Journal of analytical toxicology最新文献

{"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}

{"title":"A novel screening workflow for nitazene analogs using LC-MS/MS precursor ion scan acquisition.","authors":"Amanda L Pacana, Britni N Skillman","doi":"10.1093/jat/bkaf046","DOIUrl":"https://doi.org/10.1093/jat/bkaf046","url":null,"abstract":"<p><p>A persistent problem in the detection of novel psychoactive substances (NPS) is the inability of traditional screening methodologies to rapidly adapt to evolving drug trends. As such, high-resolution mass spectrometry (HRMS) screening methods have gained popularity in recent years for the ability to use non-targeted acquisition to detect a wide variety of compounds without necessarily returning to method development. However, these instruments may be unattainable for some forensic laboratories due to the associated high capital costs. The described method provides an alternative screening method using precursor ion scan (PIS) acquisition on a liquid chromatography tandem mass spectrometry (LC-MS/MS) platform to screen for nitazene analogs. Four ions were evaluated (m/z 72.1, 98.0, 100.1, and 112.1) for D0 analytes and one ion (m/z 104.1) for the metodesnitazene-D4 internal standard. Using a liquid-liquid extraction in whole blood, the method was validated with a 0.5 ng/mL limit of detection and 1.0 ng/mL administrative cutoff. Observed matrix effects did not affect limit of detection and there was no demonstration of carryover or interferences. As a proof-of-concept study, authentic (n = 3) and blind fortified (n = 20) samples were evaluated using this method, which was able to identify all nitazenes with no false negatives or positives. Several nitazenes not initially included in the scope of method development or validation were also presumptively identified. To accommodate this novel instrumental analysis, a workflow is also proposed to assist in the identification of known and emerging nitazene analogs. LC-MS/MS is widely available among forensic laboratories and presents a viable alternative to HRMS screening for nitazene analogs when operated in PIS acquisition, in such cases that HRMS is unavailable for assessing emerging NPS threats.</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":"144142652","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":"Toxicology Testing in the United States: What the 2018 Census of Medical Examiner and Coroner Offices Tells Us.","authors":"Hope Smiley-McDonald, Sean Wire, Nichole D Bynum, Katherine M Bollinger, Kelly A Keyes, Jeri D Ropero-Miller","doi":"10.1093/jat/bkaf044","DOIUrl":"https://doi.org/10.1093/jat/bkaf044","url":null,"abstract":"<p><p>In 2021, the U.S. Bureau of Justice Statistics (BJS) published results for the 2018 Census of Medical Examiner and Coroner Offices (CMEC) that provided an update on the medicolegal death investigation system in the U.S. The 2018 Census collected data regarding toxicology service provisions, staffing, infrastructure, and practices, some of which were not included in the 2021 published BJS report from more than 1,600 responding medical examiner/coroner offices (MECs). The 2018 CMEC was conducted from June 2019 through March 2020 by mail, online, and email. Toxicology-related CMEC data were obtained from BJS's publicly accessible dataset and evaluated in this study. Results from this study include information on toxicology service capability across MECs, including the number and salary of forensic toxicologists, toxicology retention time schedules, laboratory accreditation, professional certification, drug screening practices at the death scene, and whether they request confirmation testing. Overall, internal capabilities for toxicology testing were rare in 2018, with only 78 MECs (5.9%) reporting this function. Large MECs, serving a population of 250,000 or more, comprised about 15% of MECs that responded to the toxicology testing questions, with the rest being evenly divided between MECs that serve small (<25,000) and medium sized (25,000-249,999) populations. Overall, 57.4% (n = 761) of MECs indicated that their forensic toxicology testing strategy has changed because of the increase in drug-related deaths, 53.9% of MECs (n = 715) perform drug screening tests, and 95.1% (n = 674) confirmed these tests with laboratory toxicology testing. Less than half of MECs reported that they had a toxicology specimen retention schedule (45.3%) or a computerized case management system (44.8%). These data are key to understanding (a) postmortem toxicology policies and practices, (b) how these practices have evolved, (c) MEC infrastructure; and (d) the national importance of these data considering the ongoing drug overdose crisis.