Methamphetamine , 3,4-methylenedioxyamphetamine Research Articles

Fast on-site screening of 3,4-methylenedioxyethylamphetamine (MDEA) in forensic samples using carbon screen-printed electrode and square wave voltammetry

The worldwide seizures of illicit synthetic drugs have been dramatically increasing over the past decade, when the quantity of seized amphetamine-type stimulants (ATS) quadrupled, while seized ecstasy (3,4–methylenedioxymethamphetamine or MDMA)-like substances doubled. In this work, a simple and fast methodology for the electrochemical screening of ecstasy analogue MDEA (3,4-methylenedioxyethylamphetamine) in forensic samples is presented, for the first time, using a carbon screen-printed electrode (C-SPE) and square wave voltammetry (SWV). The electrochemical behavior of MDEA was studied by cyclic voltammetry at different pH values (2 to 12), where an altered electrochemistry of this drug on C-SPE, with an unreported redox pair, was observed in comparison with previous works dealing with the voltammetric detection of other ATS with similar chemical structures. Based on the acquired electrochemical processes, a redox reaction mechanism for MDEA on C-SPE is proposed. The best conditions (sensitivity and selectivity) for MDEA screening were obtained by SWV (a: 40 mV, f: 30 Hz, and Estep: 4 mV) and 0.04 mol L−1 Britton Robinson buffer solution at pH 3.0 as the supporting electrolyte. The proposed method presents a wide linear range (2.5 to 30.0 µmol L−1, R2 > 0.99), high sensitivity (0.569 µA / µmol L−1), low theoretical limit of detection (0.03 µmol L−1), and good repeatability using the same C-SPE (RSD < 6%; n = 10). The inter-device reproducibility (n = 5) showed a minimal variation of peak potentials (RSD < 2.3%). The interference study was performed with some adulterants usually found in ecstasy tablets, such as paracetamol, caffeine, glucose, ketamine, cocaine, acetylsalicylic acid, amphetamine (A), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), MDMA, ethylone (bk-MDEA), n-ethylpentylone (bk-EBDP) and dibutylone (bk-DMDBD). The method was applied in real seized samples containing MDEA with addition-recovery studies close to 100%. The developed method is promising to be used as a preliminary test for the screening of MDEA in the ever-changing situation concerning seized forensic samples. The proposed sensor exhibits attractive features for application in forensic analysis, such as rapidness, simplicity, low-cost, user friendly and portability.

What is the drug of choice of young festivalgoers?

BackgroundDrug and alcohol consumption are commonplace at festivals including those aimed at younger attendees. However, there is little quantitative information about the extent of this consumption. This work investigates drug use at a school-leaver festival and how it compares to non-festival weeks. MethodsInfluent wastewater was collected over three consecutive weeks from a location where a school-leaver festival occurs. Multiple liquid chromatography–mass spectrometry methods were used to analyse the use of illicit drugs, pharmaceuticals with abuse potential, new psychoactive substances (NPS), alcohol and cannabis. A method for human neurotransmitter metabolites was also utilised to show the population change and allow the drugs found to be normalised to a population. ResultsA total of 12 compounds were quantifiable: methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), alcohol, cannabis, cocaine, morphine, codeine, fentanyl, buprenorphine, oxycodone and nicotine. The NPS methylone was found solely over the festival weekend but at levels below the limit of quantification of the analytical method. The catecholamine metabolites vanillylmandelic acid (VMA) and homovanillic acid (HVA) were found over the entire three weeks, with identical trends – an increase over the festival weekend – indicating a population increase. HVA was used to normalise the drug mass loads to derive a population normalised mass load. Statistical differences using Hedges’ g showed large changes in the use of MDMA and MDA over the festival week. Smaller increases were also seen for alcohol and cocaine. ConclusionsThe drugs of choice for the attendees of this school-leaver festival were MDMA and MDA.

