Forensic Toxicology最新文献

Purpose: The recreational use of synthetic cannabinoids (SCs) by adolescents and adults has markedly increased in recent years. Previous studies demonstrated that exposure to SCs is associated with multiple adverse health effects. Nevertheless, little is known about the effects of these substances on male fertility. The current study aimed to investigate the toxicological effects of subacute exposure to synthetic cannabinoid AB-FUBINACA on male reproductive system in mice.

Methods: Adult male Balb/c mice received daily intraperitoneal injections of various doses of AB-FUBINACA (0.75, 1.5, and 3 mg/kg for 3 weeks). Using biochemical and molecular methodologies, the impact of AB-FUBINACA on serum levels of reproductive hormones, sperm viability as well as various parameters in testicular tissue were evaluated.

Results: Our findings demonstrated that AB-FUBINACA induces dose-dependent reduction in testosterone levels in the serum, but not in follicle-stimulating hormone or luteinizing hormone. AB-FUBINACA treatment also causes a significant dose related decrease in sperm viability. These findings were associated with higher level of oxidative stress (GP91 expression and malondialdehyde level) and elevated expression of key regulators of apoptosis (Bax and caspase-3) as well as reduced expression of mitochondrial respiratory chain complexes SDHB (II), UQCRC2 (III), and ATP5a (V) in the testicular tissue.

Conclusion: From these findings, it can be concluded that exposure to AB-FUBINACA can interfere with the normal physiology and functioning of the male reproductive organs. Hence, gaining insight into the mechanisms by which SCs interfere with male fertility could guide future interventions and treatments.

Purpose: The abuse of 2-fluoro-2-oxo-phenylcyclohexylethylamine (2F-2-oxo-PCE), a dissociative anesthetic structurally related to phencyclidine (PCP) and ketamine, has recently increased in South Korea. This study presented the first forensic toxicological detection of 2F-2-oxo-PCE in autopsy cases and described a validated method for the simultaneous quantification of 2F-2-oxo-PCE, methylenedioxymethamphetamine (MDMA), methylenedioxyamphetamine (MDA), ketamine, and norketamine in postmortem blood.

Method: 2F-2-oxo-PCE and its metabolite, 2F-deschloronorketamine (2F-DCNK) were identified using gas chromatography-mass spectrometry (GC-MS). A liquid chromatography-tandem mass spectrometry (LC-MS/MS) with solid-phase extraction (SPE) was developed and validated for the simultaneous quantification of 2F-2-oxo-PCE, MDMA, MDA, ketamine and norketamine in blood.

Results: The validation parameters including linearity, accuracy, precision, matrix effect, and recovery were satisfactory. In postmortem blood samples, 2F-2-oxo-PCE concentrations ranged from 664 to 7911 ng/mL. Concurrent detection with MDMA and ketamine suggested possible polydrug use contributing to fatal outcomes.

Conclusions: The validated LC-MS/MS method is suitable for forensic applications and may enhance the toxicological profiling of emerging dissociative substances. The results can provide critical baseline data for interpreting 2F-2-oxo-PCE-related intoxications.

Purpose: Orexin receptor antagonists (ORAs) are a novel class of medications used in the treatment of insomnia. With increasing restrictions on benzodiazepine prescriptions, a rise in ORA overdose is expected. To enable the prediction of clinical severity and formulation of appropriate treatment strategies, we developed a rapid analytical method for detecting ORAs in plasma samples using a Monolithic solid-phase extraction (SPE) and gas chromatography-mass spectrometry (GC-MS).

Method: Extraction was performed using MonoTip C18. All steps from pretreatment to elution were conducted using centrifugation. Quantification was carried out using GC-MS with temperature-programmed analysis by positive ion electron ionization, using suvorexant-d₆ as the internal standard. The method was applied to plasma samples from actual ORA overdose cases to evaluate its practical applicability.

Result: The calibration curves demonstrated excellent linearity over the range of 10-2,000 ng/mL, with correlation coefficients of at least 0.9999. Reproducibility showed a coefficient of variation (CV) between 0.6% and 6.9%, and recovery rates were over 83%. ORA concentrations in overdose patient samples were successfully quantified using this method.

Conclusion: MonoTip C18 utilizes a reduced amount of solvent, thereby eliminating the need for evaporation-to-dryness steps. As a result, the entire procedure, encompassing SPE to GC-MS detection, can be completed within 40 min. This single protocol is applicable to all ORAs currently available in Japan and is suitable for both clinical and forensic toxicological applications.

