Forensic Toxicology最新文献

Purpose: Etomidate, which is a psychoactive drug with an anesthetic effect, is used as a substitute for expensive mainstream drugs. There has been a trend toward abuse of etomidate and its emerging structural analogues now. Faced with a large number of samples, a rapid and effective detection method is needed.

Methods: In this study, ambient flame ionization (AFI) coupled with LTQ-Orbitrap mass spectrometry was used to analyze etomidate and its structural analogues in urine. It can realize detection in less than 0.2 min without sample preparation.

Results: Ideal analysis conditions were obtained by optimizing various parameters and analytical performance was validated. The isomers (isopropoxate and propoxate) can be distinguished by ion abundance ratios. Positive samples (n = 75) were analyzed very efficiently and successfully from plenty of authentic specimens (n = 116). Statistical analysis was conducted on drug types, age, and gender of drug users. A new structural analogue was discovered in one of the samples, which was a very crucial discovery. That meant the market may face with the emergence of new structural analogues.

Conclusions: This study can satisfy both targeted and non-targeted screening, which provides support for timely monitoring and detection of novel drugs and offers a wider range of method choices for forensic laboratories. It can also better cope with the current situation of drug control and combat crimes related to new types of drugs.

Purpose: Recently, a number of abuse and intoxication cases involving imidazole-derived GABA agonists etomidate and its analogs have been reported in China. During our recent screening of e-cigarette liquids, we encountered two novel etomidate analogs that have not previously been reported. This study aims to present the analytical procedures used to identify these compounds, along with detailed data obtained under various instrumental conditions.

Methods: Identification of the substances of concern was carried out using various instruments, including gas chromatography-mass spectrometry (GC-MS), headspace gas chromatography (HS-GC), liquid chromatography-high-resolution mass spectrometry (LC-HRMS), and liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Results: Analysis of the e-cigarette samples of concern using GC-MS, LC-HRMS, and LC-MS/MS yielded mass spectra, product ion mass spectra, and high-resolution mass spectra, which allowed structural elucidation. HS-GC analysis of hydrolyzed alcohols provided further insights into the substituent groups linked to imidazole core. Finally, the compounds were identified as butomidate and trifluoro-etomidate (tf-etomidate) through comparison with reference standards. Product ion mass spectra of the reference standards at various collision energies were obtained by LC-MS/MS, which also matched those of the target compounds.

Conclusion: This study demonstrated the ion spectra of butomidate, sec-butomidate and tf-etomidate as obtained by GC-MS, and their product ion spectra under varying collision energies by LC-MS/MS. These findings enabled the identification of the compounds in the dubious e-cigarette liquids. To our knowledge, this is the first report detailing qualitative procedures and associated instrumental data for the identification of butomidate and tf-etomidate.

Purpose: Δ⁹-Tetrahydrocannabinolic acid-A (Δ⁹-THCA-A) is a precursor of Δ⁹-tetrahydrocannabinol in cannabis. Here, considering applicability to ordinary forensic laboratories, we developed a novel isolation method for Δ⁹-THCA-A without the need for special equipment.

Methods: Dried pulverized cannabis inflorescence (2 g) was extracted with acetonitrile. After the extract was treated with graphite carbon powder to remove chlorophyll, the solvent was replaced with methanol. The methanol solution was diluted with aqueous sodium hydroxide solution and then washed with a mixture of n-hexane/ethyl acetate (7:1, v/v). The remaining aqueous layer was acidified with acetic acid and extracted with the n-hexane/ethyl acetate mixture. The extract was purified by silver nitrate-impregnated silica gel column chromatography.

Results: The isolation procedure gave a pale beige solid as the final product. The final product was identified as Δ⁹-THCA-A by comparison of the analytical results of gas chromatography/mass spectrometry (GC/MS) after trimethylsilylation and high-performance liquid chromatography with ultraviolet detection (HPLC-UV) with an authentic standard. The final product was confirmed to be highly pure by ¹H-nuclear magnetic resonance spectroscopy in addition to GC/MS and HPLC-UV.

Conclusions: This novel isolation method gave highly pure Δ⁹-THCA-A without using special purification equipment, such as a flash chromatography system or an HPLC system with a fraction collector, which are uncommon in forensic laboratories. This method will be useful for many forensic laboratories that find it difficult to obtain commercial Δ⁹-THCA-A as a standard.

Purpose: A new lysergic acid diethylamide (LSD) analog has recently been identified, 1-[3-(Trimethylsilyl)propanoyl] LSD (1S-LSD), characterized by a silicon-containing acyl moiety. In the proof of LSD analog consumption, direct detection of the parent compound in urine or blood can be challenging; therefore, characteristic metabolites as consumption markers should be detected. However, the metabolic fate is unclear. This study aimed to characterize the metabolic properties of 1S-LSD.

