Quantitative analysis of volatile hydrocarbons (VHCs) in postmortem blood provides insight into the circumstances of fire-related deaths. Detection of specific VHCs can distinguish construction fires from those involving gasoline or kerosene. Although VHC concentrations have been examined in heart blood, their concentrations in other specimens remain unclear. We evaluated the correlations of carboxyhemoglobin and VHC concentrations among heart blood and femoral vein blood or various organ tissues to investigate the viability of alternative specimens for cases where heart blood cannot be collected. Heart blood, femoral vein blood, brain, liver, and muscle samples were collected from victims of fire-related death cases processed at our department. Seventeen VHCs were analyzed using headspace gas chromatography–mass spectrometry. In construction fire cases, carboxyhemoglobin concentrations in heart blood were positively correlated with those in femoral vein blood. Benzene, toluene, and styrene are commonly detected in fire-related deaths, and the concentrations of these compounds in heart blood were positively correlated with the corresponding concentrations in femoral vein blood. The concentrations in heart blood were significantly higher than those in femoral vein blood. Tissue concentrations were also positively correlated with those in heart blood. Among the samples, brain tissue had the highest concentrations. In gasoline- and kerosene-related fire cases, the VHC concentration patterns in the brain were similar to those observed in heart blood. These results suggest that brain tissue is an appropriate specimen for VHCs analysis and can be used when VHCs are undetectable in heart blood or heart blood is not available.
Hexahydrocannabinol (HHC) is an emerging semi-synthetic cannabinoid, but analytical data for all structurally confirmed stereoisomers of its hydroxylated metabolites remain limited. In this study, we synthesized all four stereoisomers of 8-hydroxyhexahydrocannabinol (8-OH-HHC) and comparatively characterized them, together with their trimethylsilyl (TMS) derivatives, by gas chromatography mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). A particular analytical advantage of the present work is that it includes LC-MS/MS data for (8S,9R)-8-OH-HHC, for which no standard-based analytical data was previously available. Four synthetic 8-OH-HHC stereoisomers were analyzed by GC-MS and LC-MS/MS. For GC-MS, both underivatized compounds and their TMS derivatives were examined. Retention behavior, electron ionization mass spectra, and LC-MS/MS transitions were compared among the stereoisomers. Under the tested GC conditions, the four stereoisomers and their TMS derivatives showed different retention times, although some differences were small and should be interpreted cautiously. Two pairs of stereoisomers exhibited similar electron ionization fragmentation patterns, whereas one pair showed differences in the relative abundances of selected ions. In addition to providing GC-MS data for all four underivatized stereoisomers, this study also provides GC retention and EI mass spectral data for all four TMS derivatives and a comparative evaluation of relative ion abundances among these closely related isomers. In LC-MS/MS, the stereoisomers showed partially overlapping transition patterns and closely eluting peaks under the tested conditions. These data provide a comparative analytical reference dataset for four structurally confirmed 8-OH-HHC stereoisomers and their TMS derivatives and may support future forensic studies of HHC-related compounds.
Xylazine, an α2-adrenergic agonist, is widely used in veterinary medicine for anesthesia, sedation, and analgesia. However, non-therapeutic use, overdose, and deliberate administration may pose serious health risks to animals, necessitating reliable detection methods for both clinical and forensic purposes. In this study, a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) method was developed and fully validated for the determination of xylazine and its metabolites in canine urine. The method exhibited excellent selectivity and linearity over the range of 0.05–100 ng/mL, with coefficients of determination (R²) ≥ 0.9989. Recovery values ranged from 70.3
Hair toxicological analysis enables the verification of long-term drug use monitoring and supports epidemiological studies. In recreational contexts, multi-analyte methods are crucial for accurately characterizing consumption patterns. The present work aimed to develop and validate an LC-MS/MS method for the analysis of 47 drugs of abuse and related compounds in hair samples. A 20 mg hair aliquot was incubated with methanol at 40 °C for 16 h. The supernatant was dried and injected into the LC-MS/MS system. The monitored substances were amphetamines, cocaine and related compounds, cannabinoids, opioids, hallucinogens, synthetic cathinones, synthetic phenethylamines, and synthetic cannabinoids. The method was validated in accordance with international recommendations and subsequently applied to 81 hair samples collected at Brazilian electronic dance music festivals. The method had limits of detection of 0.5-5 pg/mg and limits of quantification of 5–50 pg/mg. All compounds showed suitable linearity (R² > 0.9903). The precision assays showed CV
