Amphetamine In Samples Research Articles

Nontarget screening of production waste samples from Leuckart amphetamine synthesis using liquid chromatography - high-resolution mass spectrometry as a complementary method to GC-MS impurity profiling.

The established approaches of suspect and nontarget screening (NTS) using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) are usually applied in the field of environmental and bioanalytical analysis. Herein, these approaches were employed on a forensic-toxicological application by analyzing different production waste samples from controlled amphetamine synthesis via Leuckart route to evaluate the suitability of this methodology for identification of route-specific organic substances in such waste samples. For analysis, two complementary LC techniques were used to cover a broad polarity spectrum. After data processing and peak picking using the enviMass software and further manual data restriction, 17 features were tentatively identified as suspects, three of which were subsequently identified with reference substances. All suspects had been previously identified in studies, in which gas chromatography-mass spectrometry (GC-MS) was successfully applied for synthesis marker assessment in waste and amphetamine samples. Remaining features with high signal intensity and assigned sum formula were selected for the attempt of structure elucidation. Seven potential synthesis markers were tentatively identified, which were not yet reported, except the sum formula of one compound, and which were partly also detected in real case waste samples afterward. The innovative application of the NTS approach using LC-HRMS for the analysis of aqueous amphetamine synthesis waste samples showed its suitability as extension to GC-MS analysis as it was possible to successfully identify seven new potential marker compounds, which are specific either for the conversion of the pre-precursors α-phenylacetoacetonitrile and α-phenylacetoacetamide to benzyl methyl ketone or for the subsequent Leuckart synthesis route after their conversion.

Identification of specific markers for amphetamines synthesized from glycidic acid pre-precursors and retrospective search in German profiling database.

The pre-precursor market and the clandestine production of amphetamine-type stimulants (ATS) has become more diverse in recent years. Besides α-phenylacetoacetonitrile (APAAN) and α-phenylacetoacetamide (APAA), glycidic acid derivatives and methyl α-phenylacetoacetate (MAPA) are gaining importance. This conclusion is based on seizure data of police and customs. However, analytical data are needed to confirm and quantify the actual prevalence of new pre-precursors by elucidating the percentage of seized ATS that have been produced from them. A recent study showed that APAAN use is currently declining, which supports the view that new pre-precursors are being used. In this study, several conversion procedures using different batches of glycidic acid derivatives and a complete Leuckart reaction to produce amphetamine were carried out. The resulting organic phases were analyzed using gas chromatography - mass spectrometry to identify possible marker compounds. Three marker compounds were discovered and characterized using mass spectra and nuclear magnetic resonance spectroscopy. They were identified as phenyl-1-propanone, N-(1-phenylpropyl)formamide and 1-phenylpropan-1-amine. Their prevalence was investigated by searching the markers in an amphetamine impurity profiling database to determine to what extent they occurred in amphetamine samples from recent years. Data from the central German amphetamine profiling database of more than 250 cases were used for this purpose. The yearly occurrence of the three glycidate marker compounds was determined going back as far as 2009, revealing an increasing trend from 2016 on. This article presents experimental proof that APAAN is currently being replaced by other pre-precursors, such as glycidic acid derivatives.

Enantiomeric profiling of amphetamine and methamphetamine in wastewater: A 7-year study in regional and urban Queensland, Australia

Enantiomeric profiling was used in this study to investigate the consumption of amphetamine and methamphetamine in regional and urban Southeast Queensland, Australia over a period of seven years. S(+) methamphetamine was predominantly consumed in both urban and regional areas, showing a two and three fold increase in urban and regional catchments respectively between 2011 and 2017. The ratio of amphetamine to methamphetamine (AMP/METH) in wastewater reflected the expected excretion profile of methamphetamine consumption indicating the presence of amphetamine in this study was primarily the result of methamphetamine metabolism. However, the occasional occurrence of R(−) amphetamine in samples containing higher AMP/METH ratios, suggested the consumption of racemic amphetamine. The R(−) methamphetamine enantiomer was also identified in several samples, possibly indicative that the phenyl-2-propanone (P2P) synthesis process rather than the more typical reduction of ephedrines was also being used to manufacture methamphetamine. Furthermore, we identified two samples with a significantly different enantiomer ratio for the METH and AMP as well as a much lower AMP/METH concentration ratio suggesting contribution from direct disposal of methamphetamine into the sewer. This study demonstrated that enantiomeric profiling in wastewater-based epidemiology can provide valuable information for evaluating the origin of amphetamine in wastewater as either a metabolite of methamphetamine consumption or amphetamine itself.

