Forensic Toxicology最新文献

{"title":"An Ultra-High performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of etomidate, analogues, and metabolites in human nail: Unveiling a unique accumulation profile.","authors":"Pinting Zhang, Kaiyu Wang, Xinyi Yan, Dan Zhang, Jiale Bai, Zixia Shao, Yihan Liu, Zhenjiang Liao, Yunde Tang, Hao Zeng, Na Jing, Qiuping Huang, Hongxian Shen, Bin Di, Fang Yan, Tao Lu","doi":"10.1007/s11419-026-00776-1","DOIUrl":"https://doi.org/10.1007/s11419-026-00776-1","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusion: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2026-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148276320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a similarity discrimination method for cannabis samples using Inter-Simple Sequence Repeat-PCR.","authors":"Tadashi Yamamuro, Yuki Okada, Hiroki Segawa, Kenji Kuwayama, Kenji Tsujikawa, Tatsuyuki Kanamori, Yuko T Iwata","doi":"10.1007/s11419-026-00775-2","DOIUrl":"https://doi.org/10.1007/s11419-026-00775-2","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>This study established numerical criteria for estimating the genetic similarity between unknown cannabis samples based on ISSR-PCR results using 10 primers.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148257952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Volatile organic compounds (VOCs) in femoral muscle from forensic autopsy cases: exploring potential indicators of postmortem ethanol production: Part I - comprehensive profiling of postmortem generated VOCs.","authors":"Natsuki Ikematsu, Koji Yonemoto, Kenji Ninomiya, Kazumichi Kakazu, Maki Fukasawa, Mio Takayama, Aya Matsusue","doi":"10.1007/s11419-026-00774-3","DOIUrl":"https://doi.org/10.1007/s11419-026-00774-3","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148257880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reinventing quaaludes: a comprehensive review of designer methaqualone analogs.","authors":"Olga Wachełko, Kaja Tusiewicz, Marcin Zawadzki, Paweł Szpot","doi":"10.1007/s11419-026-00773-4","DOIUrl":"https://doi.org/10.1007/s11419-026-00773-4","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148249886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative stability assessment of novel LSD analogs in dry blood spots and blood using UHPLC-MS/MS: implications for forensic applications.","authors":"Kaja Tusiewicz, Paweł Szpot, Marcin Zawadzki","doi":"10.1007/s11419-026-00770-7","DOIUrl":"https://doi.org/10.1007/s11419-026-00770-7","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2026-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148249880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Treatment with rapamycin prevents induction and expression of locomotor sensitization to synthetic cathinone 3,4-methylenedioxypyrovalerone (MDPV) in mice.","authors":"Jakub Wojcieszak, Katarzyna Kuczyńska, Jolanta B Zawilska","doi":"10.1007/s11419-025-00749-w","DOIUrl":"10.1007/s11419-025-00749-w","url":null,"abstract":"<p><strong>Purpose: </strong>3,4-Methylenedioxypyrovalerone (MDPV) is a potent psychostimulant substance endowed with addictive properties. As mammalian target of rapamycin (mTOR) mediates neuroadaptive changes responsible for development of addiction, the current study evaluated whether rapamycin, a potent and selective inhibitor of mTOR, prevents induction and expression of behavioral sensitization in mice treated with MDPV.</p><p><strong>Methods: </strong>Locomotor sensitization was used as an animal model of early phase of addiction. C57BL/6JRj mice were treated with rapamycin before administration of MDPV during the induction phase of sensitization, or during the final 5 days of the withdrawal. Sensitization was assessed based on the measurement of locomotor activity after treatment with MDPV.</p><p><strong>Results: </strong>Rapamycin administered on days 1-7 inhibited induction of sensitization characterized by increased horizontal and vertical locomotor activity on day 7 compared to day 1. Additionally, when given during the withdrawal from MDPV, rapamycin blocked expression of sensitization, defined as augmented response to MDPV on day 21 compared to day 1.</p><p><strong>Conclusions: </strong>Abolishment of locomotor sensitization to MDPV by rapamycin suggests that neuroadaptive changes underlying this phenomenon are dependent on the mTOR signaling and warrants further research on possible application of mTOR inhibitors in treatment of addiction.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"141-151"},"PeriodicalIF":2.