The reliable identification of amphetamine in degraded biological samples is often complicated by the presence of putrefactive amines, such as phenethylamine, which exhibit similar physicochemical properties and can interfere with chromatographic separation. This study aimed to systematically evaluate the conditions of gas chromatography–mass spectrometry (GC–MS), including derivatization methods and analytical column selection, to achieve effective separation and identification of amphetamine in the presence of phenethylamine. Various derivatization approaches, including acylation, alkoxycarbonylation, silylation, and pentafluorobenzylation, were examined in combination with non-polar, slightly polar, and semi-polar GC columns. The chromatographic behavior of amphetamine and phenethylamine, including the retention time, peak shape, and formation of mono- or di-substituted derivatives, was systematically assessed to optimize separation and mass spectral identification. Alkoxycarbonylation, silylation, and alkylation produced overlapping peaks, mixed derivatives, or unreacted compounds, thereby limiting their reliable identification. Acyl derivatization allowed for effective separation using non-polar and semi-polar columns, with the non-polar column minimizing peak tailing that is commonly caused by high phenethylamine concentrations. Differences in the elution order were observed between column types, and semi-polar columns provided satisfactory chromatographic resolution and high-quality mass spectra, consistent with spectral databases. Successful GC–MS identification of amphetamine in putrefied samples requires careful selection of the derivatization chemistry and GC column polarity. Acyl derivatization combined with an appropriate column choice enables reliable separation from phenethylamine, improves spectral quality, and enhances the evidential value of forensic analysis.
3-Chlorophenmetrazine (3-CPM) is a phenmetrazine analog that has been detected in new psychoactive substance (NPS) products sold on the market. The objective of this study was to perform an analytical characterization of 3-CPM identified in NPS products. In addition, 2-chlorophenmetrazine (2-CPM) and 4-chlorophenmetrazine (4-CPM), which are positional isomers of 3-CPM, were synthesized, and analytical methods for their differentiation were validated to prevent misidentification of the three CPM isomers. Samples were analyzed using liquid chromatography with photodiode array detection (LC/PDA), liquid chromatography-high resolution mass spectrometry (LC/HRMS), gas chromatography-mass spectrometry (GC/MS), gas chromatography-high resolution mass spectrometry (GC/HRMS), nuclear magnetic resonance (NMR) spectroscopy, and X-ray crystallography. Four NPS products sold between October 2022 and February 2023 were found to mainly contain trans-3-CPM, as determined by NMR analysis. Quantitative analysis showed that the powder and crystal products contained approximately 0.86–1.98 g of 3-CPM per package, whereas the tablet product contained approximately 0.10 g of 3-CPM per tablet. In chromatographic analyses, 2-CPM, 3-CPM, and 4-CPM were distinguishable based on retention times using octadecylsilyl silica gel (ODS) and phenyl columns for LC/PDA, and HP-5ms and CycloSil-B columns for GC/MS. Small differences were observed between UV spectra of 2-CPM/3-CPM and that of 4-CPM. Furthermore, 2-CPM produced a characteristic product ion in the LC/HRMS spectrum. The three positional isomers (2-, 3-, and 4-CPM) can be differentiated using LC and GC-based analytical methods. The analytical data presented in this study will facilitate the structural elucidation and accurate identification of novel phenmetrazine analogs in future investigations.
Misoprostol is a uterotonic agent widely used in obstetric and gynecological practice and is of forensic importance due to its involvement in pregnancy termination cases and related medico-legal evaluations. This study aimed to investigate the effects of misoprostol exposure on neural tube development during early embryogenesis using a chick embryo model. Early-stage chick embryos were exposed to different doses of misoprostol. Neural tube development was evaluated through morphometric, immunohistochemical, and molecular analyses. Morphological changes were assessed, while the expression of proliferating cell nuclear antigen (PCNA) and caspase-3 was examined immunohistochemically. In addition, the expression levels of selected neurodevelopment-related genes were analyzed. Morphometric assessments revealed neural tube closure defects and developmental delay in the misoprostol-exposed groups. Immunohistochemical analyses demonstrated decreased PCNA expression and increased caspase-3 expression compared with controls. Molecular analyses showed significant alterations in the expression of genes associated with neurodevelopment. Misoprostol exposure adversely affected neural tube development by disrupting cellular proliferation, apoptosis, and neurodevelopment-related gene expression during early embryogenesis. These findings suggest potential developmental risks associated with early embryonic exposure to misoprostol and may contribute to forensic and medico-legal evaluations of prenatal exposure cases.