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144132344","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":"Comprehensive LC-MS-MS analysis of THC isomers, analogs, homologs, and metabolites in blood and urine.","authors":"Luette S Muir, Sarah E Doumit, Joshua Z Seither, Jessica L Knittel, Jeffrey P Walterscheid, Erin L Karschner","doi":"10.1093/jat/bkaf023","DOIUrl":"10.1093/jat/bkaf023","url":null,"abstract":"<p><p>The legalization of hemp and the commercialization of hemp-based extracts have resulted in numerous cannabinoids appearing in consumer products. Although human pharmacological data are lacking for many of these isomers, analogs, and homologs of delta-9-tetrahydrocannabinol (Δ9-THC), these cannabinoids may be capable of inducing cannabimimetic effects. Structural similarities also pose unique analytical challenges due to overlapping retention times and ion transitions used to distinguish between various parent drugs and metabolites. Therefore, traditional cannabinoid assays containing Δ9-THC, 11-hydroxy-Δ9-THC (11-OH-Δ9-THC), and 11-nor-9-carboxy-Δ9-THC (Δ9-THCCOOH) are no longer sufficient to confront this new threat to public safety. A new method has been developed and validated to quantitatively confirm Δ8- and Δ9-THC, their hydroxylated and carboxylated metabolites, and 9(R)- and 9(S)-hexahydrocannabinol (HHC) stereoisomers in blood and qualitatively identify these analytes in urine. This method is also capable of qualitatively confirming Δ9,11-THC (exo-THC), HHCCOOH, Δ6a,10a-THCCOOH/Δ10-THCCOOH, and Δ8 and Δ9 THC homologs including tetrahydrocannabivarin (THCV), tetrahydrocannabutol (THCB), tetrahydrocannabihexol (THCH; Δ8-THCH in urine only), tetrahydrocannabiphorol (THCP), THC-C8, as well as 9(R)- and 9(S)-hexahydrocannabiphorol (HHCP) in blood and urine. Limits of detection were 1 ng/mL for non-carboxylated analytes and 5 ng/mL for carboxylated analytes. Calibration curves for parent and hydroxylated THC isomers and HHC stereoisomers were 1 to 50 ng/mL, whereas calibration curves for the carboxylated THC isomers were 5-250 ng/mL. This method separates all analytes of interest from potential synthesis byproducts such as Δ8-iso-THC, Δ4(8)-iso-THC, and exo-THC. Unambiguous identification of these cannabinoids will increase forensic toxicology reporting accuracy while navigating the changing landscape of cannabis regulation and product formulation.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":"322-331"},"PeriodicalIF":2.3,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143648775","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":"Quantification of phosphatidylethanol 16:0/18:1 in blood using supercritical fluid chromatography-tandem mass spectrometry.","authors":"Munchelou M Gomonit, Markus Roman, Britni N Skillman, Michael T Truver, Robert Kronstrand","doi":"10.1093/jat/bkaf007","DOIUrl":"10.1093/jat/bkaf007","url":null,"abstract":"<p><p>Phosphatidylethanol (PEth) consists of phospholipids synthesized in erythrocyte cell membranes in the presence of ethanol and serves as a sensitive and specific indicator of alcohol consumption. Further research on PEth formation, degradation, and stability in postmortem (PM) samples would support its routine application in forensic toxicology. A supercritical fluid chromatography-tandem mass spectrometry (SFC-MS-MS) method was developed and validated to quantify PEth 16:0/18:1 in blood. PEth 16:0/18:1 was extracted from blood (0.25 g) using an 8:2 (v/v) heptane:2-propanol mixture. Method validation results met American National Standards Institute/Academy Standards Board 036 guidelines. Recovery was >48%, and matrix effects were <20%. The linear range was 10-2500 ng/g, and lower limit of quantification was 10 ng/g. Bias was ±17.7%, and precision was <17.1% for all quality control levels. Carryover, endogenous, and exogenous interferences were negligible. Extracts were stable beyond 72 hours. In a proof-of-concept study reanalyzing 35 PM case samples, PEth concentrations ranged between 32.6 to 2476 ng/g. Short-term stability studies showed that fortified bovine blood (200 ng/g) preserved with 0.4% sodium fluoride (NaF) stored at room temperature had a 6.6% concentration drop after 48 hours, while blood stored at 4°C decreased by 13.5% over 14 days. Additionally, human PEth-positive blood preserved with 0.4% NaF showed a 6.7% decrease in in vivo PEth concentrations compared to a 17.5% decrease in heparin-preserved blood after 14 days at 4°C, supporting the use of 0.4% NaF in reducing PEth degradation over time. An in vitro model was also developed to simulate early PM PEth changes. Results found that PEth formation occurred in an ethanol concentration-dependent manner with minimal degradation, and considerations should be taken when interpreting PEth concentrations in cases with long PM interval, and if the decedent had a high blood alcohol concentration level and was left at elevated temperatures. This is the first SFC-MS-MS method successfully developed and validated for the analysis of PEth in PM samples.