Identification of specific markers for amphetamine synthesised from the pre-precursor APAAN following the Leuckart route and retrospective search for APAAN markers in profiling databases from Germany and the Netherlands.

α-Phenylacetoacetonitrile (APAAN) is one of the most important pre-precursors for amphetamine production in recent years. This assumption is based on seizure data but there is little analytical data available showing how much amphetamine really originated from APAAN. In this study, several syntheses of amphetamine following the Leuckart route were performed starting from different organic compounds including APAAN. The organic phases were analysed using gas chromatography-mass spectrometry (GC-MS) to search for signals caused by possible APAAN markers. Three compounds were discovered, isolated, and based on the performed syntheses it was found that they are highly specific for the use of APAAN. Using mass spectra, high resolution MS and nuclear magnetic resonance (NMR) data the compounds were characterised and identified as 2-phenyl-2-butenenitrile, 3-amino-2-phenyl-2-butenenitrile, and 4-amino-6-methyl-5-phenylpyrimidine. To investigate their significance, they were searched in data from seized amphetamine samples to determine to what extent they were present in illicitly produced amphetamine. Data of more than 580 cases from amphetamine profiling databases in Germany and the Netherlands were used for this purpose. These databases allowed analysis of the yearly occurrence of the markers going back to 2009. The markers revealed a trend that was in agreement with seizure reports and reflected an increasing use of APAAN from 2010 on. This paper presents experimental proof that APAAN is indeed the most important pre-precursor of amphetamine in recent years. It also illustrates how important it is to look for new ways to identify current trends in drug production since such trends can change within a few years.

Impurity Profiling of Amphetamine and Methamphetamine Using Gas Chromatography Mass Spectrometry (GCMS) Harmonised Methods

Impurity profiling of drug seizures is a scientific approach employed to understand drug trafficking networks thus has becoming increasingly important in criminal investigation. This paper presents the feasibility of using the Collaborative Harmonisation of Methods for the Profiling of AmphetamineType Stimulants (CHAMP) established by the European Commission authority for impurity profiling of amphetamine and methamphetamine samples. Both drugs were analysed using similar extraction procedure and analytical conditions. The impurities were extracted from an alkaline buffer solution (pH8.1) using toluene prior to gas chromatography-mass spectrometry (GC-MS) analyses. The results showed that the reproducibility of the method for detecting amphetamine and methamphetamine ranged between 7.4-8.9 and 6.2-8.4 %RSD, respectively. Identification of impurities was performed by referencing against the available MS databases as well as to previous reported impurity profiling studies. Phenyl-2-propanone (P2P), also known as benzyl-methyl-ketone (BMK), as well as other specific impurities such as 4-methyl-5-phenylpyrimidine, bis-(1-phynelisopropyl) amine, N-formylamphetamine and N,N-di (b-phenylisopropyl) amines were identified in the amphetamine samples, indicating Leuckart’s pathway as the route of synthesis. Because P2P was also detected in the methamphetamine samples, the possible route of synthesis of the methamphetamine samples being Leuckart’s, nitrostyrene synthesis or reductive amination could not be ruled out.

Multivariate Curve Resolution-Alternating Least Squares Analysis of High-Resolution Liquid Chromatography-Mass Spectrometry Data.

