Journal of analytical toxicology最新文献

{"title":"Comparison of a Comprehensive Liquid Chromatography-Quadrupole Time-of-Flight-Mass Spectrometry (LC-QTOF-MS) Screen in Whole Blood with Conventional Immunoassay-Based Techniques.","authors":"Jessica Ayala, Sarah Kerrigan","doi":"10.1093/jat/bkaf054","DOIUrl":"https://doi.org/10.1093/jat/bkaf054","url":null,"abstract":"<p><p>Traditional immunoassay (IA)-based drug screens are limited in their scope of analysis and specificity. Their reliance on the cross-reactivity of antibody reagents is a limiting factor, particularly in light of the emergence of new therapeutics, emerging drugs, and new psychoactive substances (NPS). High resolution mass spectrometry (HRMS)-based techniques can offer improved versatility, specificity, and increase the scope of analytical testing. In this study, a validated HRMS screening procedure was used to reanalyze adjudicated blood samples previously tested by immunoassay. IA methods employed enzyme-linked immunosorbent assay (ELISA) and enzyme multiplied immunotechnique (EMIT®). The comprehensive HRMS screen utilized supported liquid extraction (SLE) and liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). Specimens previously tested by IA were reanalyzed using the HRMS screen following long term storage. The LC-QTOF-MS toxicology screen produced an additional 709 positive drug findings (67 compounds) among a population of 919 previously analyzed blood specimens. This study highlights the analytical benefits of MS-based toxicological screening and the advantages of data acquisition modalities that permit retrospective data interrogation.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144274882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Incorporation of an Environmentally Friendly Method for the Detection of Fentanyl and its Analogs in Oral Fluid.","authors":"Cynthia Coulter, Campbell Coulter, Jonah Gonzales, Christine Moore","doi":"10.1093/jat/bkaf053","DOIUrl":"https://doi.org/10.1093/jat/bkaf053","url":null,"abstract":"<p><p>Oral fluid is considered a favorable matrix for the identification of drug intake mainly because of its simple, observed, non-invasive collection. Fentanyl and fentanyl analog use, misuse, overdose, and deaths are currently occurring at an alarming rate in the USA. The law enforcement community, the Food and Drug Administration (FDA) and the Centers for Disease Control (CDC) are all keenly aware of the urgency in addressing an unmet public health need to identify opioid overdose in individuals as rapidly as possible. As part of a National Institute of Justice grant, the present study was intended to develop and validate an environmentally friendly, rapid, sensitive quantitative method using liquid chromatography coupled to tandem mass spectral detection (LC-MS/MS) for fentanyl and fentanyl analogs in oral fluid collected using the nform rapid test device. Oral fluid samples were subjected to liquid-liquid extraction incorporating bio-renewable solvents where possible, reducing the environmental footprint of the assay. A buffer/salt free mobile phase was employed consisting of 0.1% formic acid in water (95%): 0.1% formic acid in methanol (5%) at a flow rate of 0.8 mL/min; the run time was 4.5 minutes, again reducing environmental impact in terms of salt and solvent usage. The method included fentanyl, 4-anilino-N-phenethylpiperidine; (4-ANPP; desproprionyl fentanyl), acetyl fentanyl, carfentanil, p-fluorofentanyl, valeryl fentanyl, p-fluorobutyrylfentanyl, furanyl fentanyl and benzoyl fentanyl as well as xylazine, which is often detected with fentanyl. The method was validated according to ANSI/ASB 036 (2019) Standard Practices for Method Validation in Forensic Toxicology.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144274883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Changes in blood cannabinoid concentrations over multiple collection times in driving under the influence of drugs casework.","authors":"Brianna L Peterson, Meaghan R Hessler","doi":"10.1093/jat/bkaf052","DOIUrl":"https://doi.org/10.1093/jat/bkaf052","url":null,"abstract":"<p><p>Δ9-Tetrahydrocannabinol (THC) is the most frequently used illicit drug in the world, yet interpretation of THC concentrations in driving under the influence of drug (DUID) cases is difficult due to possible residual THC concentrations. This study determined the concentrations of cannabinoids in blood collected across multiple