Journal Of Analytical Toxicology[JOURNAL]
Sun
200 papers
Comparison of a comprehensive liquid chromatography-quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS) screen in whole blood with conventional immunoassay-based techniques.
Ayala J, Kerrigan S
J Anal Toxicol · 2025 Nov · PMID 40498580 · Publisher ↗
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 emerg...
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 and 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.
Changes in blood cannabinoid concentrations over multiple collection times in driving under the influence of drugs casework.
Peterson BL, Hessler MR
J Anal Toxicol · 2025 Oct · PMID 40478690 · Publisher ↗
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...
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 the 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 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 to 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 to 16 ng/mL. THC-COOH concentrations (n = 81) ranged from 7.1 to 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.
Identification of alexamorelin consumption biomarkers using human hepatocyte incubations and high-resolution mass spectrometry.
Pobee E, Daziani G, Gameli PS … +3 more , Basile G, Carlier J, Tini A
J Anal Toxicol · 2025 Jul · PMID 40465419 · Publisher ↗
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. Alexamor...
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 analysed 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.
Thallium distribution along segmented single hairs in a case of a criminal poisoning.
Heitland P, Pragst F, Hartwig S … +1 more , Köster HD
J Anal Toxicol · 2025 Sep · PMID 40465418 · Publisher ↗
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...
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 materials. 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 the 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.
LC-MS/MS determination of the novel fentanyl analog, ortho-methylfentanyl, in drug-related toxicity casework: concentrations in ligated femoral blood.
Chatterton CN, Handy RP, Shoemaker GK
J Anal Toxicol · 2025 Nov · PMID 40465412 · Publisher ↗
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 prot...
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 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, that is, 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.
Simultaneous screening and quantitation of drugs and their metabolites in postmortem vitreous humor by liquid chromatography-high resolution mass spectrometry.
Rab E, Sellers E, Lindo-Cardoso MS … +2 more , Wall G, Khan F
J Anal Toxicol · 2025 Nov · PMID 40459883 · Publisher ↗
Postmortem vitreous humor may be used for toxicological analysis if blood and urine are unavailable or where postmortem blood is thought to be affected by postmortem changes. Use of vitreous humor has been restricted by...
Postmortem vitreous humor may be used for toxicological analysis if blood and urine are unavailable or where postmortem blood is thought to be affected by postmortem 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 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 postmortem 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.
Polydrug fatal intoxication involving MDPHP: Detection and in silico investigation of multiple 3,4-methylenedioxy-derived designer drugs and their metabolites.
Casati S, Ravelli A, Dei Cas M … +10 more , Bergamaschi RF, Vanerio S, Sicuro L, Faraone C, Rossi M, Galante N, Mollica L, Roda G, Rota P, Battistini A
J Anal Toxicol · 2025 Jul · PMID 40440133 · Full text
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 w...
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: 350 ng/mL in femoral blood (FB), 110 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 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.
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.
Pego AMF, Schoffner M, Sammeta VR … +6 more , Naeem M, Manke DR, Chadeayne A, Glatfelter GC, Baumann MH, Concheiro-Guisán M
J Anal Toxicol · 2025 Jul · PMID 40418247 · Full text
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...
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 to 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.
Analysis of cannabinoids and semi-synthetic cannabinoids in authentic breastmilk samples by liquid chromatography-tandem mass spectrometry.
Ballotari M, Truver MT, Sojin NA … +8 more , Parimoo R, Agliano LA, Hoyer JL, Goodin AJ, Varma DS, Chronister CW, Roussos-Ross K, Goldberger BA
J Anal Toxicol · 2025 Oct · PMID 40418239 · Publisher ↗
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 increa...
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.
A novel screening workflow for nitazene analogs using LC-MS/MS precursor ion scan acquisition.
Pacana AL, Skillman BN
J Anal Toxicol · 2025 Oct · PMID 40418229 · Publisher ↗
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 (H...
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.
Toxicology testing in the USA: what the 2018 census of medical examiner and coroner offices tells us.
