Optimization of a simple method for the chiral separation of methamphetamine and related compounds in clandestine tablets and urine samples by beta-cyclodextrine modified capillary electrophoresis: a complementary method to GC-MS

The chiral separation of (+/-)-methamphetamine, (+/-)-methcathinone, (+/-)-ephedrine and (+/-)-pseudoephedrine by means of beta-cyclodextrine modified capillary electrophoresis is described. The distribution of enantiomers in clandestine tablets and urine samples were identified. Several electrophoretic parameters such as the concentration of beta-cyclodextrin, temperature, the applied voltage and the amount of organic solvent required for successful separation were optimized. The method, as described herein, represents a good complementary method to GC-MS for use in forensic and clinical analysis.     

Simultaneous separation of different types of amphetamine and piperazine designer drugs by capillary electrophoresis with a chiral selector

The recent emergence of a new class of piperazine-type compounds has brought about the need for laboratory screening methods for both seized drugs and toxicological samples. These piperazine compounds, which include 1-benzylpiperazine (BZP) and 1-(3-trifluoromethylphenyl)piperazine (TFMPP), exhibit comparable physiological effects and can be substituted for the classic amphetamine-type drugs. We have optimized a chiral capillary electrophoresis (CE) separation that detects a set of 6 piperazine and 4 chiral amphetamine compounds in under 23 min using a 200 mM phosphate buffer at a pH = 2.8 with 20 mM hydroxypropyl- beta-cyclodextrin (HPbeta3CD). In addition to the above compounds, a series of "clandestine" BZP diHCl samples were also analyzed using this method to assess the ruggedness of the procedure. The novel CE separation was tailored to simultaneously detect these piperzine compounds in addition to amphetamine-type drugs. Distinct migration time and UV-spectral data were obtained for all compounds of interest.     

Determination of Inorganic Ion Profiles of Illicit Drugs by Capillary Electrophoresis

A portable capillary electrophoresis instrument with dual capacitively coupled contactless conductivity detection (C⁴D) was used to determine the inorganic ionic profiles of three pharmaceutical samples and precursors of two illicit drugs (contemporary samples of methylone and para‐methoxymethamphetamine). The LODs ranged from 0.10 μmol/L to 1.25 μmol/L for the 10 selected cations, and from 0.13 μmol/L to 1.03 μmol/L for the eight selected anions. All separations were performed in less than 6 min with migration times and peak area RSD values ranging from 2 to 7%. The results demonstrate the potential of the analysis of inorganic ionic species to aid in the identification and/or differentiation of unknown tablets, and real samples found in illicit drug manufacture scenarios. From the resulting ionic fingerprint, the unknown tablets and samples can be further classified.     

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.     

Capillary electrophoresis: a new tool in forensic toxicology. Applications and prospects in hair analysis for illicit drugs

Capillary electrophoresis, the modern approach to instrumental electrophoresis, is probably the most rapidly expanding analytical technique that has appeared in recent years. In the hands of forensic toxicologists, capillary electrophoresis (CE) represents a powerful new analytical tool, which has proved suitable for the investigation of illicit drugs in seized preparations and also in complex biological matrices, among which is hair. CE can be applied according to different separation mechanisms, and among those that are toxicologically relevant are capillary zone electrophoresis and micellar electrokinetic capillary chromatography, which display different selectivities. For the investigation of hair for drugs of abuse, capillary electrophoresis proved effective, providing simultaneous determinations of different drugs without derivatization, with acceptable sensitivity (typically better than 1 ng of drug per mg of hair). The possibility of carrying out determinations of the same analytes, based on different separation mechanisms (capillary zone electrophoresis and micellar electrokinetic chromatography) with the same instrumentation, simply changing the buffer composition, provides an interesting possibility of ‘internal’ confirmation of the results.     

Study of TATP: stability of TATP solutions

Stability of raw TATP (3,3,6,6,9,9-hexamethyl-1,2,4,5,7,8-hexoxonane) samples in solutions of common solvents was studied to highlight problems faced by forensic labs in identification and analysis of organic peroxide samples. The TATP samples were prepared by reaction of acetone and hydrogen peroxide (30%) with the aid of following catalysts: hydrochloric, sulfuric, nitric, perchloric and methanesulfonic acid. Acetone, acetonitrile, methanol and acetonitrile/water solutions of TATP samples were prepared and stored at 50°C. Various degrees of stability were observed for particular combination of catalyst and solvent ranging from totally unstable (catalyst-H(2)SO(4)/any solvent) to very stable (catalyst-HCl/solvent acetonitrile). Purification of crude TATP by re-crystallization results in product stable in all investigated solvents. Stability of solution prepared from re-crystallized DADP (3,3,6,6-tetramethyl-1,2,4,5-tetroxane) was found to be on the same level as the stability of solution of re-crystallized TATP.     

