Drug Test Anal [JOURNAL]
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200 papers
Infant Death due to Cannabis Ingestion.
Favretto D, Cirnelli A, Pertile R … +7 more , Stimamiglio R, Cestonaro C, Cuman O, Pagliaro A, Mattiazzi F, Basso C, Galeazzi M
Drug Test Anal · 2025 Oct · PMID 40399755 · Full text
A child died in the emergency room of a local hospital a few hours after ingesting a substance the color of cork and the consistency of earth. At home, a modest amount of resinous substance was found. At the hospital, th...
A child died in the emergency room of a local hospital a few hours after ingesting a substance the color of cork and the consistency of earth. At home, a modest amount of resinous substance was found. At the hospital, the child exhibited alterations in walking, balance, and consciousness. Intubation was needed for the onset of dyspnea, so fentanyl and ketamine were administered during the procedure. A sample of blood was also collected for clinical investigation. During the autopsy, cadaveric samples were collected. Autopsy evaluation revealed multiorgan congestion in the brain, lung, liver, and kidney. Histological investigations were inconclusive. A thorough toxicological investigation was undertaken by immunochemical technique, gas chromatography-mass spectrometry (GC-MS), and liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) on samples of blood, bile, urine, organs, gastric content, and head hair. Toxicological analysis detected cannabinoids, ketamine, norketamine, and fentanyl in blood drawn from the emergency room. Cannabinoids were also observed in postmortem central blood, peripheral blood, urine, bile, brain, lung, and liver samples. Hair analysis showed tetrahydrocannabinol, cannabidiol, cannabinol, methadone and metabolites, cocaine and metabolites, ketamine, morphine, 6-monoacetylmorphine, and fentanyl. Gastric content revealed traces of cannabis products. Acute cannabis intoxication in the context of chronic exposure to numerous drugs has been considered responsible for the death. An increasing number of intoxication cases are being reported worldwide due to the legalization of cannabis. In most cases, these are adults suffering from preexisting conditions, whereas data on younger individuals are still scarce. In this paper, the case of a child who died from acute intoxication due to ingestion of hashish is presented.
Doping Control of Ranitidine in Horses.
Ho HSM, Mizzi JX, Ho ENM … +1 more , Wong WT
Drug Test Anal · 2025 Oct · PMID 40394757 · Publisher ↗
Ranitidine is a histamine H-receptor antagonist commonly used to treat gastric ulceration in horses. The author's laboratory conducted a study some years ago in the early 2000s on its metabolism as well as its urinary el...
Ranitidine is a histamine H-receptor antagonist commonly used to treat gastric ulceration in horses. The author's laboratory conducted a study some years ago in the early 2000s on its metabolism as well as its urinary elimination profile in two geldings. With the technology advancement as well as popularity of blood for doping control testing, the laboratory has recently conducted another administration trials of the substance in six horses to study the in vivo metabolism of ranitidine, aiming to identify and reinvestigate the appropriate target(s) for controlling misuse of ranitidine in horses as well as to study its elimination in blood. To study the elimination and biotransformation of ranitidine, administration experiments were performed by giving six castrated horses (geldings) each 25 mL of Ulcerguard oral paste (equivalent to 9.8 g of ranitidine) in the morning and 20 mL of oral paste (equivalent to 7.9 g of ranitidine) in the afternoon daily for eight consecutive days. The postulated in vivo metabolites included ranitidine-S-oxide (M1), ranitidine-N-oxide (M2), desmethylranitidine (M3a/b) and furoic acid analogue of ranitidine (M4), resulting from oxidation, demethylation and oxidative deamination of ranitidine. To control the misuse of ranitidine in horses, elimination profiles of urinary and plasma ranitidine were established. Free ranitidine was detectable for at most 8 days and 72 h in urine and plasma, respectively. Both metabolites ranitidine-S-oxide and ranitidine-N-oxide were detected for 8 days, and therefore, they could be monitored alongside the parent drug as evidence that the substance has gone through the horse's body.
Metabolic Identification Based on Proposed Mass Fragmentation Pathways of the Anabolic Steroid Bolasterone by Gas Chromatography Tandem Mass Spectrometry.
Muresan AR, Rahaman KA, Rafique FB … +3 more , Son J, Kang MJ, Kwon OS
Drug Test Anal · 2025 Oct · PMID 40386947 · Full text
Bolasterone (7α,17α-dimethyl-androsta-4-en-17β-ol-3-one) was registered on the World Anti-Doping Agency's Prohibited list of substances. This study was aimed at evaluating the metabolism of bolasterone through in vitro (...
