Forensic Toxicology最新文献

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: 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: 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.

Purpose: Existing on-site color tests for cannabis (e.g., rapid Duquénois-Levine reagent, 4-aminophenol reagent) have insufficient specificity, especially in the point of distinguishing between Δ⁹-tetrahydrocannabinol (Δ⁹-THC) and the other cannabinoids. We developed a novel Δ⁹-THC-specific color test reagent.

Methods: Cannabinoid standards were dissolved in 0.5 mL of a 2-hydroxybenzaldehyde solution in acetonitrile (2 g/100 mL) in a glass test tube (solution A). Herbal samples (approximately 10 mg) were extracted using 0.5 mL of the 2-hydroxybenzaldehyde solution in a plastic homogenization vessel. The extract was poured into a glass test tube through a filter cap (solution B). Solutions A and B were mixed with hydrochloric acid (0.5 mL) and allowed to react for 30 s, after which the mixture was extracted with chloroform (1 mL).

Results: When observing the lower layer after the chloroform extraction, Δ⁹-THC and cannabidiol (CBD) exhibited bluish and orangish colors, respectively. In contrast, cannabinol (CBN) did not show any color. The performance of the reagent was evaluated using dried herbal cannabis (n = 58) and other herbal materials (n = 13), including CBD- and CBN-supplemented herbal materials (each n = 2). Only cannabis samples with Δ⁹-THC levels ≥ 0.14% exhibited a bluish or greenish-blue color.

Conclusions: The newly developed reagent shows good specificity for Δ⁹-THC. The new reagent has the potential to replace existing on-site color tests.

Purpose: Cytisine is the active ingredient in preparations used for smoking cessation. Its popularity is attributed to its low cost, efficacy, and low incidence of adverse effects. Additionally, its easy over-the-counter availability is also significant. This accessibility makes it a potential substance for use in suicidal attempts. The aim of this study was to develop a method for the determination of cytisine in biological material for use in clinical and forensic toxicology, and to apply this method in authentic cases.

Methods: Biological samples were subjected to liquid-liquid extraction using cytisine-d₄ as an internal standard. Analyses were performed using a Hydrophilic Interaction Liquid Chromatography (HILIC) column with the technique of ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry.

Results: For both matrices (blood and urine), the linear concentration range was 5-1000 ng/mL. The method met all validation requirements. The concentration of cytisine in a man taking it for smoking cessation in post-mortem materials was 21.4 ng/mL in blood, 958.9 ng/mL in urine, ca. 30 ng/mL in vitreous humor, and ca. 40 ng/mL in bile. In contrast, for a man with cytisine intoxication, the concentration was 174.6 ng/mL in blood and > 10,000 ng/mL in urine. In both cases, no N-methylcytisine was detected.

Conclusions: The developed method can be used for the determination of cytisine in post-mortem biological matrices as well as for clinical purpose. We presented the concentrations of cytisine in the post-mortem biological samples of a man taking cytisine for smoking cessation and of a man with suicidal cytisine poisoning.

Purpose: Fluvoxamine (FLV) has been used widely as an antidepressant agent belonging to the group of second-generation selective serotonin reuptake inhibitors. However, only one work on the human metabolism of FLV was reported in 1983, examining a human urine specimen, and tentatively identified nine metabolites. Therefore, in the present work, the metabolites of FLV were examined in the liver, bile, and urine from a human cadaver, and the metabolites produced in the human liver microsomes (HLMs) in vitro were also examined.

Methods: Metabolites in each matrix were treated altogether in a tube where impurities had been precipitated using acetonitrile. The identification and tentative quantification of metabolites in human specimens and HLMs were performed using liquid chromatography (LC)-high resolution mass spectrometry (MS), LC-tandem mass spectrometry (MS/MS) and LC-QTRAP- MS/MS.

Results: Eleven new metabolites designated as M1 to M11 were detected from human cadaver specimens and HLMs. M1 was produced after acetylation at the terminal NH₂ of FLV and was the most abundant metabolite in the liver and bile, but was the third abundant one in urine. M4 was produced after demethylation at the methoxy moiety of FLV, and was the most abundant metabolite in HLMs.

Conclusions: To our knowledge, this is the first report on the existence of eleven new metabolites (M1-M11) of FLV in HLMs, human liver, bile and urine. The present eleven metabolites may be useful for the identification of FLV in human samples both antemortem and postmortem.