PURPOSE:Flunitrazepam is a widely used hypnotic agent detected in many fatal intoxication and suicide cases. However, its concentration in blood during autopsy differs from that at the time of death; it is either undetected or detected at lower concentrations than expected in post-mortem blood. In this study, we evaluated interactions between flunitrazepam and human blood using liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry (LC-Q-Orbitrap-MS). METHODS:We measured flunitrazepam, 7-aminoflunitrazepam via reduction by aldehyde oxidase 1 (AOX1), and the degradation product of 7-aminoflunitrazepam via the Fenton reaction by hemoglobin (Hb) in human blood. In addition, we also detected flunitrazepam, 7-aminoflunitrazepam, and their degradation products in the blood of individuals who died from intentional ingestion of flunitrazepam. RESULTS:We found that flunitrazepam was reduced to 7-aminoflunitrazepam by AOX1 in blood, and 7-aminoflunitrazepam was oxidized by Hb via the Fenton reaction. The mass spectra obtained by LC-Q-Orbitrap-MS revealed the formation of 7-amino-5-(2-fluorocyclohexa-1,3-dien-1-yl)-3-hydroxy-1-methyl-1,3-dihydro-2H-benzo[e][1,4]diazepin-2-one (7-amino-3-hydroxyflunitrazepam) in the Hb/H2O2 reaction mixture containing 7-aminoflunitrazepam, as well as in the blood of some individuals who died from intentional ingestion of flunitrazepam. CONCLUSION:7-Amino-3-hydroxyflunitrazepam is the degradation product of flunitrazepam at various temperatures. The degradation is brought about by post-mortem changes via AOX1-mediated reduction and Hb-mediated Fenton reaction. Thus, 7-amino-3-hydroxyflunitrazepam may be a valuable biomarker for correcting blood flunitrazepam concentration at the time of death in clinical cases.
In response to the increasing abuse of etomidate-related substances in China, this study developed and validated an ultra-high performance liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of etomidate (ET), metomidate (MET), propoxate (PRO), isopropoxate (I-PRO), CF3-etomidate (CF3-ET), and their metabolite etomidate acid (ETA) in human nails. This method enabled the first investigation into their accumulation patterns in authentic samples, offering scientific support for forensic monitoring. A 20 mg portion of nail was cryogenically ground and extracted with methanol containing the internal standard. After vortex mixing, ultrasonication, and filtration, the extract was concentrated under nitrogen flow and analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry. The limits of quantification (LOQs) for the target substances in nail samples ranged from 1 to 10 pg/mg. The calibration curves exhibited good linearity over the concentration range from the LOQ to 1000 pg/mg. All other evaluated validation parameters proved satisfactory. The validated method was applied to authentic nail samples to investigate the accumulation profiles of ET and its analogs. Analysis of the samples revealed that 52 tested positive. These positive cases showed a distinct accumulation pattern dominated by the parent drugs, with ETA as the sole metabolite detected. This study developed and validated a reliable method for the simultaneous quantification of ET, its analogues, and its metabolite in human nails. Notably, it is the first to be applied to authentic samples to investigate their accumulation profiles, supporting the use of nail analysis in forensic toxicology and drug abuse monitoring.
In postmortem ethanol analysis in decomposed cases, simultaneously detected volatile organic compounds (VOCs) serve as indicators of postmortem ethanol production. Among these, 1‑propanol is the most widely used marker; however, it is sometimes absent even in markedly decomposed bodies, thus making interpretation difficult. This study aimed to comprehensively detect postmortem VOCs in femoral muscle samples collected at autopsy and to re-evaluate indicators of postmortem ethanol production. Among consecutive autopsy cases from a single institution, those involving antemortem intake of substances or pathological conditions that could increase VOCs were excluded. The cases were classified according to the degree of external decomposition. Femoral muscle samples were analyzed using headspace gas chromatography–mass spectrometry, and the detection profiles of VOCs and their relationship with ethanol were examined. Twenty-eight VOCs were identified in the femoral muscle, and both the detection frequency and number of detected compounds increased with the progression of decomposition. Among the major compounds detected, isovaleraldehyde, butyric acid, and 1-propanol showed weak correlations with ethanol. Ethanol was also detected in cases where these compounds were present. Contrastingly, all of these compounds were sometimes absent even in the markedly decomposed cases, indicating their limitations when used as single indicators. This study elucidated the characteristics of VOCs produced postmortem in the femoral muscle and demonstrated both the utility and limitations of indicators for evaluating postmortem ethanol production. These findings provide fundamental data that may improve the reliability of ethanol interpretation in autopsies.