Bolasterone (7α,17α-dimethyl-androsta-4-en-17β-ol-3-one) was registered on the World Anti-Doping Agency's Prohibited list of substances. This study was aimed at evaluating the metabolism of bolasterone through in vitro (liver microsomes) and in vivo (rat urine) experiments to propose mass fragmentation pathways of the metabolites by gas chromatography-quadrupole tandem mass spectrometry (GC-EI-MS/MS). Their plausible chemical structures were suggested based on their fragmentation pathways to overcome the lack of available authentic standards. A total of 12 metabolites (5 mono-hydroxylated M1 to M5 and 7 di-hydroxylated M6 to M12) after trimethylsilylation were observed. Key diagnostic ions included m/z 403 (mono-hydroxylated) and m/z 491 (di-hydroxylated) with m/z 143, indicating an intact D ring (M1 to M5, M7, M9, M10, M11). Hydroxylation at the D ring (M6, M12) was characterized by ions m/z 231 or 219. Hydroxylation at the A (M5, M7) or B (M2/M3, M10) rings corresponded to m/z 281 and hydroxylation at C12 of the C ring (M4, M10) was indicated by m/z 285. Based on the comparison with bolasterone analogues such as testosterone and methyltestosterone and the interpretation of fragmentation pathways, the mono-hydroxylation metabolites M1 (at C11), M2/M3 (at C6), M4 (at C12), M5 (at C2), and di-hydroxylation metabolites M6 (at C11 and C16), M7 (at C2 and C11), M10 (at C6 and C12), and M12 (at C12 and C16) were proposed. The hydroxylation sites of M8, M9, and M11 could not be determined. This data can be useful for identifying hydroxylated metabolites by interpreting mass spectra of anabolic steroids with no standards available.
Identification of the Novel Synthetic Opioid N-Pyrrolidino Isotonitazene at an Australian Drug Checking Service.
Curtis B, Lawes DJ, Caldicott D … +1 more , McLeod MD
Drug Test Anal · 2025 Oct · PMID 40384477 · Full text
2-Benzylbenzimidazole opioids and related derivatives, also known as 'nitazenes', present a growing threat to public health. Emerging in Europe in 2019, the nitazene group of drugs is a recent addition to the novel synth...
2-Benzylbenzimidazole opioids and related derivatives, also known as 'nitazenes', present a growing threat to public health. Emerging in Europe in 2019, the nitazene group of drugs is a recent addition to the novel synthetic opioid class and has been associated internationally with adverse effects in drug users, overdose clusters and significant mortality. The high potency of many nitazene derivatives, which can in many cases exceed that of fentanyl, poses a significant challenge to the public health and early warning systems used to detect and respond to the emergence of new high-risk substances. This report describes close collaboration between an Australian drug checking service and a nearby university laboratory to identify and characterise the novel synthetic opioid N-pyrrolidino isotonitazene in an expected oxycodone sample presented by a member of the public. Though no prior publications are available describing the presence of this nitazene in the drug market, previously reported in vitro evaluation of this compound reveals it to be among the most potent nitazene opioid agonists known. The study highlights the rapid response possible though engaging drug users with drug checking services as a market monitor and early warning system to alert health services and the broader community to the presence of unexpected, high-risk substances. Integration of well-resourced and supported drug checking services provides a powerful approach to tackle the public health threats associated with novel synthetic opioids and other drugs of concern.
Chemical Analysis and Alkaloid Intake for Kratom Products Available in the United States.
Sharma A, Smith KE, Kuntz MA … +8 more , Berthold EC, Elashkar OI, Guadagnoli N, Kanumuri SRR, Mukhopadhyay S, Panlilio LV, Epstein DH, McCurdy CR
Drug Test Anal · 2025 Oct · PMID 40377101 · Publisher ↗
Previously, we used ecological momentary assessment (EMA) to evaluate motivations and temporal patterns of kratom use for 15 days among US adult kratom consumers (N = 357). Here we present the content analyses of the pro...
Previously, we used ecological momentary assessment (EMA) to evaluate motivations and temporal patterns of kratom use for 15 days among US adult kratom consumers (N = 357). Here we present the content analyses of the products used during that nationwide study, with quantification of 10 kratom alkaloids. The samples (N = 341) were primarily whole-leaf products, not extracts, and were similar to each other in their alkaloid composition, closely matching the chromatographic-mass spectrometry fingerprint expected for Mitragyna speciosa leaf material. We found no evidence of adulteration with illicit or prescription drugs. With participants' self-reported data on kratom amount per use, we calculated mitragynine intake per use: mean 31.3 mg and median 25.4 mg (range 2.0-205.9 mg). With self-reported data on frequency, we calculated mitragynine intake per day, it ranged from 78.3 to 134.6 mg (mean) or 50.8 to 101.6 mg (median). This is the most comprehensive analysis of US whole-leaf kratom products to date. The coupling of self-report and product sample-analysis data to quantify daily alkaloid intake is foundational for designing controlled clinical trials of kratom in healthy volunteers. These findings on kratom products' chemical composition and daily kratom alkaloid consumption can also inform clinicians, policymakers, and consumers, particularly for whole-leaf material.