Gas chromatography-mass spectrometry (GC-MS) analysis of amphetamine, methamphetamine, 3,4-methylenedioxy-amphetamine and 3,4-methylenedioxymethamphetamine in human hair and hair sections

In this paper, the authors describe a sensitive method for simultaneous quantitation of amphetamine (AM), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA) in human hair by gas chromatography-mass spectrometry (GC-MS). Hair samples are initially cut into 0.5-cm sections, washed with methanol, and digested for 1 hr at 80°C in 2 N NaOH and deuterated internal standards. The resulting solutions are processed with a liquid-liquid extraction procedure and further derivatized with heptafluorobutyric anhydride (HFBA) before undergoing GC/MS analysis. The overall protocol achieves the following results when applied to the analysis of 50 mg drug-free hair specimens fortified with 2-40 ng/mg amphetamines: recovery: 77.45-86.86%; inter-day and intra-day precision ranges: 0.55-7.73% and 0.76-4.79%, respectively; linearity: r2 > 0.997; detection limits: 0.05 ng/mg for AM, MA, and MDMA and 0.1 ng/mg for MDA; and quantitation limits: 0.1 ng/mg for AM, MA, and MDMA and 0.2 ng/mg for MDA. Data derived from our analysis of hair samples collected from 30 self-reported methamphetamine abusers range from 1.39 to 15.41 (mean ± S.D., 5.21 ± 3.25) ng/mg for amphetamine and 12.58 to 173.28 (mean ± S.D., 56.10 ± 36.85) ng/mg for methamphetamine. This method has also been utilized successfully to evaluate the deposition pattern of drugs in head hairs collected from six female methamphetamine abusers staying at a rehabilitation facility. We conclude that this relatively simple protocol can be used for routine and reliable identification and quantitation of AM, MA, MDA, and MDMA in hair.

Rapid Analysis of Four Amphetamines in Urine by Self-Made Pipette-Tip Solid-Phase Extraction Followed by GC-MS/MS.

A simple and rapid pipette-tip solid-phase extraction (PT-SPE) procedure with derivatization prior to gas chromatography triple quadrupole mass spectrometry analysis is developed for the simultaneous determination of amphetamine (AMP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA) in urine samples. The PT-SPE procedure using self-made sorbent could extract drugs within 6min from 100-μL urine samples, requiring low solvent-consumption (<2.0mL). Besides, the self-made pipette tip could be reused at least five times. Under optimized conditions, the recoveries of four amphetamines at spiked levels (low, medium and high) ranged from 87.7 to 110.4%, with relative standard deviations<9.5%. The limit of detections and limit of quantifications for AMP, MA, MDA and MDMA were in the range of 2.52-8.25ng⋅mL-1 and 8.4-27.5ng⋅mL-1, respectively. Validation results show that the proposed method is suitable for the quantitation of amphetamines and has been successfully applied in the urine samples of suspected drug abusers.

SUPRAS extraction approach for matrix-independent determination of amphetamine-type stimulants by LC-MS/MS

Monitoring of amphetamine-type stimulant (ATS) confronts clinical labs with a high number of samples involving a variety of biological matrices. Liquid chromatography-tandem mass spectrometry (LC-MS/MS), routinely used for confirmation of ATS abuse, requires of laborious and matrix-dependent sample treatment methods, this increasing analysis time and cost. In this work, a universal and single-step sample treatment, based on supramolecular solvents (SUPRAS), was proposed for simplifying ATS confirmation in seven biological matrices. The SUPRAS was synthesized in situ in the sample (900 µL of basified oral fluid, urine, serum, sweat or breast milk or 50 mg of digested hair or fingernails) by the addition of hexanol (200 µL) and tetrahydrofuran (900 µL). The mixture was vortex-shaken and centrifuged and the SUPRAS extract was subsequently analyzed by positive ion mode electrospray LC-MS/MS. The method was fully validated for amphetamine (AMP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), N-ethyl-3,4-methylenedioxyamphetamine (MDEA) and N-methyl-3,4-methylenedioxyamphetamine (MDMA). Maximum ion suppression or enhancement was 9% and 7%, respectively, and extraction recoveries (87–111%) and within- (0.1–6.7%) and between-day (0.3–9.7%) CVs were all within required values. The lower limits of quantification (LLOQ) for biological fluids (5 ng/mL), and hair and fingernails (100 ng/g) were all well below the cut-offs established by worldwide organizations. Confirmation of MDA was carried out in five urine samples that tested positive for ATS by immunoassay. The SUPRAS-LC-MS/MS methodology succeeded in developing a hitherto unexplored and universal tool for quantifying ATS in a comprehensive pool of biological matrices of interest in forensic and clinical samples.