Methods such as liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) are crucial for differentiating compounds with highly similar masses. This is a necessity when analyzing highly complex samples; however, the size of high-resolution LC-HRMS data sets can cause difficulties when applying advanced data analysis techniques. In this work, LC-HRMS analyses of known amphetamine samples and unknown bacterial lipid samples were carried out, and multivariate curve resolution-alternating least squares (MCR-ALS) was applied to the data to obtain mathematical separation of overlapped analyte signals. In order to minimize computational strain, a novel strategy was developed which minimizes the number of irrelevant masses analyzed at full resolution. To do this, data were first binned to unit mass resolution, and MCR-ALS was performed. This provided mathematical components for each analyte present plus background components. In the resolved spectral profiles of analyte components, masses above a preset intensity threshold were extracted, discarding all other masses, and expanded to successively higher levels of resolution, applying MCR-ALS at each level. These steps were repeated until 0.001 amu resolution was achieved, as dictated by the resolution of the instrument-in this case, a time-of-flight mass spectrometer. This strategy allowed for the accurate recovery of all known amphetamine compounds and select bacterial lipid extracts while minimizing the size of the data, therefore minimizing computational analysis time and data storage requirements. This relatively simple strategy enables the effective coupling of LC-HRMS with MCR-ALS.

A case story, involving the use of maltitol, a sugar alcohol, as a cutting agent in amphetamine and cocaine powders

Abstract In a criminal case involving cutting and resale of amphetamine and cocaine in the Copenhagen area of Denmark, maltitol was used as a cutting agent. The analysis of maltitol in seizures of pure diluents as well as in amphetamine and cocaine powders was carried out using reversed-phase high performance liquid chromatography (HPLC) with high-resolution (HR) mass spectrometric detection. Maltitol was identified in four out of nine amphetamine samples and in five out of six cocaine samples from the case in question. The use of maltitol as a cutting agent was considered by the police as a specific marker of the particular criminal group under investigation. To support or reject this hypothesis, cocaine and amphetamine samples from a four month period after the involved persons had been arrested were evaluated, also as part of the police investigation. None of these samples contained maltitol. The work described covers the part of the case involving the department of forensic chemistry, and not the whole police investigation, but everything was done within the frames given by the police. To the best of our knowledge, this is the first report of a disaccharide polyol being used as a cutting agent for illicit drugs.

Forensic analysis of P2P derived amphetamine synthesis impurities: identification and characterization of indene by-products.

1-Phenyl-2-propanone (P2P) is an internationally monitored precursor that has become increasingly difficult for illicit amphetamine producers to source, which means that alternative routes to its preparation have become increasingly important. One such approach includes the hydrolysis of alpha-phenylacetoacetonitrile (APAAN) with sulfuric acid. Previously, we reported the identification of 4,6-dimethyl-3,5-diphenylpryid-2-one following implementation of hydrolysis conditions and it was proposed that this compound might serve as one route specific by-product in the APAAN to P2P conversion. This study continued to explore the presence of impurities formed during this conversion and expanded also into a second route of P2P synthesis starting from alpha-methylstyrene (AMS). All P2P products underwent the Leuckart procedure to probe the presence of P2P-related impurities that might have carried through to the final product. Two by-products associated with the APAAN hydrolysis route to P2P were identified as 2,3-diacetyl-2,3-diphenylsuccinonitrile (1) and 2-methyl-1-phenyl-1,3-dicarbonitrile-1H-indene (2), respectively. Two by-products associated with the AMS route to P2P and subsequent Leuckart reaction were 1,1,3-trimethyl-3-phenyl-2,3-dihydro-1H-indene (3) and 1-phenyl-N-(phenylethyl)propan-2-amine (4), respectively. The two indenes (2 and 3) identified in synthesized amphetamine originating from P2P suggested that it might be possible to differentiate between the two synthetic routes regarding the use of APAAN and AMS. Furthermore, the association of these compounds with amphetamine production appears to have been reported for the first time. The presence of compounds 1 - 4 in seized amphetamine samples and waste products could facilitate the suggestion whether APAAN or AMS were employed in the synthesis route to the P2P. Copyright © 2016 John Wiley & Sons, Ltd.