Smiley-McDonald H, Wire S, Bynum ND … +3 more , Bollinger KM, Keyes KA, Ropero-Miller JD
J Anal Toxicol · 2025 Oct · PMID 40408152 · Publisher ↗
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 USA....
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 USA. 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 1600 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 (i) postmortem toxicology policies and practices, (ii) how these practices have evolved, (iii) MEC infrastructure, and (iv) the national importance of these data considering the ongoing drug overdose crisis.
Comparison of Δ9-tetrahydrocannabinol in venous and capillary blood following ad libitum cannabis smoking by occasional and daily users.
Dooley G, Godbole S, Wrobel J … +4 more , Henthorn T, Brooks-Russell A, Limbacher S, Kosnett M
J Anal Toxicol · 2025 Sep · PMID 40366742 · Full text
Δ9-Tetrahydrocannabinol (Δ9-THC) is the most prominent and main psychoactive cannabinoid found in cannabis. In forensic matters involving cannabis, such as drugged driving or workplace accident investigations, blood Δ9-T...
Δ9-Tetrahydrocannabinol (Δ9-THC) is the most prominent and main psychoactive cannabinoid found in cannabis. In forensic matters involving cannabis, such as drugged driving or workplace accident investigations, blood Δ9-THC determination is typically required. Venipuncture by a phlebotomist at a medical facility is often the standard blood collection protocol, but this procedure is time consuming and requires specialized training. Capillary blood collection at the site of a transportation or workplace mishap may provide a collection method that is logistically easier and may better reflect blood cannabinoid concentrations at the time of an incident. This study represents the first temporal comparison of the concentration of Δ9-THC and its primary metabolites in venous and capillary blood obtained from users following ad libitum inhalation of contemporary high-concentration cannabis products. Participants provided their own cannabis from a licensed Colorado dispensary and were instructed to smoke or vape ad libitum the amount most used for the desired effect during a 15-minute period. Capillary blood samples collected at the lateral shoulder using the TAP® II microneedle device and standard venipuncture samples at the forearm were collected contemporaneously at baseline and then 10, 30, 60, 90, and 140 minutes after the last inhalation and were analyzed for Δ9-THC, 11-hydroxy-Δ9-THC, and 11-carboxy-Δ9-THC by liquid chromatography-tandem mass spectrometry. Within-subject Δ9-THC concentrations trended lower, often up to 30 to 40%, in contemporaneous capillary blood samples than in venous blood samples until 140 min after cannabis smoking. Concentrations of the Δ9-THC metabolites 11-hydroxy-Δ9-THC and 11-carboxy-Δ9-THC were equivalent at all but the first timepoint after smoking. Due to logistical advantages, capillary blood collection by microneedle devices may be a viable option for qualitative detection of Δ9-THC and its metabolites soon after an incident or a quantitative determination if the samples are collected at least 2 hours after cannabis inhalation.
Evaluation of DrugWipe® 6S with the WipeAlyser® reader for drug screening of drivers.
Jamt REG, Gjerde H, Clausen GB … +2 more , Bache-Andreassen L, Øiestad EL
J Anal Toxicol · 2025 Sep · PMID 40354046 · Full text
On-site drug screening of oral fluid samples has gained attention because of its convenience and rapid results. The aim of this investigation was to compare the results of preliminary screening for drugs in oral fluid sa...