Methods: The synthesized 1S-LSD was incubated with human liver microsomes. The obtained metabolites were analyzed using liquid chromatography-quadrupole time-of-flight mass spectrometry.

Results: The parent compound was metabolized at a moderately rapid rate, with the early formation of LSD. Sixty-two metabolites were observed, and a metabolic pathway was proposed. The major metabolites were compounds with hydroxyl groups in the 3-silylpropanoyl moiety. Five metabolites were relatively abundant and retained their 3-silylpropanoyl moieties: N-deethylated 1S-LSD (Si04), N-deethylated and silanolized 1S-LSD (Si06), N-deethylated and monohydroxylated 1S-LSD (Si09 and Si11), and silanolized 1S-LSD (Si21).

Conclusions: The metabolic fate of 1S-LSD, an abused drug containing silicon, was characterized for the first time. The diverse metabolic pathways will help better understand the metabolic processes of not only 1S-LSD but also N¹-acylated LSD analogs and compounds with trimethylsilyl groups. Si04, Si06, Si09, Si11, and Si21 are potential target analytes for proving 1S-LSD consumption.

Purpose: Ethylene glycol (EG) is a typical antifreeze compound and a significant analyte in forensic toxicology. Current EG analytical method for biological samples in forensic toxicology employ liquid chromatography-tandem mass spectrometry (LC-MS/MS), however, they exhibit low sensitivity and reliability. Therefore, in this study, we aimed to establish a highly sensitive, selective, and reliable EG assay system for human serum analysis using a hydroxyl derivatization-aided LC-MS/MS technique.

Methods: p-Toluenesulfonyl isocyanate (PTSI) was applied for precolumn derivatization of EG in human serum, to enhance the sensitivity of LC-MS/MS for EG detection.

Results: The optimal derivatization conditions were 200 µL/mL PTSI in acetonitrile at 25 °C for 10 min. A highly sensitive and reliable LC-MS/MS detection of EG in human serum was achieved, with the calibration curve exhibiting a good linearity (r > 0.999, 10-1000 µg/mL of EG). The proposed PTSI-derivatization-LC-MS/MS method exhibited high reliability (1.4-1.8%) for the intra-day and inter-day repeatability (%RSD), and accuracy (96.7-102.4%), with the limits of detection and quantification in human serum being 0.023 µg/mL (S/N = 3) and 0.077 µg/mL (S/N = 10), respectively.

Conclusions: A novel PTSI derivatization-aided LC-MS/MS method was developed, offering a highly sensitive, selective, and reliable analytical tool for EG quantification in human serum for forensic toxicology applications.

Purpose: Aconitine is a highly toxic alkaloid found in Aconitum species, known for their potent neurotoxic and cardiotoxic effects. While accidental poisonings are relatively rare in Europe, intentional ingestions are more frequently reported. Despite the well-documented clinical effects of aconitine, a comprehensive toxicological investigation including analysis of hair and roots responsible for poisoning has never been reported.

Methods: A fatal case of aconitine poisoning was investigated following the ingestion of Aconitum roots. Biological samples (including hair) were analyzed using liquid chromatography-tandem mass spectrometry and liquid chromatography-high-resolution mass spectometry (LC-HRMS). The roots found at the victim's residence were also examined.

Results: Aconitine was detected in all tested biological matrices with concentrations of femoral blood and hair of 28.6 ng/mL and 54 pg/mg respectively. The amount of aconitine in the plant root was 0.6 mg/g. Based on the weight and number of roots ingested (as reported by the victim), the estimated dose of aconitine was 12 mg, approximately 2 to 4 times the known lethal dose for an adult.

Conclusion: This case presents the first detailed toxicological study of fatal aconitine poisoning that includes both hair and root analysis via LC-HRMS. The results highlight the value of advanced mass spectrometry in forensic detection of alkaloid exposure, while the development of a method for the identification of aconitine in hair could be useful in the future in reconstructing poisoning scenarios and assessing possible repeated exposures.

Purpose: Over-the-counter medicines are commonly used for recreational and suicidal overdoses, a global problem. Some of these are easily obtained via the Internet. In cases of intoxication, drug quantification is necessary to estimate the cause of death. Stable isotope compounds are recommended as internal standards (IS) for analyzing drugs; however, it is difficult for individual laboratories to obtain isotopes for all analytes due to cost and availability. Therefore, alternative IS selection is important for practicality. Here, we quantified diphenhydramine and dextromethorphan concentrations in plasma from several collection sites in a fatal intoxication case, and assessed various IS performance based on structural similarities and retention time.