Digital image‐based colourimetric tests for amphetamine and methylamphetamine

Digital image analysis was applied to the products of simple colorimetric [corrected] tests for amphetamine and methylamphetamine. Adobe Photoshop software was used for colour analysis to obtain analytical data in the form of a Red Green Blue (RGB) value. Calibration curves were developed for each compound and a number of illicit amphetamine samples were analyzed to demonstrate the application of the technique. A wide linear range and low detection limit for both drugs under test facilitated the application to illicit samples. Good agreement with gas chromatographic quantification results was obtained for the illicit samples analyzed by this method. The results show great potential for use as a semi-quantitative field test for illicit drug compounds.

Analysis of amphetamines illegally produced in Serbia

Forensic practice in the Republic of Serbia faced the illegal production of amphetamine for the first time in 2003. This paper presents the results of the chemical characterization of 32 batches of amphetamine samples from three separate cases, for the purpose of identification of the active components and additives. Through the profiling of impurities of all samples, using gas chromatography/mass spectrometry (GC/MS), 30 compounds associated with amphetamine were identified. The results of the analysis of powder tartrate, sulfate and phosphate salts of amphetamine, as well as variously formulated tablets are presented in this study. The analyses showed that the amphetamines were synthesized by the Leuckart method in all cases. .

Development of a harmonised method for the profiling of amphetamines VI: Evaluation of methods for comparison of amphetamine

Amphetamine samples were analysed by gas chromatography–mass spectrometry (GC–MS), and the peak areas of 33 target compounds were transformed by applying various pretreatment techniques. The objective was to optimise the ability of a number of distance metrics to establish links between samples of amphetamine originating from the same batch (henceforth refered to as linked distances). Furthermore, partial least squares discriminant analysis (PLS-DA) was used to evaluate the effects of various pretreatment methods on separation of amphetamine batches synthesised by the Leuckart reaction, reductive amination of benzyl methyl ketone, and the nitrostyrene route. The most efficient way to pretreat GC–MS data varied for the different distance metrics, although best results were obtained when data were normalised to the sum of peak areas, and either the fourth root or a logarithm was applied to the normalised data. When pretreating normalised data by fourth root transformation, Pearson correlation was the distance metric that was most successful at finding linked samples. Normalisation and the use of fourth root also represented the best method of pretreating data when employing PLS-DA to separate samples synthesised by different routes. To achieve a faster and more user-friendly procedure for evaluating chromatograms, experiments were performed in which the number of target compounds used to compare samples was reduced. The effect of each compound that was removed was studied by applying PLS-DA and by using Pearson correlation to calculate linked distances as well as unlinked distances (between samples from different batches of amphetamine). Considering both links between samples from the same batch and separation of samples synthesised by different routes, the best results were obtained with the data set comprising 26 compounds. Finally, it was found that the profiling method developed in this work was superior to an existing technique with respect to separating linked and unlinked distances.

Development of a harmonised method for the profiling of amphetamines: III. Development of the gas chromatographic method

This study focused on gas chromatographic analysis of target compounds found in illicit amphetamine synthesised by the Leuckart reaction, reductive amination of benzyl methyl ketone, and the nitrostyrene route. The analytical method was investigated and optimised with respect to introduction of amphetamine samples into the gas chromatograph and separation and detection of the target substances. Sample introduction using split and splitless injection was tested at different injector temperatures, and their ability to transfer the target compounds to the GC column was evaluated using cold on column injection as a reference. Taking the results from both techniques into consideration a temperature of 250 °C was considered to be the best compromise. The most efficient separation was achieved with a DB-35MS capillary column (35% diphenyl 65% dimethyl silicone; 30 m × 0.25 mm, d f 0.25 μm) and an oven temperature program that started at 90 °C (1 min) and was increased by 8 °C/min to 300 °C (10 min). Reproducibility, repeatability, linearity, and limits of determination for the flame ionisation detector (FID), nitrogen phosphorous detector (NPD), and mass spectrometry (MS) in scan mode and selected ion monitoring (SIM) mode were evaluated. In addition, selectivity was studied applying FID and MS in both scan and SIM mode. It was found that reproducibility, repeatability, and limits of determination were similar for FID, NPD, and MS in scan mode. Moreover, the linearity was better when applying FID or NPD whereas the selectivity was better when utilising the MS. Finally, the introduction of target compounds to the GC column when applying injection volumes of 0.2 μl, 1 μl, 2 μl, and 4 μl with splitless injection respectively 1 μl with split injection (split ratio, 1:40) were compared. It was demonstrated that splitless injections of 1 μl, 2 μl, and 4 μl could be employed in the developed method, while split injection and splitless injections of 0.2 μl should be avoided.

