Separation and quantitation of optical isomers of methylamphetamine samples by capillary electrophoresis

A capillary electrophoretic (CE) procedure using an electrolyte modified with hydroxypropyl-β-cyclodextrin has been used for the separation and quantitation of (S)- and (R)-methylamphetamine in fifteen laboratory prepared samples and ten `street' samples. Excellent separation of (S)- and (R)-methylamphetamine was achieved and the levels of methylamphetamine in the samples were in good agreement with the levels determined by a separate CE procedure previously developed in our laboratory. Both CE systems showed the presence of impurities in some of the samples suggesting that CE could be used in profiling illicit methylamphetamine seizures. Gas chromatography-mass spectrometry (GC-MS) analysis of two of the samples identified one of the impurities as N-formyl methylamphetamine, however, no conclusive proof for the presence of this compound in the electropherograms was available. The chloride and sulphate content of the ten `street' samples was also determined by CE.     

Microbial degradation of illicit drugs,their precursors,and manufacturing by-products: implications for clandestine drug laboratory investigation and environmental assessment

Chemicals associated with clandestine drug laboratories are often disposed of covertly into soil, sewerage systems, or public waste management facilities. There are two significant issues relating to such dumps of materials; they might contain valuable evidence as to drug manufacture, and they might be a source of pollution. This study presents initial findings in relation to the impact microorganisms from environmental sources have upon drugs, their precursors, and manufacturing by-products. The aim of this study was to identify which chemicals associated with clandestine drug laboratories persist in the environment in order to allow forensic drug chemists to link discarded residues with the method of manufacture, and to allow the environmental impact of clandestine drug laboratories to be assessed accurately. When exposed to soil microorganisms, phenyl-2-propanone (P2P) was rapidly metabolized into mixtures of 1-phenyl-2-propanol, 1-phenyl-1,2-propanedione, 1-hydroxy-1-phenyl-2-propanone, 2-hydroxy-1-phenyl-1-propanone, and the two diastereoisomers of 1-phenyl-1,2-propanediol. On the other hand, when exposed under the same conditions, methylamphetamine sulphate (MAS) remained virtually unchanged. Implications relating to evidence gathering for forensic purposes and to environmental assessment of clandestine drug laboratories are discussed.     

Isolation and Identification of Three By‐products Found in Methylamphetamine Synthesized by the Emde Route

This article describes the isolation and structural elucidation of three compounds produced during the synthesis of methylamphetamine by the so‐called “Emde” procedure. The “Emde” procedure involves the preparation of the intermediate chloropseudoephedrine or chloroephedrine from ephedrine or pseudoephedrine, respectively. The intermediates are then reduced to methylamphetamine with hydrogen under pressure in the presence of a catalyst. The by‐product compounds were isolated from methylamphetamine by column chromatography and liquid chromatography (LC). Proton nuclear magnetic resonance spectroscopy (¹H NMR), carbon nuclear magnetic resonance spectroscopy (¹³C NMR), and nanospray quadrupole‐time of flight‐mass spectrometry (Q‐TOF‐MS) were used to identify them as two stereoisomers of the compound N, N′‐dimethyl‐3,4‐diphenylhexane‐2,5‐diamine and N‐methyl‐1‐{4‐[2‐(methylamino)propyl]phenyl}‐1‐phenylpropan‐2‐amine.     

Likelihood ratio methods for forensic comparison of evaporated gasoline residues

In the investigation of arson, evidence connecting a suspect to the fire scene may be obtained by comparing the composition of ignitable liquid residues found at the crime scene to ignitable liquids found in possession of the suspect. Interpreting the result of such a comparison is hampered by processes at the crime scene that result in evaporation, matrix interference, and microbial degradation of the ignitable liquid.Most commonly, gasoline is used as a fire accelerant in arson. In the current scientific literature on gasoline comparison, classification studies are reported for unevaporated and evaporated gasoline residues. In these studies the goal is to discriminate between samples of several sources of gasoline, based on a chemical analysis. While in classification studies the focus is on discrimination of gasolines, for forensic purposes a likelihood ratio approach is more relevant.In this work, a first step is made towards the ultimate goal of obtaining numerical values for the strength of evidence for the inference of identity of source in gasoline comparisons. Three likelihood ratio methods are presented for the comparison of evaporated gasoline residues (up to 75% weight loss under laboratory conditions). Two methods based on distance functions and one multivariate method were developed. The performance of the three methods is characterized by rates of misleading evidence, an analysis of the calibration and an information theoretical analysis.The three methods show strong improvement of discrimination as compared with a completely uninformative method. The two distance functions perform better than the multivariate method, in terms of discrimination and rates of misleading evidence.     

