paper outlines research conducted to provide an estimate of the prevalence of drug driving in a sample of Queensland drivers. Oral fluid samples were collected from 781 drivers who volunteered to participate at Random Breath Testing (RBT) sites in a large Queensland regional area. Illicit substances tested for included cannabis (delta 9 tetrahydrocannibinol [THC]), amphetamine type substances, heroin and cocaine. Drivers also completed a self-report questionnaire regarding their drug-related driving behaviour. Samples that were drug-positive at initial screening were sent to a government laboratory for confirmation. Oral fluid samples from 27 participants (3.5%) were confirmed positive for at least one illicit substance. The most common drugs detected in oral fluid were cannabis (delta 9 THC) (n = 13) followed by amphetamine type substances (n = 11). A key finding was that cannabis was also confirmed as the most common self-reported drug combined with driving and that individuals who tested positive to any drug through oral fluid analysis were also more likely to report the highest frequency of drug driving. Furthermore, a comparison between drug vs drink driving detection rates for the study period revealed a higher detection rate for drug driving (3.5%) vs drink driving (0.8%). This research provides evidence that drug driving is relatively prevalent on Queensland Roads. The paper will further outline the study findings and present possible directions for future drug driving research.

Time-resolved hair analysis of MDMA enantiomers by GC/MS-NCI.

The aim of the study was to determine the enantioselective disposition of 3,4-methylenedioxymethamphetamine (MDMA) and other amphetamine-type stimulants (ATS) in segmented hair specimens of self-declared ecstasy abusers, who took part in a double-blind placebo-controlled six-way crossover study during approximately 7 weeks, during which they received a 75 and a 100 mg dose of racemic MDMA twice. Hair specimens were washed and cut into pieces of 2 cm length. After digestion and solid phase extraction, the enantiomers were derivatized with a chiral agent (2S,4R)-N-heptafluorobutyryl-4-heptafluorobutoyloxy-prolyl chloride, developed at the authors laboratory and quantified by gas chromatography coupled to mass spectrometry operating in the negative chemical ionization mode. Most of the hair specimens that were tested positive for MDMA showed a predominance of the (R)-enantiomer. The R/S ratios of MDMA varied between 1.02 and 2.75 and total concentrations ranged from 0.1 to 20.1 ng/mg. The enantiomers of its metabolite 3,4-methylenedioxyamphetamine (MDA) were also quantified in most hair segments. The R/S ratios of MDA varied between 0.60 and 1.60, while the concentrations of the enantiomers ranged from 10 to 160 pg/mg hair. When segmental analysis was performed on single hair specimens, no inversion of the R versus S ratios of MDMA and MDA was observed. The predominance of (R)-MDMA in hair was in accordance with those already published for other matrices. Furthermore, both enantiomers of amphetamine (AM) were also detected in hair segments of four volunteers and the R/S ratios ranged from 1.00 to 1.47.

Trichopus zeylanicus combats fatigue without amphetamine-mimetic activity.

Chronic fatigue is a complex and little understood symptom for which there is no safe and effective pharmacotherapy. The present study was conducted to investigate the effectiveness of Trichopus zeylanicus whole plant powder on fatigue in young Sprague Dawley rats, and aged normal and long-living mutant Ames dwarf mice. Fatigue was evaluated by subjecting the animals to a forced swim test. Trichopus zeylanicus (250 and 500 mg/kg) treated young Sprague-Dawley rats resisted fatigue at a significant level (p < 0.005) compared with controls by an extended swim time in the forced swim test. Oral Trichopus zeylanicus (500 mg/kg) treatment for 2 weeks significantly increased the mobility time in the aged mutant (p < 0.05) and normal mice (p < 0.01) and significantly increased the swim time in the forced swim test in the aged normal mice (p < 0.05). Amphetamine-mimetic activity in Trichopus zeylanicus was excluded by suitable tests. These results show that Trichopus zeylanicus whole plant powder has anti-fatigue effects in young Sprague-Dawley rats and aged normal and mutant Ames dwarf mice providing scientific evidence for the Kani tribal practice in India.

