Forensic Toxicology最新文献

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: 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.

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: 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 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: 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: 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: 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: 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: 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.