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":"289-298"},"PeriodicalIF":2.3,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143624752","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":"A comparison of vitreous fluid and blood matrices in postmortem drug analysis.","authors":"Erin B Divito, Jedediah I Bondy, Zachary J DiPerna, Frederick W Fochtman, Christopher B Divito","doi":"10.1093/jat/bkaf015","DOIUrl":"10.1093/jat/bkaf015","url":null,"abstract":"<p><p>Peripheral blood is considered the gold standard for postmortem toxicological analysis. However, vitreous fluid (VIT) has been described as more resistant to postmortem redistribution and may act as an isolated matrix, preserving postmortem drug concentrations. To determine the utility of VIT analysis compared to traditional blood analysis for 6-acetylmorphine, morphine, cocaine, benzoylecgonine, fentanyl, and norfentanyl, paired peripheral blood and VIT were collected and analyzed in 122 postmortem cases from Western Pennsylvania. In this study, we tested the frequency of detection and performed analyses to correlate drug concentrations in both matrices. We demonstrate that VIT provides a viable matrix for the postmortem analysis of several drugs of abuse and their metabolites. However, when both matrices are available for analysis, VIT, when compared to whole blood, is not optimal for all analytes. These differences are likely due to differences in physiochemical properties and other pharmacokinetic parameters.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":"351-357"},"PeriodicalIF":2.3,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143556955","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":"Green liquid-liquid microextraction for quantification of ketamine and metabolites in human urine by gas chromatography-mass spectrometry.","authors":"Hsueh-Hui Yang, Chia-Sui Kao, Ahai C Lua, Tsong-Yung Chou","doi":"10.1093/jat/bkaf022","DOIUrl":"10.1093/jat/bkaf022","url":null,"abstract":"<p><p>This study aimed to develop and validate a green method for the detection of ketamine and its metabolites in human urine samples and subsequently determine which metabolite is more suitable for judgment of ketamine abuse. Ketamine and its metabolites were extracted from urine samples with 1-undecanol using liquid-liquid microextraction based on the solidification of floating organic droplet extraction. The extracts were directly analyzed using gas chromatography-mass spectrometry (GC-MS) without derivatization. The sample pretreatment procedure prior to GC-MS analysis was simple, fast, and required little solvent consumption. Parameters that affect the extraction efficiency, including pH of the urine sample and centrifugation speed, were optimized. Under the optimized conditions, the limits of detection for norketamine, dehydronorketamine, and ketamine were 1.5, 1.7, and 1.0 ng/mL, respectively. The method was used to analyze eight real urine samples from drug abusers and exhibited acceptable accuracy and precision. By analyzing real samples, this study revealed that detection of dehydronorketamine in urine samples for the judgment of ketamine abuse may be more suitable than detection of norketamine or ketamine. The method used in this study addresses the need for green analytical techniques in toxicology.