Internal validation of reduced PCR reaction volume of the Qiagen Investigator® Argus X-12 QS Kit from blood samples on FTA cards

The onus of proof in criminal cases is beyond any reasonable doubt, and the issue on the lack of complete internal validation data can be manipulated when it comes to justifying the validity and reliability of the X-chromosomal short tandem repeats analysis for court representation. Therefore, this research evaluated the efficiency of the optimized 60% reduced volumes for polymerase chain reaction (PCR) amplification using the Qiagen Investigator® Argus X-12 QS Kit, as well as the capillary electrophoresis (CE) sample preparation for blood samples on Flinder's Technology Associates (FTA) cards. Good-quality DNA profile (3000–12,000 RFU) from the purified blood sample on FTA card (1.2 mm) were obtained using the optimized PCR (10.0 μL of PCR reaction volume and 21 cycles) and CE (9.0 μL Hi-Di™ Formamide and 0.3 μL DNA Size Standard 550 [BTO] and 27 s injection time) conditions. The analytical and stochastic thresholds were 100 and 200 RFU, respectively. Hence, the internal validation data supported the use of the optimized 60% reduced PCR amplification reaction volume of the Qiagen Investigator® Argus X-12 QS Kit as well as the CE sample preparation for producing reliable DNA profiles that comply with the quality assurance standards for forensic DNA testing laboratories, while optimizing the analytical cost.     

A quantitative densitometric determination of heroin and cocaine samples by high-performance thin-layer chromatography

The traditional thin-layer chromatography (TLC) is a sensitive technique frequently used in chemical analysis for semiquantitative determinations. In the last years a high-performance thin-layer chromatography system (HPTLC) has been developed. It assures quantitative determinations with direct densitometric measurements of chromatographic spots in situ on the plate. Here, we wish to report a method for the drug determination in street samples, that employs a CAMAG TLC/HPTLC Scanner Photodensitometer and a CAMAG LINOMAT III for sample application. Samples of illicit drug traffic (50 samples of street heroin and 30 samples of street cocaine) are tested and the gas-liquid chromatography (GLC) and HPTLC determinations are compared. The HPTLC method has proven to be sensitive, precise and accurate for quantitative determinations of drugs in street samples. Furthermore this method may be useful in forensic chemistry to show a "fingerprint" of each sample.     

Application of solvent microextraction to the analysis of amphetamines and phencyclidine in urine

A fast and simple method to detect some commonly abused illicit drugs, amphetamine, methamphetamine, 3,4-methylendioxy-amphetamine (MDA), 3,4-methylendioxy-methamphetamine (MDMA), 3,4-methylendioxy-N-ethylamphetamine (MDEA) and phencyclidine (PCP) in urine using solvent microextraction (SME) combined with gas chromatography (GC) analysis has been developed. The extraction is conducted by suspending a 2 microl drop of chloroform in a 2 ml urine sample. Following 8 min of extraction, the organic solvent is withdrawn into the syringe and injected into a GC with a pulsed discharge helium ionization detector (PDHID). The effects of different extraction solvents and times, pH and sample preparation were studied. The optimized method was capable of detecting drugs in urine at concentrations below Substance Abuse and Mental Health Services Administration (SAMHSA) established cut-off values for preliminary testing. Good linearity and reproducibility of extraction were obtained. The limits of detection were 0.5 microg/ml for amphetamine, 0.1 microg/ml for methamphetamine and MDA, 0.05 microg/ml for MDMA, 0.025 microg/ml for MDEA and 0.015 microg/ml for PCP. Relative standard deviation (R.S.D.) values ranged between 5 and 20% for the studied drugs.     

Developing a quantitative extraction technique for determining the organic additives in smokeless handgun powder.