On-site drug screening of oral fluid samples has gained attention because of its convenience and rapid results. The aim of this investigation was to compare the results of preliminary screening for drugs in oral fluid samples collected from suspected drug-impaired drivers using DrugWipe 6S and WipeAlyser reader with the results obtained from blood samples. Additionally, we compared the DrugWipe test results with findings of drug traces detected within the used DrugWipe devices. Police officers selected a sample of 355 suspected drug-impaired drivers in 2023. They used DrugWipe 6S for preliminary drug screening of drivers. After the field drug testing of oral fluid, the apprehended drivers were brought to a physician for the collection of blood samples. The collected samples (DrugWipe devices and blood samples) were submitted to the Norwegian National Forensic Toxicology Laboratory for analysis. The proportion of positive DrugWipe results that were unconfirmed when analysing blood samples was 82% for opiates, 75% for cocaine, and ∼19%-20% for amphetamines, cannabis, and benzodiazepines. The proportion of negative DrugWipe results that were found positive in blood samples was for cannabis and benzodiazepines ∼13%-14%, and for other drugs <3%. Detected drug traces in the used DrugWipe devices corresponded well with DrugWipe readouts for cannabis, amphetamines, and cocaine. The lack of correspondence between DrugWipe test results for cocaine and findings in blood may be due to the fact that the concentration of cocaine in saliva is often much higher than in blood, and the DrugWipe test is very sensitive. In addition, degradation and elimination of cocaine before the blood sample is taken may contribute to cocaine concentrations below the cut-off concentration in blood. For opiates and benzodiazepines, traces of drugs were found in relatively few DrugWipe devices. Many unconfirmed positives for opiates were most likely due to cross-reaction with substances in 'snus' (snuff tobacco).
Protonitazene metabolite variability in postmortem casework.
Wood R, Moore R
J Anal Toxicol · 2025 Sep · PMID 40353845 · Publisher ↗
Protonitazene is a highly potent synthetic opioid that has recently emerged in the illicit drug supply. Data on its metabolism and the detection of its metabolites in postmortem specimens are limited. This study retrospe...
Protonitazene is a highly potent synthetic opioid that has recently emerged in the illicit drug supply. Data on its metabolism and the detection of its metabolites in postmortem specimens are limited. This study retrospectively analyzed 14 postmortem cases to investigate metabolite profiles in blood and urine using high-resolution mass spectrometry. Detected metabolites included 5-aminoprotonitazene, N-desethylprotonitazene, and 4-hydroxynitazene. Additionally, a novel metabolite, hydroxyprotonitazene, previously only observed in liver microsome models, was identified. Significant variability in the metabolites detected across cases was observed, likely influenced by differences in metabolism, postmortem changes, sampling methods, and metabolite stability. This study underscores the challenges of detecting protonitazene exposure and highlights the necessity of targeting multiple metabolites, as no single marker reliably indicated protonitazene use. Further research is needed to confirm metabolite identities, evaluate their stability, and understand their role in protonitazene toxicity.
Drug transfer during intimate moments can produce an adverse analytical finding during a doping control: A case report with ligandrol.
Kintz P, Gheddar L
J Anal Toxicol · 2025 Oct · PMID 40338642 · Publisher ↗
Selective androgen receptor modulators (SARMs) are a new class of substances that have similar properties to anabolic steroid agents, but with marked reduced androgenic properties. As SARMs have the potential to be misus...
Selective androgen receptor modulators (SARMs) are a new class of substances that have similar properties to anabolic steroid agents, but with marked reduced androgenic properties. As SARMs have the potential to be misused for performance enhancement in sport due to their anabolic properties as well as their ability to stimulate androgen receptors in the muscle and the bone, they have been prohibited at-all-times by the World Anti-Doping Agency (WADA) since 2008 under section S1.2 of the List. Ligandrol is one of the more popular SARMs. A WADA-accredited laboratory identified bishydroxy-ligandrol in the urine of a female athlete, the major ligandrol metabolite at approximately 90 pg/mL (specimen A) and 200 pg/mL (specimen B). The athlete challenged this anti-doping rule violation and requested a hair test to document possible incidental exposure. About 7 weeks after urine collection, a hair specimen (brown in color and > 20 cm in length) was collected and segmented in 6 × 1 cm segments. Ligandrol was tested by liquid chromatography-tandem mass spectrometry after alkaline incubation and extraction. With a limit of quantitation at 1 pg/mg, no ligandrol was identified. It appears that the athlete was unaware her husband was taking the substance, which was confirmed by his hair test (ligandrol at 7 and 8 pg/mg in 2 × 2.5 cm segments). The Court of Arbitration for Sports accepted the athlete's explanation that she had been exposed to ligandrol through the exchange of bodily fluids with her husband and lifted her provisional ban. This case demonstrates that drug transfer between two subjects is possible during intimate moments.
Medetomidine quantitation and enantiomer differentiation in biological specimens collected after fatal and non-fatal opioid overdoses.