time points from drivers in suspected impaired driving cases to evaluate if changes in concentrations over time can provide clarification on time of cannabis use. This study examined cannabinoid-positive DUID cases reported from January 2019 to December 2023 to identify those that tested multiple blood draws. Thirty-five cases were identified that had multiple blood draws for a total of 81 different samples with collection times ranging from 00:32 hours to 12:42 hours between incident and blood draw. Cannabinoid testing was performed using a liquid chromatography-tandem mass spectrometry analysis with reporting limits of 1.0, 5.0, and 0.5 ng/mL for 11-hydroxy-THC (11-OH-THC), 11-nor-9-carboxy-THC (THC-COOH), and THC, respectively. THC concentrations (n = 81) ranged from 0.74-40 ng/mL. Eleven samples had an increase in THC concentration at a later collection time point. 11-OH-THC concentrations (n = 60) ranged from 1.0-16 ng/mL. THC-COOH concentrations (n = 81) ranged from 7.1-470 ng/mL. The results of this study underscore the difficulty in interpretation and drawing conclusions regarding time of cannabis use, even when multiple samples are obtained from the same subject over time from a single incident.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144247990","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LC-MS/MS determination of the novel fentanyl analog, ortho-methylfentanyl, in drug-related toxicity casework. Concentrations in ligated femoral blood.","authors":"C N Chatterton, R P Handy, G K Shoemaker","doi":"10.1093/jat/bkaf050","DOIUrl":"https://doi.org/10.1093/jat/bkaf050","url":null,"abstract":"<p><p>The purpose of this study was to develop and validate an analytical method to chromatographically separate, identify and quantify ortho-methylfentanyl (o-methylfentanyl) in postmortem blood. A combination of simple protein precipitation with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was utilized to facilitate chromatographic separation of similar fentanyl analogs, including both meta (m-) and para (p-) methylfentanyl. The analytical range was 1 to 200 ng/mL; the method was validated in accordance with ANSI/ASB Standard 036. In addition to providing details of the validated analytical method, this study details the results of the analysis of one hundred and twelve (112) case samples (101 postmortem case samples and 11 antemortem case samples) from drug-toxicity related death investigations completed by the toxicology laboratory of the Office of the Chief Medical Examiner, Edmonton, Alberta, Canada. Analytical data is presented which compares concentrations of ortho-methylfentanyl in paired postmortem blood collected from both a visualized, ligated femoral vein together with postmortem blood collected directly from the heart, ie, visualized. Median blood ortho-methylfentanyl concentrations were found to be 5.94 ng/mL (femoral) and 8.04 ng/mL (cardiac). The median cardiac to femoral blood concentration ratio across the entire data set was 1.19. The study highlights the varied distribution in the body based on the median concentration of these drugs in postmortem blood.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144225558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Thallium distribution along segmented single hairs in a case of a criminal poisoning.","authors":"Peter Heitland, Fritz Pragst, Sven Hartwig, Helmut D Köster","doi":"10.1093/jat/bkaf034","DOIUrl":"https://doi.org/10.1093/jat/bkaf034","url":null,"abstract":"<p><p>In this study, six single hairs, each measuring 9-11 cm in length, from a victim of a single criminal thallium (Tl) poisoning incident were analyzed in 0.3 cm segments applying a validated inductively coupled plasma mass spectrometry (ICP-MS) method. The results hold significant forensic value, particularly in cases involving limited sample material. Despite the very low weight of 0.3 cm single hair (12-16 µg) we found that the sensitivity of ICP-MS is sufficient to carry out the section-by-section analysis even in one single hair for determining Tl in poisoning cases. The measured Tl concentrations in this case were determined to be in the range of 0.6-6.5 µg/g hair, which are 100-fold beyond what is normally found in the German population. The consistent decrease in concentration from proximal to distal segments could be interpreted by predominant Tl incorporation via sweat. This finding is discussed in comparison with previous studies on Tl hair concentrations and in relation to the toxic effects of thallium on hair growth and sweat gland function. Due to the low limit of detection of 0.008 µg Tl/g hair, we conclude that Tl poisoning can be detected by analysis of single hairs in 0.3 cm segments. However, proving repeated or continuous exposure to the toxin may be challenging due to its incorporation from sweat.