Novel Selectivity-Based Forensic Toxicological Validation of a Paper Spray Mass Spectrometry Method for the Quantitative Determination of Eight Amphetamines in Whole Blood.

Paper spray tandem mass spectrometry is used to identify and quantify eight individual amphetamines in whole blood in 1.3 min. The method has been optimized and fully validated according to forensic toxicology guidelines, for the quantification of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-methylamphetamine (MDMA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA), para-methoxyamphetamine (PMA), para-methoxymethamphetamine (PMMA), and 4-fluoroamphetamine (4-FA). Additionally, a new concept of intrinsic and application-based selectivity is discussed, featuring increased confidence in the power to discriminate the amphetamines from other chemically similar compounds when applying an ambient mass spectrometric method without chromatographic separation. Accuracy was within ±15% and average precision was better than 15%, and better than 20% at the LLOQ. Detection limits between 15 and 50 ng/mL were obtained using only 12 μL of whole blood. Graphical abstract ᅟ.

Statistical comparison of mass spectra for identification of amphetamine-type stimulants

A method for the statistical comparison of mass spectral data is demonstrated for applications in controlled substance analysis. The method uses an unequal variance t-test at each mass-to-charge ratio in the scan range to determine if two spectra are statistically associated or discriminated. If the two spectra are associated, a random-match probability is calculated to estimate the likelihood that the mass spectral fragmentation pattern in question occurs by random chance alone. If the two spectra are discriminated, the fragment ions responsible for the discrimination are determined.In this work, mass spectral data from case samples containing amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), phentermine, and psilocin were investigated. All spectra were collected in an accredited forensic laboratory using routine methods for controlled substance analysis. Using the statistical method, spectra of case samples were statistically associated to the corresponding reference standard at the 99.9% confidence level. In these instances, random-match probabilities ranged from 10−39 to 10−29, indicating the probability that the characteristic fragmentation pattern occurred by random chance is extremely small. Further, spectra of case samples were discriminated from other reference standards at the 99.9% or 99.0% confidence level, with 1–26 ions responsible for discrimination in each comparison.

Screening determination of four amphetamine-type drugs in street-grade illegal tablets and urine samples by portable capillary electrophoresis with contactless conductivity detection

A simple and inexpensive method for the identification of four substituted amphetamines, namely, 3,4-methylenedioxy methamphetamine (MDMA), methamphetamine (MA), 3,4-methylenedioxy amphetamine (MDA) and 3,4-methylenedioxy-N-ethylamphetamine (MDEA) was developed using an in-house constructed semi-automated portable capillary electrophoresis instrument (CE) with capacitively coupled contactless conductivity detection (C4D). Arginine 10mM adjusted to pH4.5 with acetic acid was found to be the optimal background electrolyte for the CE-C4D determination of these compounds. The best detection limits achieved with and without a sample preconcentration process were 10ppb and 500ppb, respectively. Substituted amphetamines were found in different seized illicit club drug tablets and urine samples collected from different suspected users. Good agreement between results from CE-C4D and those with the confirmation method (GC-MS) was achieved, with correlation coefficients for the two pairs of data of more than 0.99.

Quantification of illicit drugs in urine for forensic analysis in routine laboratories using ion trap LC-MSn