Methods: A mid-teenager died from intoxication of personally imported dextromethorphan and Over-the-counter diphenhydramine. To quantify these drugs, we selected morphine-d₃, dihydrocodeine, diphenhydramine-d₃, mianserin-d₃, and diazepam-d₅ as alternative IS and evaluated. After selecting the most suitable IS, we quantified dextromethorphan and diphenhydramine concentrations in twelve plasma samples from the victim by liquid chromatography-tandem mass spectrometry.

Results: Recovery rates were 80.7-105.5%, except for morphine-d₃ (47.8%) and dihydrocodeine (64.8%). Matrix effects were 75.7-103.2%. The intra-day accuracies and precisions were 86.4-119.5% and 0.27-12.2%, respectively. The inter-day accuracies were 81.2-119.8%, and the precisions were 0.80-9.44%. The validation study showed that diphenhydramine-d₃ was the most suitable IS. Finally, plasma concentrations of dextromethorphan and diphenhydramine were 3.74-10.3 µg/mL and 15.6-52.9 µg/mL, respectively.

Conclusions: The concentrations of both drugs in plasma samples were estimated to cause death. When using an alternative IS, a validation study is needed to select the optimal IS.

Purpose: Comparison of the impurity removal efficiencies of the deproteinization and Quick, Easy, Cheap, Effective, Rugged, Safe (QuEChERS) methods, which are pretreatment methods for drug analysis adopted by many forensic autopsy institutions, was performed.

Method: Residual cardiac blood samples were pretreated using deproteinization and QuEChERS methods. The residual amounts of total proteins, total lipids, glucose, galactose, electrolytes, and inorganic elements were measured. We also compared the recovery rates and matrix factors when using liquid chromatography/tandem mass spectrometry (LC/MS/MS).

Results: The residual rates of total proteins, total lipids, glucose, galactose, and electrolytes using the deproteinization method were 16%, 75%, 75%, 90%, and 91%, respectively. In contrast, the QuEChERS method showed 1.1%, 11%, 7.6%, 9.4%, and 20%, respectively. The amounts of Mg and Mn in QuEChERS increased compared with those before treatment, but other inorganic elements remained at 9.6-89% during deproteinization and 0.30-17% in the QuEChERS. The recovery rate of metformin was low in QuEChERS; however, no differences were observed in the recovery rates or matrix factors of the other 16 drugs between deproteinization and QuEChERS.

Conclusions: This study quantitatively demonstrated that QuEChERS is extremely efficient at removing impurities from blood compared with deproteinization methods. QuEChERS has poor recovery rates for highly polar drugs but does not prevent their detection. The QuEChERS method is superior to the deproteinization method, considering the load of impurities on the analytical instruments.

Purpose: The analysis of drug residues on some currencies is well-established in the literature. However, there is no published study describing the presence of drug residues on Turkish paper currency.

Methods: This study focused on the analysis of 14 drug residues present on 600 Turkish banknotes collected from three different cities: Ankara, Adana, and Istanbul. The banknotes underwent preparation by a non-destructive and straightforward extraction method using methanol. To investigate the extent of contamination a method was subsequently developed and validated for liquid chromatography triple quadrupole mass spectrometry analysis to detect and quantify the target analytes. The investigated substances included benzoylecgonine, cocaine, heroin, codeine, morphine, 6-monoacetylmorphine (6-AM), amphetamine, methamphetamine, 3,4-methylenedioxy-N-methamphetamine (MDMA), methyl 3,3-dimethyl-2-(1-(pent-4-en-1-yl)-1H-indazole-3-carboxamido)butanoate (MDMB-4EN-PINACA), N-[1-(aminocarbonyl)-2,2-dimethylpropyl]-1-butyl-1H-indazole-3-carboxamide (ADB-BUTINACA), tetrahydrocannabinol (THC), pregabalin, ketamine, and tramadol.

Results: The calculated mean concentrations per note were 475.5 ng cocaine, 660.7 ng methamphetamine, 220.4 ng benzoylecgonine, 36.5 ng ketamine, 46.0 ng amphetamine, 120.6 ng 6-AM, 22.9 ng morphine, 6.3 ng codeine, 107.4 ng THC, 1.3 ng MDMB-4en-PINACA, 1.1 ng ADB-BUTINACA and 65.9 ng MDMA. Our findings indicate that banknotes commonly circulated in the three cities were primarily contaminated with methamphetamine and cocaine.

Conclusions: This study highlights the prevalence of drug residues on banknotes and raises concerns about their potential impact. The contamination of Turkish currency with drug residues is a strong indication of the widespread use of banknotes in drug trafficking.