Comparison of microemulsion electrokinetic chromatography and solvent modified micellar electrokinetic chromatography on rapid separation of heroin, amphetamine and their basic impurities

Microemulsion electrokinetic chromatography (MEEKC) and solvent modified micellar electrokinetic chromatography (MEKC) were investigated with the goal of the rapid separation of complex heroin and amphetamine samples. The rapid simultaneous separation of 17 species of heroin, amphetamine and their basic impurities and adulterants was performed within about 10 min using MEEKC for the first time, whereas solvent modified MEKCs were unable to resolve all the components. The comparisons between MEEKC and solvent modified MEKC proved internal lipophilic organic phase in microemulsions played an important role in improving the separation performance with respect to efficiency. However, the role of internal lipophilic organic phase in MEEKC was disgusted at high concentrations of cosurfactant, and the separations of MEEKC and 1-butanol modified MEKC became similar at high concentrations of 1-butanol. The evaluation of reproducibility, linearity and detection limit of optimized MEEKC method provided good results for all the analytes investigated, thus allowing its application to real controlled drug preparation analysis.

Ecstasy (MDMA), Methamphetamine, and Date Rape (Drug-Facilitated Sexual Assault): A Consideration of the Issues

The term “date rape drug” has traditionally been applied by the media to powerful sedatives, such as gamma hydroxybutyrate (GHB) and flunitrazepam (Rohypnol), which can render a person unconscious and hence unable to resist and/or recall an assault. However, some law enforcement agents and others have recently obtained convictions by arguing that the empathy-generating and sensual effects of MDMA. and an occasional increase in disinhibition and sexual desire linked with methamphetamine use, remove a person's ability to give a reasoned consent, turning the person into “a helpless slave” to their own sexual desires and those of the alleged perpetrator. The argument holds that the victim becomes part of the assault because they may appear to be cooperating and colluding with activity which they would not have consented to without taking these drugs. This interpretation of the term “date rape” has been fed by data that sometimes finds MDMA and amphetamines in samples taken from sexual assault victims, and hence these prosecutions sometimes rely on expert testimony from toxicologists, pathologists and police officers rather than psychologists and psychiatrists who are expert in the human effects of these drugs. Some of those in the latter group have dismissed claims that MDMA is an aphrodisiac or a date rape drug as myths propagated by the media. In this article, these arguments and their respective strengths and weaknesses will be examined to assist professionals and others who may become involved in these cases.

Application of chemometric methods in searching for illicit Leuckart amphetamine sources

A chromatogram of contaminants of illicit amphetamine is referred to as contamination profile. The profile depends on the method and conditions of the drug synthesis. Therefore, it allows to link a drug sample sized by the police to the source of illicit production as well as enables to find out the links between dealers and users. In our study, statistical and chemometric methods were applied. The areas of 15 more informative chromatographic peaks were selected and used as original variables describing the drug samples. The selected peaks formed a profile. As a rule, samples submitted to the police laboratory are classified using correlation coefficient between the corresponding profiles as a measure of similarity. A preliminary classification and the affiliation of a given profile to a class is verified by means of principal component analysis and cluster analysis. Different distances between profiles are used including Euclidean and Pearson ones. The data structure is visualized in two- or three-dimensional spaces of two or three most important principal components. For the purpose of this study, a total of nearly 1000 drug samples were tested. It has been concluded that amphetamine samples may be attributed to one source if the Euclidean distance between the corresponding profiles is less than 1, and the Pearson distance is less than 0.04. This approach proved very effective in identification of drug traffic routes and clandestine laboratories by the police.