Purpose: Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are direct metabolites of ethanol (EtOH) and sensitive biomarkers of alcohol consumption. However, despite extensive studies in Western populations, data on Japanese individuals are limited. This study characterized the pharmacokinetics of EtG and EtS in Japanese adults and evaluated the influence of dose, genetic polymorphisms, and habitual alcohol use.

Methods: Twenty-eight healthy Japanese adults received either 1.0 or 0.2 g/kg of pure EtOH (high or low dose). Whole blood and urine samples were collected for 24 h, and EtG and EtS were quantified using validated liquid chromatography-tandem mass spectrometry. Pharmacokinetic parameters were analyzed. The urinary excretion and recovery of EtG and EtS were estimated. Associations between genetic polymorphisms in alcohol-metabolizing and conjugation-related enzymes and Alcohol Use Disorders Identification Test (AUDIT) scores were also evaluated.

Results: EtG and EtS persisted longer than EtOH in both blood and urine. Both metabolites were excreted in the urine in a dose-dependent manner. Individuals carrying ALDH2*1/*2 showed a significantly higher urinary EtG formation rate than those carrying ALDH2*1/*1 (wild type). The AUDIT score showed a modest positive association with the urinary formation rate of EtS but not with EtG. The 24 h urine from high-dose participants exceeded international cutoffs, whereas that from low-dose participants was below the quantification limits.

Conclusions: EtG and EtS showed sustained detectability, and their urinary excretion was dose-dependent, indicating their utility as biomarkers of recent alcohol intake. These findings support their potential forensic applications of EtG and EtS in Japan.

Clinical trial registration: Japan Registry of Clinical Trials (jRCT), jRCT1070240083 (registered 2024-12-10).

Purpose: Some toxic plants have strong morphological similarities with edible wild plants. Therefore, poisoning often occurs due to accidental ingestion. It is important to identify poisonous plants in edible plant mixtures, even if the samples have been digested. In the present study, we developed a method for genus and species identification in mixed samples using massive parallel sequencing (MPS).

Methods: Veratrum oxysepalum and Colchicum autumnale are the most common causative plants of such accidents and their morphological features are similar to those of the edible Hosta sieboldiana. In this study, we used V. oxysepalum and C. autumnale as the target poisonous plant species. We developed and optimized an MPS analysis method for trnL and rbcL regions that are commonly used in plant species identification. Initially, DNA from poisonous plants (V. oxysepalum and C. autumnale) and edible plants (H. sieboldiana) were mixed in various ratios and analyzed using MPS. Next, we prepared cooked materials and simulated gastric contents from V. oxysepalum, C. autumnale, and H. sieboldiana and analyzed them using MPS.

Results: We detected both poisonous and edible plant DNA when mixed in equal amounts. Poisonous plants were also detected in cooked or simulated gastric acid content. These results suggest that our method can be used to identify the genera or species of plants present in cooked materials and simulated gastric contents.

Conclusions: These results indicate that MPS techniques are useful for the forensic analysis of plant materials.

Purpose: Forensics, along with the implementation of techniques from nanosciences, has brought about a change in the domain of forensic science and criminal investigation. It has brought about a positive change in the criminal investigation framework by making investigation more efficient and to-the-point.

Methods: This paper reviews the nanotechnology-based techniques that are introduced in forensic science to unravel the mysteries behind crimes as it highlights various kinds of nano-sized particles, nanodevices along with their applications that the forensic experts use to analyze the evidences collected from the crime scenes. Google scholar, PubMed, and Scopus search engines are used to select the related articles to write this review paper.

Results: DNA analysis, fingerprint detection, drug detection, explosive analysis, blood stain analysis, time since death estimation, and analysis of counterfeit documents are some of the sectors in which nanotechnology has made notable contributions.

Conclusion: As the paper unfolds, it makes sure that the readers get the taste of both the worlds, and that it helps them grasp the concept and idea behind the techniques used to make a change across the globe.

Purpose: Anabolic-androgenic steroids (AAS) enhance athletic performance, giving athletes an unfair advantage and disrupting fair competition. Banned in sports and listed by World Anti-Doping Agency, they require precise detection. This study aimed to develop a method using the transient matrix effect to improve AAS identification in biological samples.

Methods: Gas chromatography-tandem mass spectrometry (GC-MS/MS) method for determination of AAS samples was developed and validated. Biological samples were prepared using the QuEChERS technique.