Aim Analysis of illicit drugs has become a major task in forensic and clinical research toxicology, doping control, workplace drug testing or law enforcement laboratories. Xenobiotics in biological samples are generally screened using various qualitative methods, often followed by targeted quantitative confirmation methods. In the past decades, liquid chromatography coupled to mass spectrometry has evolved to become the standard analytical technique for confirmatory analysis. Recently, a fast, robust and easy-to-use LC-MSn system was introduced for routine drug screening. In this study, quantitative analysis of amphetamines in extracted urine samples was carried out to demonstrate the applicability of the same instrument configuration for illicit drug confirmatory analysis, which enables usage of a single bench-top LC-MSn platform for high-confident multi-purpose analyses. Methods Authentic reference standard solutions of amphetamine (AMP), methamphetamine (MET), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA), methylbenzodioxolylbutanamine (MBDB) and their respective deuterated internal standards (IS) were used. The LC-MS analyses have been performed on an amaZon speed ion trap system with the ESI interface coupled to an UltiMate 3000 RSLC using an Acclaim RSLC 120 C18 column (2.2 μm; 2.1 × 100 mm) and a gradient system of acetonitrile/water/ammonium formate/formic acid as mobile phase. Data acquisition utilized SmartFrag and a novel smartMRM mode for high-sensitivity multi-target analysis. Data analysis combined library based unequivocal analyte identification and subsequent quantitation. Applicability of the method was tested with spiked urine matrix. The sample preparation was conducted by solid-phase extraction or by liquid-liquid extraction. Results The most common amphetamines and their respective internal standards were used to create a routine-ready quantitation method based on an actual LC-MSn screening platform. First, electrospray ionization conditions were optimized to achieve best selectivity and sensitivity for the target analytes. The chromatographic method was developed to efficiently separate the analytes while retaining the analysis time as short as possible. A novel targeted smartMRM mode enabled acquisition of full scan MS and MS2 spectra, and if necessary MS3, for multiple targeted experiments within defined retention time windows. Ideally, only the target analyte and its respective IS were present in a given time window: Minimal overlap of chromatographic peaks results in highest data acquisition rates for each target and thus yields highly reproducible and accurate values for a subsequent qualitative and quantitative analysis. Specificity, sensitivity, linearity, recovery and matrix effects were evaluated during method development with spiked urine samples extracted by LLE or SPE. Conclusion In contrast to LC triple quad instruments, MRM on an ion trap mass spectrometer provides full scan fragment information up to MSn which can be used for unambiguous identification and quantitation at the same time. In this way, primary analytical target identification was achieved by spectral library search based on full scan MSn spectra and retention time matching. Absolute quantitation, necessary to determine the actual analyte concentrations in a sample were achieved by generating MS2 extracted ion chromatograms of the main target ion transitions. According to the obtained results, the utilized LC ion trap system allows for qualitative library based screening but also enables quantitative target confirmation on the same instrument platform after thorough method development and validation.

Stereoisomeric profiling of drugs of abuse and pharmaceuticals in wastewaters of Valencia (Spain)

The enantiomeric and diastereomeric profiling of chiral pharmaceuticals (ephedrine, norephedrine, atenolol and venlafaxine) and illicit drugs (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-methylamphetamine (MDMA) and 3,4-methylenedioxy-N-ethylamphetamine (MDEA)) was undertaken over a period of fourteen consecutive days in three wastewater treatment plants (WWTPs) in the city of Valencia, Spain. Degradation efficiency of WWTPs was found to be compound and enantiomer dependent. Selective enantiomer enrichment was observed for several target analytes. Amphetamine and MDMA were enriched with R(−)-enantiomers. 1S,2S(+)-pseudoephedrine was found to be more readily degradable during activated sludge treatment than its diastereomer 1R,2S(−)-ephedrine. Atenolol underwent enrichment with either S(−)- or R(+)-enantiomer in different WWTPs. This unexpected enantiomeric variation in the stereoselective degradation of atenolol could be attributed to different processes utilized during activated sludge treatment. The application of (enantiomeric) profiling of wastewater revealed usage patterns of chiral drugs in the Valencia region.

Paper spray and extraction spray mass spectrometry for the direct and simultaneous quantification of eight drugs of abuse in whole blood.

Determination of eight drugs of abuse in blood has been performed using paper spray or extraction spray mass spectrometry in under 2 min with minimal sample preparation. A method has been optimized for quantification of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-methylamphetamine (MDMA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA), morphine, cocaine, and Δ9-tetrahydrocannabinol (THC) from a single blood spot. Sample to sample variations of 1-5% relative standard deviation were achieved using stable isotope-labeled internal standards and tandem mass spectrometry. Limits of detection for all drugs were below typical physiological and toxicological levels. Paper spray and extraction spray each used less than 10 μL of whole blood. These methods exhibit the potential for performing rapid and high-throughput assays for selective on-site multicompound quantitative screening of illicit drugs.