Underground pill testing, down under

At a recent South Australian rave, results reported to users from on-site pill-testing, using pill-testing kits, were compared with GCMS analysis of a scraping from the same pill. The presence of an ecstasy-like substance or methylamphetamine was correctly reported to users in 100% of pills that contained those substances. However only 11% of pills with combinations of illicit substances had both substances correctly identified. Ketamine was particularly problematic with identification occurring in only 18% of pills and in some instances, the colorimetric response obtained from ketamine was confused with the response from methylamphetamine. This study also allowed a comparison between pill design and composition encountered at the rave with those submitted to the forensic laboratory over a 6-month period including the month the rave was held. MDMA was present in 68% of pills at the rave and 89% of pills submitted by the police. Ketamine was present in 27 and 26% of pills, respectively and was often combined with other substances. The combinations of illicit substances were identical apart from one police-pill seizure that contained MDMA combined with PMA. This combination has not been previously encountered in South Australia. The pill designs observed at the rave differed significantly from the designs on pills submitted for testing by police. These differences limit the use of pill comparison charts as an alternative identification tool to colorimetric pill testing in South Australia.     

GC-MS and GC-IRD analysis of 2-, 3- and 4-methylmethamphetamine and 2-, 3- and 4-methylamphetamine

4-Methylmethamphetamine has been detected in samples submitted for analysis in several states throughout Australia. Six ring substituted methyl isomers of methamphetamine and amphetamine were synthesised and analysed. As the regioisomeric 2-, 3- and 4-methylmethamphetamine and 2-, 3- and 4-methylamphetamine have virtually identical mass spectra, the use of MS is an ineffective technique to discriminate between these closely related compounds. We set out to determine whether the regioisomers could be differentiated by a combination of GC-MS, acetyl derivatisation and GC-IRD. We demonstrate that the three isomers of methylmethamphetamine and methylamphetamine can be separated by GC, and a combination of acetyl derivatisation and vapour phase IR can identify the specific ring substituted compound.     

The Analysis of Black Powder Substitutes Containing Ascorbic Acid by Ion Chromatography/Mass Spectrometry*†

Abstract: Black powder substitutes containing ascorbic acid are a group of low explosives that utilize ascorbic acid as the fuel. The analysis of these powders is complicated by the degradation of ascorbic acid which occurs rapidly in solution and may also occur as the powder ages. Aqueous extracts of both intact powders and postblast residues were analyzed by an existing ion chromatography/mass spectrometry (IC/MS) method used at the Bureau of Alcohol, Tobacco, Firearms and Explosives. Results have shown that while ascorbic acid itself is not detected in this method, its diagnostic degradation products (threonic acid, monohydrated diketogulonic acid, and oxalic acid) can be identified. In addition, anions from the inorganic oxidizers (perchlorate and nitrate) and combustion products such as chloride, chlorate, and nitrite, can be identified within the same experiment. While this IC/MS method shows promise, future modifications are necessary because of limitations in identifying threonate in postblast residues, as well as coeluting compounds observed in postblast residues.     