Compositional analysis of the organic additives in smokeless handgun powder can provide forensic information to associate known and questioned samples. A reliable method for the quantitative extraction of smokeless powder additives would strengthen these measurements. To achieve quantitative recovery, both supercritical fluid extraction (SFE) and ultrasonic solvent extraction (USE) were evaluated as candidate techniques. Following a detailed evaluation of the solvent choice, the recovery of spiked additive compounds, and the effect of the powder matrix, a reliable USE technique was developed. When quantitative USE recovery of the target analytes, nitroglycerin (NG), diphenylamine (DPA), and ethyl centralite (EC), is coupled with additive measurement by micellar capillary electrophoresis (CE), compositional information can be obtained in less than 1 h.     

Occurrence of contamination by controlled substances in Euro banknotes from the Spanish archipelago of the Canary Islands

The social problems of drug abuse are a matter of increasing global problem. Nowadays, international agencies need fresh methods to monitor trends of the use of illicit drugs. In this sense, small amounts of drugs are transferred to banknotes and they could be detected and quantified. An analytical procedure based upon extraction with organic solvent, liquid chromatography separation, and mass spectrometric detection allowed the identification of 21 drugs and metabolites in 120 used Euro banknotes collected in the Canary Islands (Spain). Most of the banknotes analyzed showed detectable drug residues (92.5%). Cocaine was the most frequently detected drug, present in approximately 90% of the samples. In addition, 75%, 35%, and 15% of the banknotes showed residues of amphetamine derivatives, opiates, and benzodiazepines, respectively. An average of three drug residues per banknote was detected. In summary, the presence of drug residues in banknotes could be useful as tracer for drugs prevalence.     

Proton nuclear magnetic resonance spectroscopy (NMR) methods for determining the purity of reference drug standards and illicit forensic drug seizures

A rapid, sensitive, accurate, precise, reproducible, and versatile method for determining the purity of reference drug standards and the routine analysis of illicit drugs and adulterants using proton (1H) Nuclear Magnetic Resonance (NMR) Spectroscopy is presented. The methodology uses a weighed sample dissolved in a deuterated solvent or solvent mixture containing a high purity internal standard. The NMR experiment employs 8 scans using a 45 second delay and 90 degrees pulse. In the determination of purity of reference standards, the number of quantitative determinations available is equal to the number of peak groups that are baseline resolved. The relative standard deviation (RSD) of these signals is usually < 1% for pure standards, and the results agree well with other purity determining methods. This method can also aid in the determination of correct molecular weight for standards containing an unknown number of waters of hydration or an unknown number of acids per drug in salts. Because the molar response for the hydrogen nucleus is 1 for all compounds, and since no separation media are used, only one linearity study is required to test a probe. In the presented study, the linearity of the NMR probe was determined using methamphetamine HCl dissolved in deuterium oxide (D2O) with maleic acid as the internal standard (5 mg) for a range of concentrations from 0.033 to 69.18 mg/ml with a resulting correlation coefficient of >0.9999 for all 6 methamphetamine peak groups. The spectra of complex illicit heroin, methamphetamine, MDMA, and cocaine samples are presented, as well as an extensive list of compounds, their solubilities and the solvent(s) and internal standard used.     

Screening for the presence of drugs in serum and urine using different separation modes of capillary electrophoresis

Capillary electrophoresis (CE) is a modern separation technique that has some distinct advantages for toxicological analysis, such as a high efficiency, fast analysis, flexibility, and complementary separation mechanisms to chromatographic methods. CE can be applied in various modes, which each have a different separation mechanism or selectivity. The most common mode is capillary zone electrophoresis (CZE), in which charged analytes migrate in a buffer under the influence of an electric field. In micellar electrokinetic chromatography (MEKC), micelles are added to the buffer which interact with the analytes. MEKC can also be used for the separation of neutral compounds. In non-aqueous CE (NACE), the aqueous buffer is replaced by a background of electrolytes in organic solvents. A sample that needs to be screened can easily be analyzed subsequently by these CE modes using the same instrumentation.The aim of the study was to develop procedures for the analysis of basic and acidic drugs in serum and urine using CZE, MEKC, and NACE. A test mixture that consisted of six basic and six acidic compounds was used to study the separation behavior of five CE methods. The results showed that three methods (based on CZE, MEKC, and NACE) were suitable for the analysis of basic compounds and three methods (based on CZE and MEKC) for the analysis of acidic compounds.For the extraction of analytes from serum and urine, a solid-phase extraction (SPE) and a liquid-liquid extraction (LLE) method were compared. Both SPE and LLE methods provided clean extracts after extraction of the basic compounds from serum and urine. The extracts of acidic compounds contained more matrix interferences, especially for urine. The SPE method had some advantages compared to LLE, as it lead to cleaner extracts and higher peaks, and as it elutes basic and acidic compounds in one fraction.The potentials and pitfalls of the various methods for screening purposes in analytical toxicology are discussed.     