Determination of the enantiomeric purity of amphetamine after derivatization with Sanger's reagent (2,4-dinitrofluorobenzene [DNFB]) by simultaneous dual circular dichroism and ultraviolet spectroscopy

A new analytical procedure has been developed for the determination of the enantiomeric purity of amphetamine samples. It is based on the derivatization of amphetamine with 2,4-dinitrofluorobenzene and a subsequent measurement of the anisotropy factor by dual (CD/UV) spectroscopy. This factor depends on the enantiomeric composition and does not depend on the concentration, a feature that eliminates the systematic errors caused by the sample preparations such as dilution, weighing, etc. The main advantage of the approach is the fact that the new synthetized derivative can be measured in the visible wavelength region with significantly better sensitivity than the underivatized amphetamine. The applicability of the procedure is demonstrated by results obtained by analysis of model materials containing amphetamine enantiomers at different ratios.

Amphetamine profiling — improvements of data processing

A method to classify amphetamine samples according to their impurity content by the help of computer programs is described. In an example with made up data the method is shown and in a second example, with data from 136 amphetamine samples analysed during a short period, the agreement between this method and an independent visual classification of impurity profiles is good. The impurities are extracted by n-octane from an aqueous solution of amphetamine at pH 7 and analysed by gas chromatography. The integrated peak areas are used for classification in computer programs and examples of outputs are shown. By calculating principal components and displaying the data according to them a plot is shown on the computer screen where a classification may be found. By using a rotating plot more of the information can be used.

Street Drugs in Denmark

Abstract Street samples of heroin (n = 102) and amphetamine (n = 120) seized in different areas of Denmark during a one-year period were analyzed for purity and additive content. The mean concentrations of the heroin and the amphetamine samples were 34 and 35%, respectively, but the purity of both drugs varied greatly. Sugars and caffeine were the most frequently encountered cutting agents in both drugs. The additives phenobarbital, methaqualone, and procaine were seen only in heroin, especially in heroin base. Half of the heroin samples were heroin base, the other half heroin chloride. Brownish samples of heroin base containing large amounts of opium alkaloids dominated in the seizures in the provincial towns, whereas white/beige samples containing heroin chloride with little or no papaverine or noscapine were frequently seen among the seizures in the capital. The result may indicate different import routes for heroin to the eastern and western parts of Denmark.

Drug abuse in Denmark (Jutland and Funen). A forensic study based upon drugs seized in 1982-1987.

Two thousand fifty-three samples submitted to forensic chemical analysis during the period 1982-1987 were characterized according to their content of narcotics and the year of seizure. Heroin and cannabis dominated the material, but a significant decrease in the number of heroin samples was observed in 1987. Amphetamine, on the contrary, hardly seen during the period 1982-1984, accounted for half of the narcotics submitted in 1987. The misuse of morphine, especially in the form of poppy capsules, was a problem for some years, whereas cocaine has not yet been a drug of abuse in Denmark. The mean concentrations of the heroin and the amphetamine samples were 30% and 44%, respectively, both for retail drugs and the entire material.

Profiling of impurities in illicit amphetamine samples by high-performance liquid chromatography using column switching

A simple high-performance liquid chromatographic method, suitable for routine profiling of impurities in illegally produced amphetamine, has been developed. Amphetamine is dissolved in acetonitrile—citrate buffer (pH 3) (2:8) and injected directly without further sample pre-treatment. The impurities are enriched on-line on a C 8 extraction column, while amphetamine and polar diluents are washed out with water. After washing for 1.5 min, a six-port valve is switched and an acetonitrile-0.2 M butylamine in water (pH 8) gradient elutes the impurities from the extraction column on to a C 12 analytical column where they are separated. The compounds are monitored by UV detection at 220 and 254 nm. The total extraction and analysis time is 30 min. The method allows automated extraction and analysis to be performed.