MALDI–TOF mass spectrometric determination of four amphetamines in blood

A rapid and sensitive detection method using matrix-assisted laser desorption ionization time-of-flight mass spectrometry was developed for the analysis of amphetamine (A), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethamphetamine (MDMA). In this method, α-cyano-4-hydroxy cinnamic acid was used as the matrix to assist the ionization of amphetamines. The MS spectra of these amphetamines showed protonated molecules [M + H]+ and fragment ions with comparable or higher intensities. The quantifications of A and MA were performed using A-d 7 and MA-d 5 as the internal standard, respectively, and those of MDA and MDMA were performed using MDMA-d 5 as the internal standard. The limit of detection and the quantification range using 20 μl of blood were about 10 ng/ml and 30–500 ng/ml for A and MDA, respectively, and about 1 ng/ml and 3–50 ng/ml for MA and MDMA, respectively. In two cases of poisoning in which MA was abused, the levels of A in blood and urine were from 0.050 to 3.24 μg/ml and the levels of MA were from 0.231 to 25.1 μg/ml.

Evaluation of amphetamine-type stimulant abuse through hair analysis: Results from 12 years of work.

Hair analysis is a reliable tool for detecting long-term exposure to illegal drugs, including amphetaminetype stimulants, over periods from a few weeks to a few months, depending on the length of the hair used for analysis. Between 2000 and 2012, over 600 hair samples were analysed at the Institute for Medical Research and Occupational Health, Croatia (IMROH) for the presence of amphetamine-type stimulants. IMROH has used the same procedure for testing hair samples for amphetamine-type stimulants for over twelve years. It was found to be reliable for confirming repeated abuse of amphetamine-type stimulants. Gas chromatography/mass spectrometry (GC/MS) was used to determine amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA-Ecstasy), and 3,4-methylenedioxyethylamphetamine (MDEA) in hair. Hair samples were either taken at the Institute, delivered by mail or a third person brought them to the laboratory. In most cases, the hair samples were tested anonymously. A total of 23 % of the tested samples were positive for one or more amphetamine-type stimulant. MDMA was the most frequently detected substance, whereas the most frequent combination was amphetamine with MDMA. Our results could indicate a trend in amphetamine-type stimulant abuse among young people in the Republic of Croatia.

Development of a method for the analysis of drugs of abuse in vitreous humor by capillary electrophoresis with diode array detection (CE–DAD)

This work presents the development of an analytical method based on capillary electrophoresis with diode array detection for the analysis of drugs of abuse and biotransformation products in vitreous humor. Composition of the background electrolyte, implementation of an online pre-concentration strategy and sample preparation procedures were objects of study. The complete electrophoretic separation of 12 analytes (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), ketamine, cocaine, cocaethylene, lidocaine, morphine, 6-monoacetylmorphine and heroin) and the internal standard N-methyl-1-(3,4-methylenedioxyphenyl)-2-butamine (MBDB) was obtained within 13min of run. The method was validated presenting good linearity (r(2)>0.99), recovery ≥90%, precision better than 12% RSD and acceptable accuracy in the range of 86-118% at three concentration levels (50, 100 and 500ng/mL). LODs and LOQs in the order of 1-5ng/mL and 5-10ng/mL, respectively, were obtained. After validation, the method was applied to eighty-seven vitreous humor samples and the results were compared to those obtained by a liquid chromatography tandem mass spectrometry (LC-MS/MS) screening method, routinely used by the forensic toxicology laboratory of the Sao Paulo State Police, Brazil. Cocaine was detected in 7.1%, cocaethylene in 3.6%, lidocaine in 2.4% and ketamine in 1.2% of the total number of analyzed samples.