Application of solvent microextraction to the analysis of amphetamines and phencyclidine in urine

A fast and simple method to detect some commonly abused illicit drugs, amphetamine, methamphetamine, 3,4-methylendioxy-amphetamine (MDA), 3,4-methylendioxy-methamphetamine (MDMA), 3,4-methylendioxy-N-ethylamphetamine (MDEA) and phencyclidine (PCP) in urine using solvent microextraction (SME) combined with gas chromatography (GC) analysis has been developed. The extraction is conducted by suspending a 2 μl drop of chloroform in a 2 ml urine sample. Following 8 min of extraction, the organic solvent is withdrawn into the syringe and injected into a GC with a pulsed discharge helium ionization detector (PDHID).The effects of different extraction solvents and times, pH and sample preparation were studied. The optimized method was capable of detecting drugs in urine at concentrations below Substance Abuse and Mental Health Services Administration (SAMHSA) established cut-off values for preliminary testing. Good linearity and reproducibility of extraction were obtained. The limits of detection were 0.5 μg/ml for amphetamine, 0.1 μg/ml for methamphetamine and MDA, 0.05 μg/ml for MDMA, 0.025 μg/ml for MDEA and 0.015 μg/ml for PCP. Relative standard deviation (R.S.D.) values ranged between 5 and 20% for the studied drugs.     

Review of analytical methods for screening and quantification of fentanyl analogs and novel synthetic opioids in biological specimens

Fentanyl, fentanyl analogs, and other novel synthetic opioids (NSO), including nitazene analogs, prevail in forensic toxicology casework. Analytical methods for identifying these drugs in biological specimens need to be robust, sensitive, and specific. Isomers, new analogs, and slight differences in structural modifications necessitate the use of high-resolution mass spectrometry (HRMS), especially as a non-targeted screening method designed to detect newly emerging drugs. Traditional forensic toxicology workflows, such as immunoassay and gas chromatography mass spectrometry (GC–MS), are generally not sensitive enough for detection of NSOs due to observed low (sub-μg/L) concentrations. For this review, the authors tabulated, reviewed, and summarized analytical methods from 2010–2022 for screening and quantification of fentanyl analogs and other NSOs in biological specimens using a variety of different instruments and sample preparation approaches. Limits of detection or quantification for 105 methods were included and compared to published standards and guidelines for suggested scope and sensitivity in forensic toxicology casework. Methods were summarized by instrument for screening and quantitative methods for fentanyl analogs and for nitazenes and other NSO. Toxicological testing for fentanyl analogs and NSOs is increasingly and most commonly being conducted using a variety of liquid chromatography mass spectrometry (LC–MS)-based techniques. Most of the recent analytical methods reviewed exhibited limits of detection well below 1 μg/L to detect low concentrations of increasingly potent drugs. In addition, it was observed that most newly developed methods are now using smaller sample volumes which is achievable due to the sensitivity increase gained by new technology and new instrumentation.     

Sub-sampling and preparing forensic samples for pollen analysis

The main forensic application of palynology is in providing associative evidence, assisting to prove or disprove a link between people and objects with places or with other people. Although identification and interpretation of pollen is a specialist job, sub-sampling and preparing pollen samples for analysis may be carried out by non-specialists. As few forensic laboratories have residing palynologists, laboratories may wish to reduce the cost of analysis or risk of contamination by doing their own sub-sampling and preparation. Presented is a practical guide for sub-sampling and preparing forensic samples for pollen analysis, providing a complete standard procedure for both the palynologist and non-specialist. Procedures for sub-sampling include a wide variety of materials commonly collected for forensic analysis (soil, clothing and other fabrics, footwear, twine and rope, firearms, granulated materials, plant and animal material, and illicit drugs), many of which palynologists will not be familiar with. Procedures for preparation of samples (pollen concentration) are presented as a detailed, step-by-step method. Minimizing the risks of laboratory and cross-sample contamination during sub-sampling and preparation is emphasized.     