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144225559","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of Alexamorelin Consumption Biomarkers Using Human Hepatocyte Incubations and High-Resolution Mass Spectrometry.","authors":"Elizabeth Pobee, Gloria Daziani, Prince S Gameli, Giuseppe Basile, Jeremy Carlier, Anastasio Tini","doi":"10.1093/jat/bkaf038","DOIUrl":"https://doi.org/10.1093/jat/bkaf038","url":null,"abstract":"<p><p>Alexamorelin is a synthetic peptide and growth hormone secretagogue (GHS) with potential performance-enhancing properties, making its use and abuse a topic of interest in clinical research and doping monitoring. Alexamorelin mimics the natural peptide hormone ghrelin by binding to the GHS type 1a receptor (GHS-R1a) in the pituitary gland, thereby promoting endogenous growth hormone release. Identifying alexamorelin and/or its metabolite biomarkers is crucial for effective doping controls. The purpose of this study was to determine and characterize biomarkers associated with alexamorelin intake. In silico metabolite predictions were performed using GLORYx freeware, and in vitro incubations were conducted with pooled human hepatocytes from 10 donors. Samples were analyzed using liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS), with data processed through Thermo Scientific's Compound Discoverer. GLORYx predicted 21 single-reaction metabolites. N-Acetylation was identified as the primary transformation, with the highest probability score (98%), and occurring either at the C-terminal Ala or the N-terminal Lys. Other predicted transformations included N-oxidation, hydroxylation, amide hydrolysis, oxidative deamination, and phase II N-glucuronidation, with probability scores below 40%. All these transformations were predicted to occur at the two C-terminal (Ala or His) or N-terminal (D-Phe or Lys) amino acids. After 3 h of incubation with hepatocytes, only one metabolite (known as examorelin or hexarelin) was detected, resulting from the C-terminal cleavage of the Ala amino acid; this metabolic reaction is mediated by a carboxypeptidase. The alexamorelin signal decreased approximately 150-fold after 3 h, indicating significant hepatic metabolism. However, examorelin itself is a commercially available GHS secretagogue, and thus, it is not specific to alexamorelin consumption. Detecting alexamorelin remains critical to documenting its use.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144225557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simultaneous Screening and Quantitation of Drugs and their Metabolites in Post-Mortem Vitreous Humor by Liquid Chromatography-High Resolution Mass Spectrometry.","authors":"Edmund Rab, Ellen Sellers, Marta-Sofia Lindo-Cardoso, Gabrielle Wall, Faizan Khan","doi":"10.1093/jat/bkaf049","DOIUrl":"https://doi.org/10.1093/jat/bkaf049","url":null,"abstract":"<p><p>Post-mortem vitreous humor may be used for toxicological analysis if blood and urine are unavailable, or where post-mortem blood is thought to be affected by post-mortem changes. Use of vitreous humor has been restricted by the available sample volume and instrument sensitivity. However, the advent of combined screening and quantitative methodologies using liquid chromatography-high resolution mass spectrometry (LC-HRMS) makes analysis of vitreous humor possible. This study examines an existing combined screening and quantitative methodology to determine if it is suitable for use with vitreous humor. Analysis of standard solutions containing 48 compounds showed % difference between expected and measured values in the range -15.59 to 20.81, -15.73 to 18.34, -14.32 to 19.77 and -19.90 to 19.78 for very low, low, mid and high range standard solutions respectively. Intraassay %CV was in the range 0.93 to 10.10, 1.35 to 15.19, 3.07 to 11.56 and 2.04 to 8.29 and interassay %CV was 0.96 to 17.40, 3.68 to 17.03, 3.94 to 17.12 and 4.87 to 16.55. Limits of quantitation range from 0.002 to 0.5 and limits of detection from 0.0008 to 0.06 mg/L. There was no significant interference from ion