Evaluation of pre‐analysis loss of dependent drugs in wastewater: stability and binding assessments

Wastewater analysis has the potential to provide objective and timely data on population drug consumption, but some crucial factors such as pre-analysis drug loss during sample storage and filtration could affect the accuracy and reliability of the method, and these uncertainties have yet to be fully assessed. This study was designed to evaluate analyte stability in wastewater stored under different conditions with the aim of optimizing the sample storage procedures for future studies. It also investigated whether there is significant analyte loss during filtration before sample extraction and storage after that. The studied substances and metabolites were: cotinine, cocaine and its metabolite benzoylecgonine, phenethylamines amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), opioids including codeine, methadone, 6-monoacetylmorphine (MAM) and morphine. In most situations, storing samples at 4 °C is sufficient to stabilize analytes for at least 2 weeks, and refrigeration is unnecessary during sample transportation within 3 days. However, additional measures need to be taken if unstable analytes such as cocaine and MAM are to be analyzed. No significant analyte loss was observed in the filtration process or in reconstituted extract stored at 4 °C or -20 °C for 2 weeks. By choosing stable analytes and proper storage conditions, wastewater analysis has the potential to provide accurate data for estimation of community drug use.

Comparison of Five Derivatizing Agents for the Determination of Amphetamine-Type Stimulants in Human Urine by Extractive Acylation and Gas Chromatography-Mass Spectrometry

Five acylation reagents have been compared for use as derivatizing agents for the analysis of amphetamine-type stimulants (ATS) in urine by gas chromatography-mass spectrometry (GC-MS). The evaluated reagents were heptafluorobutyric anhydride, pentafluoropropionic anhydride, trifluoroacetic anhydride, acetic anhydride (AA) and N-methyl-bis(trifluoroacetamide). The ATS included amphetamine, methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyethylamphetamine (MDEA). A mixture of the ATS was added to urine (1 mL) followed by KOH solution and saturated NaHCO(3) solution. The sample was then extracted with dichloromethane and the derivatizing agent and 2 µL were injected into the GC-MS instrument. The derivatizing agents were compared with reference to the signal-to-noise (S/N) ratios, peak area values, relative standard deviations (RSDs), linearities, limits of detection (LODs) and selectivities. The acetic anhydride proved to be the best according to the S/N ratio and peak area results for amphetamine, MA, MDMA and MDEA. The best RSD values of peak areas and of S/N ratios at 3 µg/mL were also given by AA in cases of MDA, MDMA and MDEA. At 20 µg/mL, the lowest RSD values of peak areas for MDA and the lowest RSD values of S/N ratios for MA, MDA, MDMA and MDEA were again given by AA. Additionally, the highest correlation coefficients for MA, MDA, MDMA and MDEA and the lowest LOD results for MA, MDMA and MDEA were produced by AA.

Estimation of gamma-hydroxybutyrate (GHB) co-consumption in serum samples of drivers positive for amphetamine or ecstasy

There is no toxicological analysis of gamma-hydroxybutyrate (GHB) applied routinely in cases of driving under influence (DUI); therefore the extent of consumption of this drug might be underestimated. Its consumption is described as occurring often concurrently with amphetamine or ecstasy. This study examines 196 serum samples which were collected by police during road side testing for GHB. The samples subject to this study have already been found to be positive for amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA) and/or 3,4-methylenedioxyethamphetamine (MDEA). Analysis has been performed by LC/MS/MS in the multiple reaction monitoring (MRM) mode. Due to its polarity, chromatographic separation of GHB was achieved by a HILIC column. To differentiate endogenous and exogenous levels of GHB, a cut-off concentration of 4μg/ml was applied. Of the 196 samples, two have been found to be positive for GHB. Of these samples, one sample was also positive for amphetamine and one for MDMA. Whilst other amphetamine derivates were not detected in these samples, both samples were found to be positive for cannabinoids. These results suggest that co-consumption of GHB with amphetamine or ecstasy is relatively low (1%) for the collective of this study.