N-cyanomethyl-N-methyl-1-(3',4'-methylenedioxyphenyl)-2-propylamine: an MDMA manufacturing by-product

This paper describes the structural elucidation of a compound produced during the synthesis of 3,4-methylenedioxymethylamphetamine (MDMA) via the reductive amination of 3,4-methylenedioxyphenyl-2-propanone (3,4-MDP-2-P) with methylamine and sodium cyanoborohydride. The compound was isolated from MDMA by column chromatography, proton and carbon nuclear magnetic resonance spectroscopy, LC/mass spectrometry, and total synthesis were used to identify the compound as N-cyanomethyl-N-methyl-1-(3',4'-methylenedioxyphenyl)-2-propylamine. This compound has been identified as a potential synthetic route marker for the reductive amination of 3,4-MDP-2-P with methylamine and sodium cyanoborohydride and as such it should prove valuable to forensic scientists engaged in profiling illicit drugs. Profiling MDMA can provide useful information to law enforcement agencies relating to synthetic route, precursor chemicals and reagents employed and may be used for comparative analyses of different drug seizures. This paper also describes the structural elucidation of the analogous methylamphetamine synthetic route marker compound, N-cyanomethyl-N-methyl-1-phenyl-2-propylamine, produced during the reductive amination of phenyl-2-propanone using methylamine and sodium cyanoborohydride.     

拉曼光谱快速定性分析合成毒品

目的快速定性检测合成毒品;方法利用激光拉曼光谱仪对合成毒品进行检验;结果合成毒品苯丙胺、甲基苯丙胺、氯胺酮、麻黄碱、甲基麻黄碱均能被拉曼准确检出;结论激光拉曼光谱能对合成毒品快速定性分析。     

The evaluation of the extent of transporting or "tracking" an identifiable ignitable liquid (gasoline) throughout fire scenes during the investigative process

Tests have determined that boots or shoes of individuals at a fire scene do not transport sufficient contaminants ("tracking") through the fire scene to produce a positive laboratory result for the presence of gasoline in a fire scene that was not present at the time of the fire. Questions about the validity of forensic laboratory results have been raised on the basis that low-level gasoline residues detected in the laboratory samples could have been the result of transporting the residue by footwear contaminated from the fire scene ("tracking"). The data collected in this study establish that "tracking" does not lead to false-positive laboratory results. Canines trained and experienced in the detection of trace ignitable liquid residues were also utilized in this study. The canine results confirmed that properly trained canines show a higher sensitivity than do standard ASTM laboratory techniques for fire debris analysis. In a few cases, canines responded to contamination, but laboratory testing (which is the definitive indicator) did not produce positive results.     

Comparing the effects of weathering and microbial degradation on gasoline using principal components analysis

Ignitable liquid residues recovered from a fire scene will often show signs of weathering as a result of exposure to the heat of the fire. In addition, when the substrate is rich in organic matter, both weathering and microbial degradation may be observed. In this study, 20 μL aliquots of fresh gasoline samples were intentionally weathered and also subjected to microbial degradation in potting soil. These samples were then analyzed using a passive adsorption-elution recovery method and gas chromatography/mass spectrometry. Peak areas from compounds of interest were normalized and autoscaled and then subjected to principal components analysis. This analysis showed that while lower boiling compounds are subject to weathering, a different set of compounds are subject to microbial degradation. Of the compounds studied, heptane, octane, toluene, and ethylbenzene were the most vulnerable to both weathering and microbial degradation. In contrast, 1,3,5-trimethylbenzene and 2-ethyltoluene were the most resistant to both phenomena.     

What is the best sample for determining the early postmortem period by on-the-spot flow cytometry analysis?

The level of degradation of DNA as a means for determining the time of death has been proposed as a valid adjunct to the classic thanatochronologic methods. The twofold aim of this work was to determine which organ might reveal both a correlation between the percentage of degradation of the DNA and the time lapse since death, and would be easiest to sample and yield the most reproducible results even in technically unfavorable situations such as on-the-spot investigations at the scene of death. A comparison of the spleen, blood, and liver showed that hepatic tissue best meets these specific needs because it shows a virtually linear correlation between the time elapsed since death and the level of degradation of the DNA, and it can easily be sampled at the scene of death by use of a common biopsy needle.     

