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).
{"title":"Forensic implications of ethyl glucuronide and ethyl sulfate pharmacokinetics in Japanese adults: the influence of dose, genetic polymorphisms, and habitual alcohol consumption.","authors":"Yuko Suefusa-Shimogori, Hirokazu Wakuda, Shinichi Nureki, Megumi Kai, Daisuke Sakamoto, Nao Mori, Tatsuji Fujisawa, Masaharu Narihara, Naoto Uemura","doi":"10.1007/s11419-025-00747-y","DOIUrl":"10.1007/s11419-025-00747-y","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p><p><strong>Clinical trial registration: </strong>Japan Registry of Clinical Trials (jRCT), jRCT1070240083 (registered 2024-12-10).</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"152-166"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145631390","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}
Pub Date : 2026-01-01Epub Date: 2025-12-08DOI: 10.1007/s11419-025-00748-x
Hitomi S Kikkawa, Kouichiro Tsuge
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.
{"title":"Identification of toxic plants from poisonous samples using massively parallel sequencing.","authors":"Hitomi S Kikkawa, Kouichiro Tsuge","doi":"10.1007/s11419-025-00748-x","DOIUrl":"10.1007/s11419-025-00748-x","url":null,"abstract":"<p><strong>Purpose: </strong>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).</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>These results indicate that MPS techniques are useful for the forensic analysis of plant materials.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"231-240"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145699845","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}
Pub Date : 2026-01-01Epub Date: 2025-06-26DOI: 10.1007/s11419-025-00731-6
Michal P Dybowski, Krystian Siwek
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.
{"title":"Application of the transient matrix effect for determination of anabolic-androgenic steroids in biological samples by GC-MS/MS.","authors":"Michal P Dybowski, Krystian Siwek","doi":"10.1007/s11419-025-00731-6","DOIUrl":"10.1007/s11419-025-00731-6","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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%.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"204-216"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12858534/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144495450","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}
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.
{"title":"Innovative applications of nanotechnology in enhancing forensic science investigations.","authors":"Asmita Podder, Agnishwar Girigoswami, Koyeli Girigoswami","doi":"10.1007/s11419-025-00734-3","DOIUrl":"10.1007/s11419-025-00734-3","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusion: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"19-36"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783906","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}
Pub Date : 2026-01-01Epub Date: 2025-09-13DOI: 10.1007/s11419-025-00738-z
Rafal Typek, Michal P Dybowski, Andrzej L Dawidowicz
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.
{"title":"Cyclization of γ-hydroxybutyric acid (GHBA) as a strategy to enhance its signal in gas chromatography analysis.","authors":"Rafal Typek, Michal P Dybowski, Andrzej L Dawidowicz","doi":"10.1007/s11419-025-00738-z","DOIUrl":"10.1007/s11419-025-00738-z","url":null,"abstract":"<p><strong>Purpose: </strong>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?</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"107-119"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12858631/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145052625","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}
Pub Date : 2026-01-01Epub Date: 2025-11-27DOI: 10.1007/s11419-025-00746-z
Annette Zschiesche, Nadine Theofel, Stefan Braukmüller, Edwin Ehrlich, Martin Jasyk, Maximilian Methling, Michael Tsokos, Stefan Scholtis, Laura M Huppertz, Volker Auwärter
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.
{"title":"Deadly confusion of novel psychoactive substances: fatal outcome of ADB-BUTINACA mislabeled as 3',4'-methylenedioxy-α-pyrrolidinohexiophenone.","authors":"Annette Zschiesche, Nadine Theofel, Stefan Braukmüller, Edwin Ehrlich, Martin Jasyk, Maximilian Methling, Michael Tsokos, Stefan Scholtis, Laura M Huppertz, Volker Auwärter","doi":"10.1007/s11419-025-00746-z","DOIUrl":"10.1007/s11419-025-00746-z","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"257-270"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12858572/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145631488","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}
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.
{"title":"Development of a rapid targeted and non-targeted analysis method for etomidate and its structural analogues by ambient flame ionization mass spectrometry.","authors":"Meiting Lin, Miao Zhang, Zhen Zhang, Ping Xiang, Yunli Zhao, Junbo Zhao","doi":"10.1007/s11419-025-00737-0","DOIUrl":"10.1007/s11419-025-00737-0","url":null,"abstract":"<p><strong>Purpose: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"96-106"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144948462","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}
Purpose: Recently, a number of abuse and intoxication cases involving imidazole-derived GABA agonists etomidate and its analogs have been reported in China. During our recent screening of e-cigarette liquids, we encountered two novel etomidate analogs that have not previously been reported. This study aims to present the analytical procedures used to identify these compounds, along with detailed data obtained under various instrumental conditions.
Methods: Identification of the substances of concern was carried out using various instruments, including gas chromatography-mass spectrometry (GC-MS), headspace gas chromatography (HS-GC), liquid chromatography-high-resolution mass spectrometry (LC-HRMS), and liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Results: Analysis of the e-cigarette samples of concern using GC-MS, LC-HRMS, and LC-MS/MS yielded mass spectra, product ion mass spectra, and high-resolution mass spectra, which allowed structural elucidation. HS-GC analysis of hydrolyzed alcohols provided further insights into the substituent groups linked to imidazole core. Finally, the compounds were identified as butomidate and trifluoro-etomidate (tf-etomidate) through comparison with reference standards. Product ion mass spectra of the reference standards at various collision energies were obtained by LC-MS/MS, which also matched those of the target compounds.