Screening for amphetamine and amphetamine-type drugs in doping analysis by liquid chromatography/mass spectrometry.

A selective and sensitive method for the qualitative screening of urine samples for 27 amphetamine and amphetamine-type drugs in the field of doping analysis is described. The method consists of a liquid-liquid extraction with diethyl ether at pH 14 and analysis of the extracts with a LCQ-Deca mass spectrometer equipped with an atmospheric pressure chemical ionisation interface, operated in positive ionisation mode. The total run time was 15 min. All compounds were analysed in MS2 or MS3. The detection limit for all compounds was lower than 25 ng/mL except for chlorphentermine (detection limit: 250 ng/mL).

Simultaneous enantioselective determination of amphetamine and congeners in hair specimens by negative chemical ionization gas chromatography-mass spectrometry.

Enantioselective quantification of amphetamine (AM), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyethylamphetamine (MDEA) enantiomers in hair using gas chromatography-mass spectrometry (GC-MS) is described. Hair specimens were digested with 1M sodium hydroxide at 100 degrees C for 30 min and extracted by a solid phase procedure using Cleanscreen ZSDAU020. Extracted analytes were derivatised with (S)-heptafluorobutyrylprolyl chloride and the resulting diastereoisomers were quantified by GC-MS operating in the negative chemical ionization mode. Extraction yields were between 73.0 and 97.9%. Limits of detection varied in the range of 2.1-45.9 pg/mg hair, whereas the lowest limits of quantification varied between 4.3 and 91.8 pg/mg hair. Intra- and inter-assay precision and respective accuracy were acceptable. The enantiomeric ratios (R versus S) of AM, MA, MDA, MDMA and MDEA were determined in hair from suspected amphetamine abusers. Only MA and AM enantiomers were detectable in this collective and the quantification data showed in most cases higher concentrations of (R)-MA and (R)-AM than those of the corresponding (S)-enantiomers.

Gas chromatography-high-resolution mass spectrometric method for determination of methamphetamine and its major metabolite amphetamine in human hair.

Gas chromatography-high-resolution mass spectrometric (GC-HRMS) method is presented for the qualitative and quantitative analysis of methamphetamine (MA) and its major metabolite, amphetamine (AMP), in human hair. The method procedure involves decontamination of hair with distilled water and acetone, acidic hydrolysis and extraction in the presence of the internal standard, and GC-HRMS selective ion monitoring (SIM) analysis. The limits of detection (LOD) were 9 pg/mg for MA and 21 pg/mg for AMP using a 30-mg hair sample, and the SIM responses were linear with coefficients of correlation ranged from 0.9998 to 0.9999. The recoveries were found to be 91.1-92.3%. By using HRMS (resolution of 5000), detection sensitivity is improved because of the elimination of the biological background, and the LODs for MA and AMP were 2.4-4.4 times lower than those of low-resolution MS. The GC-HRMS method was successfully applied to the analysis of cosmetically treated hair, which is difficult to analyze with the conventional method.

Sensitive and simple gas chromatographic-mass spectrometric determination for amphetamine in microdialysate and ultrafiltrate samples.

A gas chromatographic-mass spectrometric (GC-MS) method is described for the measurement of amphetamine (AMP) using negative chemical ionization (NCI) mode. Without prior extraction AMP was derivatized with 2,3,4,5,6-pentafluorobenzoyl chloride (PFBC) and simultaneously extracted into toluene. The toluene extract was injected directly into GC-MS equipped with a HP-1 capillary column. The method is simple and more sensitive than most of the previously published methods. The limit of quantification of amphetamine is 25pg (1.4pg on column) with a very limited sample volume (25microl). The within-day precision was from 1.7 to 5.1% and between-day precision was from 2.2 to 7.3%. The method has been used for the measurement of several thousand microdialysate and ultrafiltrate samples and proven reliable.