In forensic cannabis investigations, it is important not only to prove that seized materials are cannabis or not but also to clarify the relationship between cannabis samples. This study aimed to establish a method for evaluating the similarity of the genetic background of cannabis samples using Inter-Simple Sequence Repeat (ISSR)-PCR analysis, a variant of DNA testing. Forty-eight cannabis samples from 24 strains were collected. ISSR-PCR was performed using 10 ISSR primers, and the resulting band patterns were used for grouping. After analyzing the characteristics of each primer using a binary classification method, two numerical indicators, the similarity score and total likelihood ratio, were calculated to comprehensively evaluate the similarity between samples and assess their discrimination performance. Cannabis samples were grouped into band patterns using ISSR-PCR, but no single primer classified all strains. The “similarity score” (0–10), based on primers showing the same band pattern across 10 ISSR-PCRs, averaged 1.69 between different strains, 4.34 between individuals of the identical strain, and 9.48 for the identical individual. The mean “total likelihood ratio” calculated from the positive and negative likelihood ratios for binary classification using 10 primers was 1.28 × 10⁰, 1.11 × 10⁵, and 9.79 × 10⁹, respectively. Each indicator demonstrated high discrimination performance in evaluating genetic similarity. This study established numerical criteria for estimating the genetic similarity between unknown cannabis samples based on ISSR-PCR results using 10 primers.
New LSD analogs are a newly emerging and basically under-researched group of new psychoactive substances (NPS). The emergence of a completely new group of NPS make it difficult to monitor intoxications with these compounds. Moreover, forensic toxicological evaluation often requires comprehensive information on changes in the concentration of NPS in biological specimens over time following their collection. In this study, 12 LSD analogs were analyzed to determine their stability in whole blood and dry blood spots (DBS). The samples were stored at room temperature (DBS), 4 °C (DBS and whole blood), and − 20 °C (whole blood). Analyses were performed using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry. The stability study lasted 365 days, monitoring both the degradation rate of analytes and formation of degradation products in vitro. Moreover, the effect of chemical structure on analyte stability was assessed. Results proved that the stability of analytes depended on both storage conditions and their chemical structure. Majority of compounds showed better long-term stability in DBS samples. Furthermore, N-deacylation of ALD-52, 1P-LSD, 1B-LSD, 1 V-LSD, 1cP-MiPLA, and 1P-MiPLA was observed, which was more pronounced for LSD derivatives than for MiPLA derivatives. Studies indicate a strong correlation between the stability of LSD analogs and both their chemical structure and storage conditions. DBS cards frequently provide equaled or enhanced stability compared to whole blood, which makes it advisable to collect biological material in both forms to help maximize analytical options in forensic and toxicological investigations.
The continuous emergence of NPS has posed a persistent challenge to forensic toxicology for more than a decade. Each year, previously unrecognized compounds appear on the illicit drug market, complicating analytical identification and increasing the risk of severe intoxications. The purpose of this study was to identify designer methaqualone analogs emerging in recent years and summarize available toxicological data in a comprehensive narrative mini review. Online databases including PubMed, Google Scholar and early warning systems were systematically searched for reports on newly emerging designer methaqualone analogs. In addition, online forums were reviewed to collect information from recreational drug-users on psychoactive effects, effective doses, and routes of administration. The review summarizes metabolism of designer analogs and all reported intoxication cases, including concentrations found. Analytical aspects relevant to forensic practice are highlighted i.a. mass spectrometric characteristics (GC-EI-MS and LC-MS/MS with QqQ and QTOF instruments), differentiation of isomers, and stability in biological fluids and DBS cards over 365 days. Five newly emerging compounds, mephenoqualone, PPQ, PPTQ, Cl-PPQ, and MCPPQ, were identified on the illicit market. These substances have 100-fold higher potency than methaqualone and were described on online forums in late 2025. Newly developed analogs are readily available online and represent serious and underrecognized threat due to their high potency and very low effective doses. Continued development of early warning systems and analytical methodologies is essential. Forensic toxicologists worldwide should be aware that these compounds may soon be encountered in seized materials and biological samples from intoxication cases.