Walton SE, Stang BN, Kacinko S … +3 more , Papsun DM, Logan BK, Krotulski AJ
J Anal Toxicol · 2025 Oct · PMID 40338638 · Publisher ↗
Medetomidine is an alpha-2 agonist and non-opioid sedative. Its presence in illicit fentanyl and heroin drug supplies poses significant risks to user health caused by cardiac effects and sedation. Medetomidine exists in...
Medetomidine is an alpha-2 agonist and non-opioid sedative. Its presence in illicit fentanyl and heroin drug supplies poses significant risks to user health caused by cardiac effects and sedation. Medetomidine exists in two enantiomeric forms: dexmedetomidine and levomedetomidine. Dexmedetomidine is used in humans in medical settings, while dexmedetomidine alone and a racemic mixture of dexmedetomidine and levomedetomidine are used in animals in veterinary settings. Little information is known about circulating blood concentrations of medetomidine or its enantiomers in humans in situations involving synthetic opioids (e.g. fentanyl) and other sedatives (e.g. xylazine, benzodiazepines). A new toxicological workflow using liquid chromatography-tandem quadrupole mass spectrometry (LC-QQQ-MS) was developed and validated for the quantitation of medetomidine and the qualitative enantiomeric separation of dexmedetomidine and levomedetomidine. The assays were applied to a case series of 100 authentic specimens from emergency department admissions and forensic postmortem investigations containing medetomidine, fentanyl, xylazine, and metabolites, among other substances. Medetomidine blood concentrations in non-fatal overdoses ranged 0.1-16 ng/mL (median: 1.5 ng/mL) and in fatal overdoses ranged 0.1-32 ng/mL (median: 0.31 ng/mL). Xylazine was co-detected in 76% of cases with higher median concentrations: 8.3 ng/mL (non-fatal) and 15 ng/mL (fatal). Fentanyl was co-detected in 93% of cases with median concentrations of 5.2 ng/mL (non-fatal) and 21 ng/mL (fatal). Dexmedetomidine and levomedetomidine were identified in 90% of cases; the remaining cases were confirmed or suspected medical-setting administration of dexmedetomidine. These findings demonstrate that medetomidine is arising from veterinary or clandestine sources, and we hypothesize the latter. Recreational medetomidine use causes adverse effects such as profound bradycardia and heightened sedation, especially when combined with fentanyl and xylazine. Forensic laboratories must remain aware of adulterants, like medetomidine, appearing in traditional opioid products (e.g. fentanyl, heroin), updating testing methods to capture these emerging adulterants in real-time.
A validated screening and confirmation method for 946 drugs and metabolites using LC-QTOF-MS with SWATH acquisition.
Sarkisian M, Rodda LN
J Anal Toxicol · 2025 Jul · PMID 40324907 · Publisher ↗
A streamlined liquid chromatography quadrupole time-of-flight mass spectrometry method utilizing protein precipitation and filtration extraction was developed to achieve rapid and reliable screening and confirmation for...
A streamlined liquid chromatography quadrupole time-of-flight mass spectrometry method utilizing protein precipitation and filtration extraction was developed to achieve rapid and reliable screening and confirmation for blood and urine matrices. This method targets 946 drugs and metabolites across 35 drug classes via sequential window acquisition of all theoretical mass spectra with variable customized windows to enhance spectral clarity, and was validated per established guidelines to ensure high accuracy and reproducibility. Combined with complementary in-house methods, this approach meets and exceeds the testing requirements outlined in ANSI/ASB standards and recommendations for postmortem, drug-facilitated crime, and Tier I and II driving under the influence of drug analyses. The method demonstrated efficient and sensitive performance, achieving limits of detection as low as 0.1 ng/mL. It accurately identified expected detections across 67 proficiency test samples and 224 authentic case samples, with high accuracy and reliability in the detection of both traditional drugs and novel psychoactive substances. The method employs an in-house built library and incorporates in-batch standards analyzed alongside case samples to ensure contemporaneous identification criteria, making it suitable for confirmation and reporting purposes. By expanding the analytical capabilities to include a vast range of analytes, this method improves the likelihood of identifying substances that may otherwise go undetected and reduces the need for multiple separate tests, thereby enhancing the overall effectiveness of toxicological investigations.