New chlorinated amphetamine-type-stimulants disinfection-by-products formed during drinking water treatment

Previous studies have demonstrated high removal rates of amphetamine-type-stimulants (ATSs) through conventional drinking water treatments; however the behaviour of these compounds through disinfection steps and their transformation into disinfection-by-products (DBPs) is still unknown. In this work, for the first time, the reactivity of some ATSs such as amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyethylamphetamine (MDEA) with chlorine has been investigated under simulated and real drinking water treatment conditions in order to evaluate their ability to give rise to transformation products. Two new DBPs from these illicit drugs have been found. A common chlorinated-by-product (3-chlorobenzo)-1,3-dioxole, was identified for both MDA and MDEA while for MDMA, 3-chlorocatechol was found. The presence of these DBPs in water samples collected through drinking water treatment was studied in order to evaluate their formation under real conditions. Both compounds were generated through treatment from raw river water samples containing ATSs at concentration levels ranging from 1 to 15 ng/L for MDA and from 2.3 to 78 ng/L for MDMA. One of them, (3-chlorobenzo)-1,3-dioxole, found after the first chlorination step, was eliminated after ozone and GAC treatment while the MDMA DBP mainly generated after the postchlorination step, showed to be recalcitrant and it was found in final treated waters at concentrations ranging from 0.5 to 5.8 ng/L.

Electromembrane extraction of stimulating drugs from undiluted whole blood

For the first time, electromembrane extraction (EME) of six basic drugs of abuse from undiluted whole blood and post mortem blood in a totally stagnant system is reported. Cathinone, methamphetamine, 3,4-methylenedioxy-amphetamine (MDA), 3,4-methylenedioxy-methamphet-amine (MDMA), ketamine and 2,5-dimethoxy-4-iodoamphetamine (DOI) were extracted from the whole blood sample, through a supported liquid membrane (SLM) consisting of 1-ethyl-2-nitrobenzene (ENB) immobilized in the pores of a hollow fiber, and into an aqueous acceptor solution inside the lumen of the hollow fiber. The SLM acts as a barrier with efficient exclusion of all macromolecules and acidic substances in the sample. Due to the application of the electrical field, only the cationic compounds of interest are extracted efficiently across the membrane, thus providing extremely clean extracts for analysis with liquid chromatography–mass spectrometry, LC–MS. Recoveries in the range 10–30% were obtained from 80μl whole blood within 5min extraction time and an applied voltage of 15V across the SLM. The optimized technique was tested on real forensic whole blood samples taken from three forensic autopsy cases and on five forensic whole blood samples from living persons. The results were in agreement with the analysis using standard sample preparation methods (liquid–liquid extraction) performed on the same samples by Norwegian Institute of Public Health (NIPH), Division of Forensic Toxicology and Drug Abuse Research. Evaluation data were acceptable, with limit of detections (LODs) in the range 40–2610pg/mL, well below concentrations associated with drug abuse; linearites in the range between 10 and 250ng/mL with r2 values above 0.9939, and with repeatability (RSD) of 7–32%.

P.6.d.002 Activation of 5-HT1B receptors enhances the reinforcing activity of amphetamine in a self-administration model in rats

This work presents the development of an analytical method based on capillary electrophoresis with diode array detection for the analysis of drugs of abuse and biotransformation products in vitreous humor. Composition of the background electrolyte, implementation of an online pre-concentration strategy and sample preparation procedures were objects of study. The complete electrophoretic separation of 12 analytes (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), ketamine, cocaine, cocaethylene, lidocaine, morphine, 6-monoacetylmorphine and heroin) and the internal standard N-methyl-1-(3,4-methylenedioxyphenyl)-2-butamine (MBDB) was obtained within 13 min of run. The method was validated presenting good linearity (r2 > 0.99), recovery ≥90%, precision better than 12% RSD and acceptable accuracy in the range of 86–118% at three concentration levels (50, 100 and 500 ng/mL). LODs and LOQs in the order of 1–5 ng/mL and 5–10 ng/mL, respectively, were obtained. After validation, the method was applied to eighty-seven vitreous humor samples and the results were compared to those obtained by a liquid chromatography tandem mass spectrometry (LC-MS/MS) screening method, routinely used by the forensic toxicology laboratory of the Sao Paulo State Police, Brazil. Cocaine was detected in 7.1%, cocaethylene in 3.6%, lidocaine in 2.4% and ketamine in 1.2% of the total number of analyzed samples.