The interest in therapeutic applications of tetrahydrocannabivarin (THCV) recently increased. For this reason, we validated an online extraction liquid chromatography- tandem mass spectrometry (LC-MS/MS) method to investigate the formation of urinary metabolites and understand potential cross-reactivity of THCV metabolites in cannabinoid immunoassays. Urine samples were obtained after oral administration of Δ8-THCV to healthy participants. The protocol was approved by the Advarra Institutional Review Board (Pro00059879; approved December 20, 2021) and the trial was registered on clinicaltrials.gov (NCT05210634). Urine samples were collected pre-dose and pooled 0-8 hours post-dose. Urine samples were extracted using a simple one-step protein precipitation procedure and the extracts analyzed using online trapping LC-MS/MS in positive multiple reaction monitoring mode. All compounds passed validation criteria in urine. In the clinical samples, 11-nor-9-carboxy-Δ8-THCV (Δ8-THCV-COOH) was the main metabolite detected before and after incubation with glucuronidases. Of the urine pooled 0-8 hours post-dose, 70 out of 80 were reported positive by a cannabinoid immunoassay targeting Δ9-THC-COOH, despite being negative for Δ9-THC-COOH and positive mainly for Δ8-THCV-COOH in the LC-MS/MS analysis. The major metabolites of Δ8-THCV in urine were Δ8-THCV-COOH, 11-hydroxy-Δ8-THCV and Δ9-THCV-COOH that were extensively glucuronidated and cross-react with immunoassay routinely used for toxicology testing resulting in false positive results for Δ9-THC exposure.
Diphenidine (DPD) is a dissociative novel psychoactive substance (NPS) structurally related to phencyclidine and ketamine. Although DPD is legally regulated in Japan and several other countries, analogues sharing the core scaffold are not comprehensively regulated. Therefore, it is possible that analogues with minor scaffold modifications may continue to emerge. This study examined the effects of methoxy or hydroxy substitution at the 4-position of DPD on its blood–brain barrier (BBB) penetration and dopamine release in the synaptic cleft. Using in vivo brain microdialysis in freely moving unanesthetized rats, DPD, 4-methoxydiphenidine (4MeO-DPD), and 4-hydroxydiphenidine (4OH-DPD) (20 mg/kg, i.p. each) were administered, and concentrations in the nucleus accumbens and plasma were quantified by liquid chromatography–mass spectrometry. Extracellular dopamine levels were determined by high-performance liquid chromatography with electrochemical detection. To investigate carrier-mediated BBB transport, verapamil (P-glycoprotein, P-gp, inhibitor) or diphenhydramine (organic cation transporter, OCT, inhibitor) was administered 1 h prior to each compound. DPD and its analogues showed distinct BBB penetration profiles, among which 4OH-DPD showed the highest brain concentrations and dopamine release. Verapamil but not diphenhydramine pretreatment significantly increased brain extracellular concentrations and prolonged elimination half-lives of all compounds, particularly 4MeO-DPD. P-gp inhibition elevated brain-to-plasma concentration ratios, indicating restricted BBB penetration by P-gp. The dopamine concentration profile was consistent with those observed for DPD and its analogues. This study demonstrates that 4MeO-DPD and 4OH-DPD strongly elicit dopamine release compared with DPD. These findings show that P-gp regulates BBB penetration, offering important insights for the toxicological risk assessment for newly emerging NPS.