Ethanol production in the gut: an autopsy case.
Kusano M, Kohda C, Fujishiro M … +1 more , Matsuyama TA
J Anal Toxicol · 2025 Jul · PMID 40324906 · Publisher ↗
Postmortem alcohol production by microorganisms has been known to increase blood alcohol concentration, potentially leading to erroneous interpretation. Here, we present a peculiar case where postmortem toxicology detect...
Postmortem alcohol production by microorganisms has been known to increase blood alcohol concentration, potentially leading to erroneous interpretation. Here, we present a peculiar case where postmortem toxicology detected a high ethanol concentration only in the stomach content. An 87-year-old bedridden woman was taken to senior day care facility, where the nurse attempted to take her vitals but noticed she was not breathing. The facility called for an ambulance, but she was pronounced dead soon after arriving at the hospital. She was found to be severely dehydrated, and her blood tests indicated poor health. Autopsy was performed 2 days after death, and postmortem alcohol analysis detected a high ethanol concentration only in the stomach content. Our aim was to investigate the causative agent of ethanol production; microbiological analysis identified the yeast species Candida glabrata as the responsible microorganism.
Lateral flow immunoassay for amatoxins detection in human urine compared to liquid chromatography-high-resolution tandem mass spectrometry.
Vollmer AC, Bambauer TP, Bever CB … +3 more , Tam CC, Wagmann L, Meyer MR
J Anal Toxicol · 2025 Sep · PMID 40314120 · Publisher ↗
Amatoxin-containing mushrooms, which contribute to many intoxications each year, are of particular interest for clinicians and toxicologists, as patients require special treatment in hospitals. To confirm the presence of...
Amatoxin-containing mushrooms, which contribute to many intoxications each year, are of particular interest for clinicians and toxicologists, as patients require special treatment in hospitals. To confirm the presence of amatoxins, approaches for their fast, sensitive, and reliable identification must be available. Solid-phase extraction followed by liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS) is widely applied for analysis of amatoxins as this combination provides suitable sensitivity, specificity, and mass accuracy. Nevertheless, time-consuming preparatory steps and expensive equipment are required. Therefore, a lateral flow immunoassay (LFIA) for the trace detection of α-, β-, and γ-amanitin was established and evaluated using dog urine. In this study, we answered the questions whether this LFIA can be transferred to human urine samples, and whether this LFIA can be used as a supporting tool prior to LC-HRMS/MS confirmation. Result interpretation by eye and using digitally acquired pixel intensity ratios was investigated with respect to analytical sensitivity. The LFIA detects amatoxins in human urine after visual evaluation to as little as 5 ng/mL (α-amanitin-10 ng/mL, β-amanitin-50 ng/mL, γ-amanitin-5 ng/mL). After digital analysis, pixel intensity ratios were determined to evaluate the LFIA as positive, negative, or trace result. Detection limits were redefined ranging from 1 ng/mL (α- and γ-amanitin) to 3 ng/mL (β-amanitin). For the proof-of-concept, 73 human urine samples submitted to the authors´ laboratory for toxicological analysis were analyzed using the LFIA and LC-HRMS/MS. Only three out of 73 urine samples were tested false positive with the LFIA as LC-HRMS/MS confirmation revealed no detection of amatoxins. Sixteen urine samples were evaluated as trace results and confirmed negative using LC-HRMS/MS except for one case which was positive for α-amanitin but negative for β-amanitin. Although particularly positive and trace results of the LFIA still need to be confirmed, the negative LFIA results correlated well with LC-HRMS/MS.
From promise to practice: why HRMS has yet to fully revolutionize forensic toxicology.
Rodda LN, Ellefsen KN, Mardal M … +5 more , Stockham P, Steuer AE, Mata D, Krotulski AJ, Sarkisian M
J Anal Toxicol · 2025 Sep · PMID 40314114 · Publisher ↗
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