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12858557/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145631481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of toxicokinetic profiles of methamphetamine and its metabolites at toxic and therapeutic doses in mice.","authors":"Misato Ishida, Asuka Kaizaki-Mitsumoto, Satoshi Numazawa","doi":"10.1007/s11419-025-00753-0","DOIUrl":"10.1007/s11419-025-00753-0","url":null,"abstract":"<p><strong>Purpose: </strong>Methamphetamine (MA) is widely abused worldwide and has long been a major social concern. To provide basic information for comparing MA toxicokinetics in humans, we analyzed changes in the serum and urinary concentrations of MA and its metabolites in mice and compared the toxicokinetic profiles at the toxic dose with those at the therapeutic dose.</p><p><strong>Methods: </strong>Mice were administered therapeutic (1.5 mg/kg) or toxic (15 mg/kg) doses and blood and urine samples were collected. The serum concentrations of MA and its metabolite amphetamine (AMP), as well as the urinary concentrations of MA and its metabolites-AMP, p-hydroxymethamphetamine (OHMA), p-hydroxyamphetamine (OHAMP), and norephedrine (NEP) -were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS).</p><p><strong>Results: </strong>The serum AUC_0-24 values for MA and AMP were approximately 16 and 41 times higher, respectively, in the toxic dose group than those in the therapeutic dose group. The urinary AMP and OHAMP excretion levels were approximately 3.6 and 2.2 times higher, respectively, in the toxic dose group. The urinary [AMP] / [MA] ratio at all collected points and [AMP] / [OHMA] ratio in the 0-24 h sample were significantly higher in the toxic than in the therapeutic dose group.</p><p><strong>Conclusions: </strong>The results obtained from this study suggest that the metabolism of AMP to OHAMP and NEP was saturated during intoxication. Furthermore, the determination of whether a toxic dose had been administered within 24 h or after 24 h would be possible through a joint evaluation of the urinary [AMP] / [MA] and [AMP] / [OHMA] ratios.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"192-203"},"PeriodicalIF":2.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145755543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The determination of the novel insomnia medication lemborexant using a validated liquid chromatography-tandem mass spectrometry method, and its quantitation in clinical and forensic samples.","authors":"Brian Waters, Masato Masuda, Aya Matsusue, Masayuki Kashiwagi, Hikaru Hori, Kazuya Ikematsu, Shin-Ichi Kubo","doi":"10.1007/s11419-025-00733-4","DOIUrl":"10.1007/s11419-025-00733-4","url":null,"abstract":"<p><strong>Purpose: </strong>This study aims to detail the identification, confirmation, and quantitation of lemborexant from clinical and postmortem specimens using a validated LC-MS/MS method. Additionally, it investigates the tissue distribution of lemborexant in several postmortem cases.</p><p><strong>Methods: </strong>Lemborexant was isolated from the plasma of hospital patients or the postmortem specimens of forensic autopsy cases. Extraction from 0.1 mL or 0.1 g of sample was achieved by a modified QuEChERS protocol. The analysis was performed by liquid chromatography-tandem mass spectroscopy (LC-MS/MS). The quantitation method was validated in whole blood using internationally accepted parameters based on ANSI/ASB Standard 036. A total of 8 clinical samples and 13 forensic autopsy cases were analyzed by the validated method.</p><p><strong>Results: </strong>In clinical cases, lemborexant concentrations in plasma ranged from 2.7 to 225 ng/mL. Lemborexant concentrations in the blood of forensic autopsy cases ranged from below the lower limit of quantitation (2 ng/mL) to 276 ng/mL. The highest postmortem concentrations were found in liver, adipose tissue, pancreas, and kidney. The method demonstrated high recovery rates and precision, with no significant matrix effects or interferences from other drugs.</p><p><strong>Conclusions: </strong>The validated LC-MS/MS method proved effective for detecting and quantifying lemborexant in both clinical and forensic autopsy samples. The study highlights the importance of monitoring lemborexant and other dual orexin receptor antagonists (DORAs) in forensic investigations to understand their pharmacokinetics and potential toxicological effects.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"61-71"},"PeriodicalIF":2.