A review of impurity profiling and synthetic route of manufacture of methylamphetamine, 3,4-methylenedioxymethylamphetamine,amphetamine, dimethylamphetamine and p-methoxyamphetamine

Amphetamine-type substances (ATS), like other synthetically derived compounds, can be produced by a multitude of synthetic pathways using a variety of precursors and reagents, resulting in a large number of possible contaminants (by-products, intermediates and impurities). This review article describes the common contaminants found in preparations of methylamphetamine (MA), 3,4-methylenedioxymethylamphetamine (MDMA), amphetamine (AP), N,N-dimethylamphetamine (DMA) and p-methoxyamphetamine (PMA) synthesised via common synthetic pathways including reductive amination, Leuckart method, Nagai method, Emde method, Birch reduction, “Moscow” method, Wacker process, “Nitrostyrene” method and the Peracid oxidation method.Contaminants can facilitate identification of the synthetic route, origin of precursors and may suggest information as to the location of manufacture of these illicit drugs. Contaminant profiling can provide vital intelligence for investigations in which linking seizures or identifying the synthetic pathway is essential. This review article presents an accessible resource; a compilation of contaminants resulting from a variety of manufacturing methods used to synthesise the most common ATS. It is important for research in this field to continue as valuable information can be extracted from illicit drug samples, increasing discrimination amongst ATS, and in turn, leading to an increase in evidential value and forensic drug intelligence from forensic drug samples.     

GC-MS法测定血中1-甲基海因含量及其法医学应用

目的建立全血中1-甲基海因的气相色谱-质谱(GC-MS)分析方法,为1-甲基海因相关法医学鉴定案件提供技术支持。方法取0.5 m L血样,加入500 ng内标盐酸双苯戊二氨酯(SKF_(525A))、0.01 mol/L稀盐酸溶液2 m L,加入碳酸铵0.5 g调节p H值为9,加入乙酸乙酯2 m L,然后离心,取有机溶剂层,吹干后进行GC-MS分析。结果血液中1-甲基海因在0.5~50 ng/m L范围内线性关系良好,线性回归方程为y=0.015 51 x+0.007 26(R~2=0.999 7),最低检出限为0.1 ng/m L,回收率为93.02%~108.12%,日内、日间精密度分别小于6.07%和13.37%。结论本方法测定所得结果准确,重现性好,可用于血样中1-甲基海因含量的测定。     

Optimum methamphetamine profiling with sample preparation by solid-phase microextraction

Solid-phase microextraction (SPME) is a relatively new technique in which a small, polymer-coated fiber is employed to extract volatile and semivolatile organic compounds from the sealed headspace above a questioned sample. SPME, coupled with gas chromatography/mass spectrometry (GC/MS), was used to characterize impurities in illicit methamphetamine samples. Trace impurities present in a specimen were tentatively identified using mass-spectral databases and included 1,2-dimethyl-3-phenyl-aziridine (indicating synthesis via a halogenated ephedrine intermediate), ethyl vanillin (a flavoring compound), and caffeine (a stimulant used as cutting agent). The types and numbers of organic compounds sampled by SPME were compared with those collected by various solvent extraction protocols. In addition to unambiguously confirming the presence of methamphetamine, SPME-GC/MS analyses detected approximately 30 more organic analytes than were found by GC/MS following the ethyl acetate extraction method adopted by the United Nations International Drug Control Programme. SPME-GC/MS is a superior method for generating material "fingerprint" profiles in methamphetamine samples. The detection and characterization of increased points of comparison in drug samples provide more detailed chemical signatures for both intelligence and operational information.     

Solid-phase cation exchange extraction of basic drugs from the urine of racing greyhounds

The extraction of basic drugs from the urine of racing greyhounds has been carried out for many years using solvent extraction. This paper describes an extraction method that utilises a cation exchange column. Fourteen basic drugs were introduced onto the column in their ionised form, washed with organic and aqueous solvents and eluted with basic methanol. Three drugs, cyclizine, procaine and quinine were determined in authentic samples from racing greyhounds. A comparison of the solid-phase method with liquid-liquid extraction showed an increase in the efficiency of extraction and a reduction in both the time and cost incurred. The method requires a minimal volume of sample and solvent, little glassware and is easily automated.