Results: The optimised and validated method enhances AAS signals using high-boiling protectants. It ensures good linearity, low detection limits, and reliable precision. Optimal QuEChERS extraction and multiple reaction monitoring transitions in GC-MS/MS were evaluated, confirming applicability with blood plasma samples. The addition of a protectant to the analysed sample results in several notable effects. High-boiling protectants, such as polyethylene glycol (PEG-400), tetradecanoic acid (C14-COOH), n-tetradecylalcohol (C14-OH), and n-tetradecylamine (C14-NH₂), significantly enhance AAS's signal in blood plasma. This enhancement, however, is accompanied by a transient matrix effect induced by the protectants. PEG-400 produced the most substantial signal increase, with the response for nandrolone rising by as much as 912%.

Conclusions: The results demonstrate the potential offered by the utilisation of PEG-400 as a protectant to generate a transient matrix effect. The outcome of this process is an increased analytical signal from AAS in blood plasma, enabling their identification even at trace concentrations. The methodology developed and applied during the study can be used to reduce the detection limit of steroids and thus improve antidoping measures in sport.

Purpose: The aim of this work is to investigate whether precyclization of γ-hydroxybutyric acid (GABA) allows for increasing its gas chromatography (GC) signal, and if so, is it a more effective way to increase the signal of this compound than its silylation or methylation?

Methods: Gas chromatography-mass spectrometry (GC-MS) and GC with flame ionization detection (GC-FID) response to GHBA before and after silylation, methylation, and cyclization were compared. The impact of injector temperature on GHBA and γ-butyrolactone (GBL) signals was assessed. Fourier transformed infra-red spectroscopy was used to examine the formation of macromolecular derivatives in the injector.

Results: GHBA shows a lower GC signal than GBL due to partial polycondensation into a non-volatile polyester in the injector. Validation data were established for GHBA after each derivatization. Silylation and methylation reduced the limit of detection (LOD) by approximately 1.5- and 1.3-fold, respectively, whereas pre-cyclization led to at least a 4.6-fold decrease in LOD.

Conclusions: The present study elucidates the reasons behind the low GHBA signal observed in GC analysis and, consequently, supports the recommendation to perform pre-cyclization of this compound prior to analysis. Furthermore, the findings demonstrate that although signal enhancement of GHBA can be achieved through silylation or methylation, the most substantial increase is observed following its cyclization during sample preparation. The proposed in this paper cyclization procedure is both remarkably simple and highly effective, allowing for reliable quantification of this hydroxycarboxylic acid in a variety of matrices, including plasma, urine, wine, beer, and orange juice.

Purpose: A powder found at a fatality scene, labeled as the synthetic cathinone 3',4'-methylenedioxy-α-pyrrolidinohexiophenone (MDPHP) and most likely smoked using a crack pipe, was analyzed. The powder was identified as very potent synthetic cannabinoid ADB-BUTINACA/ADB-BINACA with a high purity (> 98%). This case highlights the risks associated with mislabeled novel psychoactive substances (NPS), particularly those purchased online.

Methods: The powder was analyzed using liquid chromatography-high resolution mass spectrometry (LC-HRMS), liquid chromatography-tandem mass spectrometry (LC-MS/MS), and nuclear magnetic resonance (NMR) spectroscopy. Comprehensive toxicological screening, including NPS, was performed in urine and blood. ADB-BUTINACA was quantified using a standard addition method on various post-mortem matrices: femoral and heart blood, urine, stomach content, bile fluid and liver tissue. Scalp hair was analyzed via external calibration to assess potential long-term exposure.

Results: ADB-BUTINACA concentrations were 34.5 ng/mL in femoral blood, 101 ng/mL in heart blood and 3.1 ng/mL in urine. Traces of MDMB-BUTINACA were found in the powder and hair, but not in other biological matrices. ADB-BUTINACA metabolites were detected in all biological matrices. MDPHP was found at low concentrations (< 2 ng/mL) in blood and urine but not in the powder, indicating prior cathinone use. A toxicological significance score (TSS) of 3 was assigned for this monointoxication with ADB-BUTINACA.

Conclusions: This case demonstrates fatal poisoning due to extremely high ADB-BUTINACA concentrations in post-mortem blood samples, emphasizing the severe risks associated with mislabeled substances. It underscores the importance of drug checking services to prevent poisonings and overdoses caused by highly potent NPS.