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144648972","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Toxicological findings in suspected drugging incidents in Denmark from 2022 - 2024.","authors":"Charlotte Uggerhøj Andersen, Sys Stybe Johansen, Karen Rygaard, Simon Kjær Hermansen, Brian Schou Rasmussen, Jørgen Bo Hasselstrøm","doi":"10.1007/s11419-025-00752-1","DOIUrl":"10.1007/s11419-025-00752-1","url":null,"abstract":"<p><p>PURPOSE: Suspicions of drugging – the covert administration of psychoactive substances – have been increasing in Denmark. Our aim was to identify risk factors and characterize the substances detected in suspected drugging incidents. METHODS: We reviewed toxicological results and police-provided information for all subjects referred for forensic toxicological examination in Denmark between April 2022 and June 2024 due to suspected drugging. RESULTS: Three-hundred-and-seventy-three subjects were examined, of which 280 (75%) were female. Median [interquartile range] age was 23 [19–29] years. Median time from suspected incident to blood sampling was reported in 223 subjects, and was 18 [11–29] hours. We detected psychoactive drugs in 244 subjects (65%); most frequently ethanol (n = 179, 48%). In samples taken within 12 h, the median blood and urine alcohol concentration was 1.0 [0.5–1.5] ‰ (n = 58) and 1.7 [1.0-2.2] ‰ (n = 50), respectively. Central stimulants, antidepressants, and cannabis were present in 43 (11%), 34 (9%), and 25 (7%) subjects, respectively. Self-intake was reported for 82 subjects (22%). Among these, psychoactive drugs other than ethanol, which were not reported as self-intake, were detected in 20 (24%), most frequently central stimulants, antidepressants, and cannabis. However, in eight (10%), benzodiazepines, opioids, or sedating antihistamines not explained by self-intake were detected. Blackout was the most frequently reported symptom, noted by 45 (61%) out of 74 subjects, for whom symptom data were available. CONCLUSIONS: Risk factors for experiencing suspected drugging included young age, female sex, and exposure to alcohol, antidepressants and/or central stimulants. The detected drugs mostly reflected frequently used drugs in society.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"167-176"},"PeriodicalIF":2.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145755651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inactivation of calcium Ion signaling in neuronal SK-N-SH cells with development of resistance to a designer drug α-pyrrolidinooctanophenone.","authors":"Yuji Sakai, Yoshifumi Morikawa, Toshihiro Matsumura, Shunsuke Jimbo, Koichi Suenami, Gento Yamashita, Atsushi Nagai, Tomomi Michiue, Akira Ikari, Toshiyuki Matsunaga","doi":"10.1007/s11419-025-00751-2","DOIUrl":"10.1007/s11419-025-00751-2","url":null,"abstract":"<p><strong>Purpose: </strong>The cellular mechanisms underlying tolerance development to psychostimulant-induced neurotoxicity remain poorly understood. This study investigated these mechanisms using pyrrolidinophenone derivatives (PPs), potent amphetamine-type stimulants with strong dopaminergic activity and high cytotoxicity, aiming to establish a neuronal tolerance model and to explore adaptive processes relevant to substance use disorder.</p><p><strong>Methods: </strong>Human SK-N-SH neuronal cells were chronically exposed to α-pyrrolidinooctanophenone (α-POP) to prepare the drug-resistant cell line, SH/POP. Transcriptomic profiling was performed to identify gene expression alterations associated with tolerance development.</p><p><strong>Results: </strong>Cell sensitivity assay showed that SH/POP cells can survive at lethal concentrations (> 40 μM) of α-POP. RNA sequence analysis of the resistant cells identified alterations in 1,298 differentially expressed genes and the gene ontology analysis surmised an upregulation of calcium-binding-related genes. The development of α-POP resistance reduced the basal Ca²⁺ concentration and suppressed the caspase-3 activation elicited by the drug. Additionally, the development down-regulated the phosphorylation of a transcription factor cAMP response element-binding protein (CREB) and expressions of CREB-target genes, Fos proto-oncogene AP-1 transcription factor subunit and neurotensin. Furthermore, constitutive activation of endoplasmic reticulum stress responses and selective enhancement of trypsin-like proteasome activity in SH/POP cells were detected.</p><p><strong>Conclusions: </strong>Resistance development of neuronal cells to PPs is ascribable to suppression of calcium-dependent apoptosis and CREB signaling, and constitutive activation of endoplasmic reticulum stress responses. The SH/POP cell line represents a novel in vitro model to study molecular adaptations to psychostimulant toxicity and provides insights into neuroadaptive mechanisms underlying substance use disorder.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"177-191"},"PeriodicalIF":2.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145755526","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}