suppression or isobaric compounds and very little carryover. Dilution 1:2, 1:5 and 1:10 with vitreous humor gave acceptable results. Comparison of screening results from 129 post-mortem cases showed that most compounds detected in blood and/or urine were also detected in vitreous humor. Compounds more readily detected in vitreous humor included 6-monoacetylmorphine, cocaine, codeine, dihydrocodeine, olanzapine, desmethylzopiclone, diazepam, cocaethylene and desmethylmirtazapine. Compounds more readily identified in blood and/or urine included desmethylsertraline, EDDP, nordiazepam, papaverine, paracetamol and morphine. The assay is suitable for screening and quantitation of drugs and their metabolites in vitreous humor and can be used where blood and urine are unavailable, or where the analysis of vitreous humor may provide useful information.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144208527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Polydrug fatal intoxication involving MDPHP: Detection and in silico investigation of multiple 3,4-methylenedioxy-derived designer drugs and their metabolites.","authors":"Sara Casati, Alessandro Ravelli, Michele Dei Cas, Roberta F Bergamaschi, Sofia Vanerio, Lea Sicuro, Chiara Faraone, Marta Rossi, Nicola Galante, Luca Mollica, Gabriella Roda, Paola Rota, Alessio Battistini","doi":"10.1093/jat/bkaf048","DOIUrl":"https://doi.org/10.1093/jat/bkaf048","url":null,"abstract":"<p><p>A drug-related fatality involving 3,4-methylenedioxy-α-pyrrolidinohexanophenone (MDPHP) is here reported. Belonging to the class of synthetic cathinones (SCs), MDPHP is a 3,4-methylenedioxy-derived designer (MDDs) drug with a pyrrolidine moiety and an alkyl portion with six carbon atoms. Other MDD pyrrolidine derivatives belong to the alkyl homologous series (C3-C5) and are known as 3,4-methylenedioxy-α-pyrrolidinopropiophenone (MDPPP), 3,4-methylenedioxy-α-pyrrolidinobutyrophenone (MDPBP) and 3,4-methylenedioxypyrovalerone (MDPV). MDDs are psychostimulant drugs of abuse that primarily act on monoamine transporters; little is known about their off-target liability. Recently, MDPHP has gained attention due to increasing seizures and involvement in human intoxications, but currently there is a lack of data about its pharmaco-toxicological effects. In the case reported here, a 58-year-old man with a history of MDPV addiction was found dead in a waterway. While no evidence of natural disease or trauma was found to account for the death, toxicological analysis revealed the presence of MDPHP in addition to MDPPP, MDPV, MDPBP, clonazepam and citalopram. Since no standards of MDPPP and MDPBP were available at the time of the analysis, LC-QTOF analysis of the drugs and their metabolites were performed. The following concentrations of MDPHP were reported: 354.5 ng/mL in femoral blood (FB), 110.9 ng/mL in cardiac blood (CB), 1900 ng/mL in urine, 3000 ng/mL in bile, 490 ng/g in kidney, 80 ng/g in liver, 480 ng/g in lung, 98 ng/g in brain, 700 ng/mL in gastric content and 8.3 ng/mg in pubic hair. Other MDDs concentrations in biological fluids and tissue were significantly lower than MDPHP suggesting their presence as synthetic impurities. Finally, to better understand the binding properties of the abovementioned MDDs to several documented transporters and receptors, an in-silico evaluation was performed. The medical examiner reported that the cause of death was an acute multidrug intoxication by MDPHP and clonazepam in presence of MDPPP, MDPV, MDPBP and citalopram.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144181844","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of Cannabinoids and Semi-synthetic Cannabinoids in Authentic Breastmilk by Liquid Chromatography-Tandem Mass Spectrometry.","authors":"Marco Ballotari, Michael T Truver, Nayana A Sojin, Rhea Parimoo, Lauren A Agliano, Jennifer L Hoyer, Amie J Goodin, Deepthi S Varma, Chris W Chronister, Kay Roussos-Ross, Bruce A Goldberger","doi":"10.1093/jat/bkaf047","DOIUrl":"https://doi.org/10.1093/jat/bkaf047","url":null,"abstract":"<p><p>Marijuana (cannabis) is generally considered the most frequently misused substance during pregnancy. The prevalence in the use of either medical or non-medical marijuana for relief of pregnancy-related symptoms is increasing, as well as the use of cannabis-related products containing cannabidiol (CBD) and semi-synthetic cannabinoids (SSCs). Δ9-tetrahydrocannabinol (THC) and CBD are highly