Instability of calcium channel antagonists during sample preparation for LC-MS-MS analysis of serum samples

1,4-Dihydropyridines calcium channel antagonists (1,4-DHP CCAs) are photolabile and the products of their photodecomposition have no pharmaceutical activity. In our previous work we have presented a screening procedure for eleven 1,4-DHPs in plasma by LC-MS-MS using multiple reaction motoring. The laboratory process includes preparation and storage of stock solutions, plasma storage, solid-phase extraction, reconstitution of extracts and storage time in an autosampler for LC-MS-MS analysis. Prior to validation of the analytical procedure, we have tested the stability of these compounds by exposure to light. Methanolic solutions have been exposed to laboratory and UV light and the stability of the compounds in plasma was tested by exposure of spiked plasma samples to laboratory light at room temperature. Stability during freeze-thaw cycles and stability during 2 month storage at -20 degrees C have been tested as well. Products of photodecomposition have been identified after forced degradation and the degree of degradation has been quantified using LC-UV-DAD and LC-MS-MS, respectively. A 96% degradation after only 2h has been observed when solutions of nifedipine or nisoldipine were exposed to laboratory light in clear glass vials. In plasma samples degradation was 25% in only 2h for both compounds. The main degradation product was produced by oxidation of the dihydropyridinic ring resulting in the pyridine analogue that has been described as the first metabolite in the metabolic pathway. Only minor degradation was found for the other tested compounds after 2h light exposure in methanolic solutions. Furthermore, lercanidipine and nicardipine were also degradated by esterhydrolysis. Several additional minor degradation products were found for the other tested 1,4-DHPs, however, some of them could not be identified. Preconditions for storage and handling of plasma samples prior to and during analysis for 1,4-DHP CCAs are suggested in order to avoid photodecomposition of the analytes.     

UPLC-MS/MS法检测血痕中常见烟草、毒品和药物成分

目的建立UPLC-MS/MS测定血痕中尼古丁、可替宁、甲基苯丙胺、氯胺酮、吗啡、O6-单乙酰吗啡、地西泮、三唑仑、艾司唑仑、佐匹克隆和利血平的方法。方法以甲醇为提取液,采用基质提取溶液配制标准溶液制作定量曲线。采用超高效液相色谱柱对待测样品进行分离;以电喷雾离子源正离子（ESI＋）模式和多反应监测（MRM）模式进行质谱分析。优化实验条件,并进行方法学评价。结果选择UPLC HSS T3色谱柱,乙腈-5mmol/L甲酸铵＋0.1%（v/v）甲酸作为流动相,甲醇作为提取溶剂。11种检测物检出限（S/N=3）和定量限（S/N=10）分别为0.04~2.82ng和0.13~7.64ng,1~500ng/mL范围内的线性关系良好。除利血平、吗啡外的检测物回收率为（53.8±5.3）%~（107.2±12.6）%。结论采用本文UPLC-MS/MS方法对血痕中11种常见烟草、毒品和药物成分进行检测,能够满足实际检案的要求,可在相关检验中选用。     

Identification and quantification of p-hydroxyethylamphetamine as a novel metabolite of ethylamphetamine in rat by gas chromatography-mass spectrometry

Ethylamphetamine and two major metabolites in urine, obtained after intraperitoneal administration of the drug to rats, have been isolated by gas chromatography-mass spectrometry. We have also synthesized an authentic sample of p-hydroxy-N-ethylamphetamine by a simple method. In rat, the metabolic pathway of ethylamphetamine is similar to that of amphetamine and methylamphetamine according to quantitative analysis of the major metabolites using a selected ion monitoring (SIM) method.     

The potential of artificial aging for modelling of natural aging processes of ballpoint ink

Artificial aging has been used to reproduce natural aging processes in an accelerated pace. Questioned documents were exposed to light or high temperature in a well-defined manner in order to simulate an increased age. This may be used to study the aging processes or to date documents by reproducing their aging curve. Ink was studied especially because it is deposited on the paper when a document, such as a contract, is produced. Once on the paper, aging processes start through degradation of dyes, solvents drying and resins polymerisation. Modelling of dye's and solvent's aging was attempted. These processes, however, follow complex pathways, influenced by many factors which can be classified as three major groups: ink composition, paper type and storage conditions. The influence of these factors is such that different aging states can be obtained for an identical point in time. Storage conditions in particular are difficult to simulate, as they are dependent on environmental conditions (e.g. intensity and dose of light, temperature, air flow, humidity) and cannot be controlled in the natural aging of questioned documents. The problem therefore lies more in the variety of different conditions a questioned document might be exposed to during its natural aging, rather than in the simulation of such conditions in the laboratory. Nevertheless, a precise modelling of natural aging curves based on artificial aging curves is obtained when performed on the same paper and ink. A standard model for aging processes of ink on paper is therefore presented that is based on a fit of aging curves to a power law of solvent concentrations as a function of time. A mathematical transformation of artificial aging curves into modelled natural aging curves results in excellent overlap with data from real natural aging processes.     