Freshwater planarians provide a rapid and scalable biological model for detecting drug-induced neurobehavioral effects. This study evaluated whether automated behavioral profiling in Dugesia dorotocephala could distinguish phytocannabinoid and synthetic cannabinoid exposure based on organism-level motor responses. Planarians were acutely exposed to Δ⁹-tetrahydrocannabinol (Δ⁹-THC), AB-PINACA, MA-CHMINACA, A-796,260, or JWH-412 at concentrations of 5–60 µg/mL in artificial spring water containing PEG-400. Locomotion and posture were recorded for 5 min and analyzed using LabGym, a supervised deep-learning–based behavioral classification system that quantified gliding, headshake activity, and sustained C-shaped postures. Distinct compound-associated behavioral profiles were observed. Δ⁹-THC and JWH-412 produced marked suppression of total locomotion relative to pooled controls. AB-PINACA and MA-CHMINACA preserved overall movement volume but produced severe disruption of coordinated gliding accompanied by frequent abnormal postural states. A-796,260 produced comparatively mild effects on locomotor organization. These findings revealed separable behavioral patterns that were further resolved in a two-dimensional state space by PCA (91.4
Purpose Cocaine is one of the most widely abused drugs, and the analysis of seizures including drug profiling is a critical task for forensic practitioners. Street cocaine samples frequently contain additives, and they could hinder conventional drug analysis. In this study, a multivariate analysis method was made to rapidly estimate the composition and quantity of cocaine and its additives using Fourier transformation-infrared absorption (IR) spectroscopy and Raman spectroscopy. Methods IR and Raman spectra of standard cocaine samples and 21 additive samples were acquired. Training dataset was synthetically constructed by making linear combinations of the standard spectra with randomly generated weight and noises. Four different models of non-negative least square regression (NNLS), non-negative matrix factorization (NMF), non-negative elastic net (Enet) and multilayer perceptron (MLP), were constructed. The constructed models were applied to street samples whose components were previously characterized by conventional methods. Results Though all four methods gave good root-mean-square errors on the training dataset, application to street samples gave different results. NNLS provided the sparsest and the most appropriate results. MLP also provided similar level prediction capability. NMF could not retrieve the original standard spectra. Enet could not give sparse results and application to the street samples showed large fitting residuals. Based on these results, a decision procedure to predict the contents of cocaine seizure was constructed by the combination of NNLS and MLP. Conclusion Vibrational spectroscopy combined with multivariate analysis demonstrated significant potential for the rapid, semi-quantitative prediction of cocaine seizures with the information on diluents/adulterants.
Purpose Aripiprazole is an atypical antipsychotic agent with antidepressant properties, indicated for the treatment of schizo-phrenia, bipolar I disorder, and major depressive disorder, available only by prescription. The authors note that 3-trifluo-romethylphenylpiperazine (TFMPP) may be misattributed in aripiprazole-positive cases, due to its likely coelution with 1-(2,3-dichlorophenyl)piperazine (DCPP), one of aripiprazole's metabolites. Methods Biological matrices collected post-mortem (blood and vitreous humor) underwent liquid-liquid extraction with ethyl acetate in an alkaline medium (pH 9), followed by in-depth toxicological assessment via ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry. Results Comprehensive analysis using multiple reaction monitoring (MRM) and product ion scan modes confirmed the absence of TFMPP in biological samples positive for aripiprazole. Conclusions In toxicological analyses, particularly in forensic toxicology, utmost caution must be exercised when reporting and interpreting results, both in cases involving living subjects and post-mortem materials. It is recommended to monitor not only major metabolites but also minor ones. Analytical methods should be developed using certified reference standards, and in disputed cases, methodologies should be established that allow for verification of the contested results.
Purpose This study evaluated the applicability of segmental hair analysis using liquid chromatography-tandem mass spectrometry (LC/MS/MS) to determine the cause of death in highly decomposed corpses, where collection of urine or blood samples was impossible. Methods Multiple hair samples were collected from a highly decomposed female corpse and cut into 1-, 3-, and 10-mm segments. Qualitative LC/MS/MS analysis was performed on the hair root and shaft segments, followed by detailed quantitative analysis of the hair fragments. Results Qualitative analysis detected the prescribed drugs including zolpidem in all segments, indicating a distribution pattern of daily drug intake. Detailed segmental analysis revealed zolpidem concentrations ranging from 30 to 70 pg/mm in the root segment, and from 200 to 400 pg/mm in the other segments. These findings ruled out a large intake of zolpidem immediately prior to death. Similarly, no evidence of acute overdose with other prescribed drugs was identified. Conclusion This study demonstrated that even in decomposed corpses, where conventional biological samples are unavailable, high-sensitivity LC/MS/MS analysis of hair segments, including the root, enables accurate determination not only of the deceased's routine intake of sleep-inducing drugs but also of immediately pre-mortem drug intake.
To our knowledge, no previous reports have described the postmortem distribution of thiamethoxam in fatal cases. This study aimed to quantify thiamethoxam in biological fluids and solid tissues from a suicide involving ingestion of Actara® insecticide (10