lipophilic substances and will readily pass into breastmilk upon ingestion. The solubility of THC and CBD in lipids poses significant analytical challenges in extracting and identifying these substances in breastmilk. The aim of this study was to develop a new and sensitive assay utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the detection of cannabinoids in breastmilk. The method was optimized to quantitate Δ8-THC, Δ9-THC, cannabigerol (CBG), CBD, and cannabidiolic acid (CBDA) and validated with the guidance of the American Academy of Forensic Sciences Standards Board (ASB) Standard 036. The assay was then used to analyze breastmilk samples (N = 57) collected postpartum from female patients enrolled in a study assessing use behaviors of medical marijuana, non-medical marijuana, and CBD. All analytes passed validation criteria. Calibration curves for all analytes ranged 0.5-400 ng/mL, with the LOD and LLOQ of the method set at the lowest calibrator concentration. Δ9-THC was quantitated in 19 samples (33.3%) with a concentration range of 0.5-291 ng/mL. Δ8-THC was detected in one sample (1.8%) at 0.8 ng/mL, while CBD was observed in 3 samples at a concentration <LLOQ, and quantitated in only one sample (1.8%) also at a concentration of 0.8 ng/mL. CBG was detected in 7 samples (12.2%) with a concentration range of 0.6-12.9 ng/mL, and at a concentration <LLOQ in 12 samples. This study presents a sensitive method for the analysis of cannabinoids in breastmilk to support the follow-up assessments of marijuana and CBD use during pregnancy and postpartum.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144142654","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and Validation of an Analytical Method for the Determination of Select 4-Position Ring-Substituted Tryptamines in Plasma by Liquid Chromatography-Tandem Mass Spectrometry.","authors":"Ana Miguel Fonseca Pego, Malik Schoffner, Vamshikrishna Reddy Sammeta, Marilyn Naeem, David R Manke, Andrew Chadeayne, Grant C Glatfelter, Michael H Baumann, Marta Concheiro","doi":"10.1093/jat/bkaf045","DOIUrl":"https://doi.org/10.1093/jat/bkaf045","url":null,"abstract":"<p><p>4-Phosporyloxy-N, N-dimethyltryptamine (psilocybin) is a psychedelic tryptamine found in certain mushroom species that has shown efficacy in the treatment of various psychiatric disorders. In conjunction with the renewed interest in therapeutic effects of psychedelics, there has been an increase in psilocybin-like designer tryptamines appearing in non-medical drug markets. The present study aimed to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for detecting and quantifying 4-position ring-substituted tryptamines and their 4-hydroxy metabolites in plasma. Specifically, we investigated 4-phosphoryloxy-N, N-dimethyltryptamine (psilocybin), 4-acetoxy-N, N-dimethyltryptamine (psilacetin), 4-propionoxy-N, N-dimethyltryptamine (4-Pro-DMT) and their shared metabolite 4-hydroxy-N, N-dimethyltryptamine (psilocin), along with 4-methyl carbonato-N, N-di-n-propyltryptamine (4-MeCO3-DPT) and its metabolite 4-hydroxy-N, N-di-n-propyltryptamine (4-HO-DPT). Mass spectrometry analysis employed electrospray ionization (ESI) in positive mode, with two multiple reaction monitoring (MRM) transitions per analyte. Plasma samples were acidified with ascorbic acid, followed by protein precipitation with acetonitrile. Linearity was achieved across a concentration range of 0.5-100 ng/mL for all analytes, except psilocybin, which displayed linearity from 5-100 ng/mL. Validation results demonstrated acceptable bias (±20%) and imprecision (<20%) for all analytes. Matrix effects, evaluated in 10 samples (CV < 18.3%), indicated minimal interference, although ion enhancement was observed for psilocin (31.9%) and psilocybin (45.7%). Extraction efficiency across all tryptamines was approximately 50%. The assay method was used to quantitate plasma samples from male rats treated with 1.0 mg/kg s.c. of the prodrug psilacetin, and collected before and 5, 30, 60, 120 and 240 min after injection. No psilacetin was detected, and psilocin concentrations ranged from non-detected up to 32.7 ng/mL. Overall, we successfully developed a sensitive and specific method for the detection and quantification of six tryptamines in plasma, providing a robust tool for future research and clinical applications.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144142656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}