A rapid and convenient LC/MS method for routine identification of methamphetamine/dimethylamphetamine and their metabolites in urine

A rapid and sensitive LC/MS method was developed for the simultaneous analysis of N,N-dimethylamphetamine (DMA), N,N-dimethylamphetamine N-oxide (DMANO), methylamphetamine (MA) and amphetamine (A) in urine samples. Employing an Alltech C18 column for solid phase extraction followed by LC/MS analysis using an Alltech Platinum EPS C18 column with a mixture of ammonium formate (0.01 M, pH 3) and acetonitrile (77:23, v/v) as mobile phase at a flow rate of 0.2 mL/min, simultaneous identification and quantitation of A, MA, DMA and DMANO in urine can be achieved using a 5-min chromatographic run. The calibration ranges were 0.10-3.0 micro g/mL for DMANO, 0.05-3.0 micro g/mL for DMA and 0.05-5.0 micro g/mL for both MA and A. The intra-, inter-day precision and accuracy for all analytes, spiked at three different concentrations in quality control samples, were in the ranges of 1.7-8.6, 4.1-10.0, -11.6 to 12.9%, respectively. The newly developed method was applied to the analysis of urine samples obtained from 118 suspected MA/DMA abusers, with the presence of MA confirmed in their urine samples under the drug-use surveillance program. Of these 118 samples, 43 were found to contain DMANO and 11 with both DMANO and DMA.     

Optimization and initial evaluation of 1,2-indandione as a reagent for fingerprint detection

1,2-Indandione has been used to develop fluorescent fingerprints on porous materials such as paper. The compound reacts with amino acid residues to produce highly fluorescent fingerprint ridges. An optimized formulation and treatment protocol for using the reagent is presented here. The reagent is applied as a solution in HFE7100 containing acetic acid and ethyl acetate. Treated articles are heated at 100 degrees C for 10 min at ambient humidity and stored in the dark before recording the fingerprints using fluorescence photography or digital imaging. Photodecomposition of the fluorescent fingerprints has been observed. Storage in the dark reduces degradation, extending the lifetime of the fingerprints. Other chemical methods to stabilize the fingerprints proved unsuccessful. Comparisons of the performance of 1,2-indandione with DFO in CFC113 performed on a limited range of substrates indicated that the reagent might be an effective method for the development of latent fingerprints despite the new reagent producing less intense fluorescence.     

A high-throughput urinalysis of abused drugs based on a SPE-LC-MS/MS method coupled with an in-house developed post-analysis data treatment system

A rapid urinalysis system based on SPE-LC-MS/MS with an in-house post-analysis data management system has been developed for the simultaneous identification and semi-quantitation of opiates (morphine, codeine), methadone, amphetamines (amphetamine, methylamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA)), 11-benzodiazepines or their metabolites and ketamine. The urine samples are subjected to automated solid phase extraction prior to analysis by LC-MS (Finnigan Surveyor LC connected to a Finnigan LCQ Advantage) fitted with an Alltech Rocket Platinum EPS C-18 column. With a single point calibration at the cut-off concentration for each analyte, simultaneous identification and semi-quantitation for the above mentioned drugs can be achieved in a 10 min run per urine sample. A computer macro-program package was developed to automatically retrieve appropriate data from the analytical data files, compare results with preset values (such as cut-off concentrations, MS matching scores) of each drug being analyzed and generate user-defined Excel reports to indicate all positive and negative results in batch-wise manner for ease of checking. The final analytical results are automatically copied into an Access database for report generation purposes. Through the use of automation in sample preparation, simultaneous identification and semi-quantitation by LC-MS/MS and a tailored made post-analysis data management system, this new urinalysis system significantly improves the quality of results, reduces the post-data treatment time, error due to data transfer and is suitable for high-throughput laboratory in batch-wise operation.