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":"315-321"},"PeriodicalIF":2.3,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143605041","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 4F-MDMB-BICA using a molecular network strategy in a case of severe poisoning with coma.","authors":"Weniko Caré, Romain Magny, Dominique Vodovar, Frédérik Bélot-de Saint-Léger, Jérôme Langrand, Hervé Laborde-Castérot, Laurence Labat, Pascal Houzé","doi":"10.1093/jat/bkaf019","DOIUrl":"10.1093/jat/bkaf019","url":null,"abstract":"<p><p>Synthetic cannabinoids remain one of the most important groups of new psychoactive substances and are responsible for many cases of poisoning in Europe. Deaths from acute 4F-MDMB-BICA poisoning have recently been reported. Severe poisonings may be underreported because 4F-MDMB-BICA is not routinely screened for in most forensic and toxicology laboratories. We report the case of a young man in France who presented with poisoning after orally consuming a powdered substance sold online as an opioid. The coma required intensive care unit management with emergent chest tube insertion and mechanical ventilation. The outcome was favorable with no sequelae due to early medical care. In the absence of remaining product and preserved urine samples, qualitative toxicological screening was performed on plasma, cerebrospinal fluid, and a hair strand. Using ultra-high-performance liquid chromatography-high-resolution tandem mass spectrometry and a molecular network data processing strategy, 4F-MDMB-BICA and two of its metabolites were identified only in plasma and cerebrospinal samples. These results were consistent with a single exposure. The identification of the substance consumed was crucial because of discrepancy between the symptoms observed and those expected after presumed exposure. Identification of 4F-MDMB-BICA and two of its metabolites was achieved in early plasma and cerebrospinal fluid samples. This documented case is helping to improve knowledge of 4F-MDMB-BICA poisoning, which could be an emerging public health issue.</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":"364-368"},"PeriodicalIF":2.3,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143556873","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":"HIV combination drug therapies: development and validation of an LC-MS-MS method for simultaneous quantitation of abacavir, dolutegravir, and lamivudine in rat matrices in support of toxicology studies.","authors":"Melanie A Rehder Silinski, Jennifer A Gilliam, Julia Apoian, Brenda L Fletcher, Reshan A Fernando, Suramya Waidyanatha","doi":"10.1093/jat/bkaf016","DOIUrl":"10.1093/jat/bkaf016","url":null,"abstract":"<p><p>Abacavir (ABC), Dolutegravir (DTG), and Lamivudine (3TC) are part of a fixed-dose combination medication for the treatment of HIV. The three drugs offer different but complementary mechanisms of action by inhibiting reverse transcriptase and integrase, and ultimately inhibiting HIV replication. Due to the lack of information regarding long-term safety following in utero exposure, we are evaluating potential toxicity to offspring following in utero exposure to this combination therapy in Hsd:Sprague Dawley®SD® (HSD) rats, including cardiovascular toxicity and neurotoxicity. Generating internal exposure data are integral to putting toxicological findings into context. The objective of this work was to develop and validate a method to simultaneously quantitate ABC, DTG, and 3TC in rat matrices following exposure to this combination. The method used protein precipitation of plasma, fetal, placental, brain, or heart homogenate, followed by ultra-performance liquid chromatography-tandem mass spectrometry. In adult Sprague Dawley rat plasma, the method was linear (r ≥ 0.99) over the range 10/15/5 to 10,000/15,000/5000 ng/mL for ABC/DTG/3TC and recovery was ≥92% for all three analytes at all concentration levels. The limits of detection were 2.22, 3.69, and 0.978 ng/mL for ABC, DTG, and 3TC, respectively. Intra- and inter-day precision was ≤8.7% relative standard deviation (RSD), and relative error (RE) ≤±12.0% for standards prepared at 20/30/10, 400/600/200, and 5000/7500/2500 ng/mL. Matrix standards as high as 40/60/20 µg/mL could be diluted into the calibration range (RE≤±3.5% and RSD ≤2.4%). The method was evaluated for HSD rat maternal plasma and fetal, placental, brain, and heart homogenates (mean RE ≤±15.0% and RSD ≤8.6%). Analyte stability was demonstrated in extracted plasma for 2 days at different temperatures, and in various matrices stored at -80°C for at least 32 days (80-113% of Day 0 concentrations). These data demonstrate that this simple and efficient method is suitable for quantitation of ABC, DTG, and 3TC in rat matrices generated from toxicology studies. The method can easily be adapted to other biological matrices and species (e.g. human).</p>","PeriodicalId":14905,"journal":{"name":"Journal of analytical toxicology","volume":" ","pages":"332-339"},"PeriodicalIF":2.3,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143556872","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}