Conclusion: This study demonstrated the ion spectra of butomidate, sec-butomidate and tf-etomidate as obtained by GC-MS, and their product ion spectra under varying collision energies by LC-MS/MS. These findings enabled the identification of the compounds in the dubious e-cigarette liquids. To our knowledge, this is the first report detailing qualitative procedures and associated instrumental data for the identification of butomidate and tf-etomidate.
{"title":"Identification of two novel imidazole-derived GABA agonists butomidate and tf-etomidate in e-cigarette liquids.","authors":"Xiaolong Zhang, Yaqing Li, Jinlei Liu, Ziyi Pan, Yuxuan Chen, Mengchao Wang, Yinyin Dai, Kundi Zhao, Jie Gu, Huimin Zhang, Shengnan Zhang, Amin Wurita, Koutaro Hasegawa","doi":"10.1007/s11419-025-00732-5","DOIUrl":"10.1007/s11419-025-00732-5","url":null,"abstract":"<p><strong>Purpose: </strong>Recently, a number of abuse and intoxication cases involving imidazole-derived GABA agonists etomidate and its analogs have been reported in China. During our recent screening of e-cigarette liquids, we encountered two novel etomidate analogs that have not previously been reported. This study aims to present the analytical procedures used to identify these compounds, along with detailed data obtained under various instrumental conditions.</p><p><strong>Methods: </strong>Identification of the substances of concern was carried out using various instruments, including gas chromatography-mass spectrometry (GC-MS), headspace gas chromatography (HS-GC), liquid chromatography-high-resolution mass spectrometry (LC-HRMS), and liquid chromatography-tandem mass spectrometry (LC-MS/MS).</p><p><strong>Results: </strong>Analysis of the e-cigarette samples of concern using GC-MS, LC-HRMS, and LC-MS/MS yielded mass spectra, product ion mass spectra, and high-resolution mass spectra, which allowed structural elucidation. HS-GC analysis of hydrolyzed alcohols provided further insights into the substituent groups linked to imidazole core. Finally, the compounds were identified as butomidate and trifluoro-etomidate (tf-etomidate) through comparison with reference standards. Product ion mass spectra of the reference standards at various collision energies were obtained by LC-MS/MS, which also matched those of the target compounds.</p><p><strong>Conclusion: </strong>This study demonstrated the ion spectra of butomidate, sec-butomidate and tf-etomidate as obtained by GC-MS, and their product ion spectra under varying collision energies by LC-MS/MS. These findings enabled the identification of the compounds in the dubious e-cigarette liquids. To our knowledge, this is the first report detailing qualitative procedures and associated instrumental data for the identification of butomidate and tf-etomidate.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"47-60"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144607898","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}
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.
用途:Δ9-Tetrahydrocannabinolic酸- a (Δ9-THCA-A)是大麻中Δ9-tetrahydrocannabinol的前体。在这里,考虑到适用于普通法医实验室,我们开发了一种新的隔离方法Δ9-THCA-A不需要特殊设备。方法:用乙腈提取干大麻花粉(2g)。提取液经石墨碳粉处理去除叶绿素后,用甲醇代替溶剂。甲醇溶液用氢氧化钠水溶液稀释,然后用正己烷/乙酸乙酯(7:1,v/v)的混合物洗涤。剩余水层用乙酸酸化,用正己烷/乙酸乙酯混合物萃取。提取液采用硝酸银-硅胶柱层析纯化。结果:分离得到的最终产物为淡米色固体。将三甲基硅基化后的气相色谱/质谱(GC/MS)分析结果与具有正品标准的高效液相色谱-紫外检测(HPLC-UV)分析结果进行比较,确定最终产品为Δ9-THCA-A。经1h -核磁共振波谱、GC/MS、HPLC-UV等方法验证,最终产物纯度高。结论:该分离方法不需要特殊的纯化设备,如闪蒸层析系统或带组分收集器的高效液相色谱系统,可获得高纯度的Δ9-THCA-A,这在法医实验室中并不常见。这种方法将对许多难以获得商业Δ9-THCA-A作为标准的法医实验室有用。
{"title":"Development of a novel isolation method for Δ<sup>9</sup>-tetrahydrocannabinolic acid-A from cannabis suitable for forensic laboratories.","authors":"Kenji Tsujikawa, Yuki Okada, Tadashi Yamamuro, Kenji Kuwayama, Tatsuyuki Kanamori, Yuko T Iwata","doi":"10.1007/s11419-025-00742-3","DOIUrl":"10.1007/s11419-025-00742-3","url":null,"abstract":"<p><strong>Purpose: </strong>Δ<sup>9</sup>-Tetrahydrocannabinolic acid-A (Δ<sup>9</sup>-THCA-A) is a precursor of Δ<sup>9</sup>-tetrahydrocannabinol in cannabis. Here, considering applicability to ordinary forensic laboratories, we developed a novel isolation method for Δ<sup>9</sup>-THCA-A without the need for special equipment.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>The isolation procedure gave a pale beige solid as the final product. The final product was identified as Δ<sup>9</sup>-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 <sup>1</sup>H-nuclear magnetic resonance spectroscopy in addition to GC/MS and HPLC-UV.</p><p><strong>Conclusions: </strong>This novel isolation method gave highly pure Δ<sup>9</sup>-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 Δ<sup>9</sup>-THCA-A as a standard.</p>","PeriodicalId":12329,"journal":{"name":"Forensic Toxicology","volume":" ","pages":"224-230"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145205975","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}