Growing Trend of Novel or Experimental Substances Not Approved for Human Use Sold as Consumer Products Poses Threat to Athletes, Service Members, and Public Health.
Eichner A, Lindsey AT, Coyles J
Drug Test Anal · 2025 Oct · PMID 40377069 · Publisher ↗
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Exposure to Drugs of Abuse in Children and Adolescents Investigated by Hair Analysis.
Cestonaro C, Terranova C, Carollo M … +3 more , Russo A, Aprile A, Favretto D
Drug Test Anal · 2025 Oct · PMID 40374543 · Publisher ↗
Hair testing is a very effective method for drug use investigation. The finding of drugs of abuse in children hair may reflect environmental exposure, accidental ingestion, passive inhalation, intentional administration,...
Hair testing is a very effective method for drug use investigation. The finding of drugs of abuse in children hair may reflect environmental exposure, accidental ingestion, passive inhalation, intentional administration, and, in case of neonates and infants, in-utero and breast exposure. Despite its increasing use, interpretation of children hair analysis remains difficult, cut-off values used in adults cannot be applied, and no reference ranges for different age groups currently exist. To contribute to the identification of a reference framework for hair analysis in children and adolescents, a comparison of data gathered from different geographical areas could be useful. With this view, this study shows the results of hair analysis of children and adolescents who underwent testing for clinical purposes in a hospital setting in north-east Italy. Cocaine represents the most frequently detected drug in all age groups (overall, 94 positives). The highest number and ratio of cocaine positivity was found among children aged 1-3 years (21 out of 32 children; 65.6%) and the highest concentration among infants within 1 year. The results suggest that exposure to drugs of abuse represents a nonnegligible problem particularly in infants and toddlers, which requires special attention in clinical and social setting. In view of the multiple possible ways of exposure, it is essential to perform a case-by-case evaluation and to collect as much information as possible to frame the case and to implement the most effective child protection measures.
Surface Wipe Sampling of Hazardous Medicinal Products: A European Interlaboratory Comparison Study.
van den Berg RB, Korczowska E, Santos MSF … +9 more , Portilha-Cunha MF, Ribeiro ARL, Bláhová L, Bláha L, Eyser CV, Tuerk J, van Rossen RCJM, Wilms EB, Crul M
Drug Test Anal · 2025 Oct · PMID 40324795 · Full text
Workplace monitoring of hazardous medicinal products (HMPs) using surface wipe sampling is becoming common practice in many European hospitals and pharmacies. However, no independent quality control is available to valid...
Workplace monitoring of hazardous medicinal products (HMPs) using surface wipe sampling is becoming common practice in many European hospitals and pharmacies. However, no independent quality control is available to validate wiping procedures and analytical methods. This study aimed to conduct a Europe-wide interlaboratory comparison (ILC) program to independently and blindly assess laboratory performance and variability in HMP detection. Four European laboratories participated in the study. Six HMPs-cyclophosphamide, etoposide, gemcitabine, ifosfamide, methotrexate, and paclitaxel-were prepared at four concentrations (5000, 2000, 200, and 20 ng/mL) and applied to a 400-cm stainless-steel surface, then wiped by the coordinating body according to each laboratory's protocol. Wipe samples were distributed to individual laboratories, where blind analyses were conducted. Target criteria for accuracy and recovery were set at 70%-130% and 50%-130%, respectively. Of the 80 samples, 69 (86%) met accuracy targets, and 70 (88%) met recovery targets. Accuracy was often overestimated for the lowest concentrations of cyclophosphamide, etoposide, methotrexate, and paclitaxel by Laboratory A. Laboratory D showed low accuracy for paclitaxel at three lower concentrations. Among the 10 samples that did not meet recovery targets, all were below 50% and involved etoposide and paclitaxel. This ILC program demonstrates a viable method for evaluating laboratory performance in HMP detection, offering an external validation mechanism for surface wipe sampling methods. A future goal is to establish a global ILC program with a designated coordinating body for managing it effectively.
Gel Electrophoretic Detection of Black Market ACE-031.
Reichel C, Filip T, Gmeiner G … +1 more , Thevis M
Drug Test Anal · 2025 Oct · PMID 40312924 · Full text
The usage of ACE-031 (Ramatercept), a dimeric fusion protein consisting of a human activin receptor IIB (ACVR2B) fragment linked to an Fc-part of human IgG1, is banned according to chapter S4.3 of the "WADA 2024 List of...
The usage of ACE-031 (Ramatercept), a dimeric fusion protein consisting of a human activin receptor IIB (ACVR2B) fragment linked to an Fc-part of human IgG1, is banned according to chapter S4.3 of the "WADA 2024 List of Prohibited Substances and Methods" due to its potential performance enhancing properties. While ACE-031 has not yet been pharmaceutically approved, it is sold as research chemical on the "black market" (BM). The article presents a study on BM ACE-031 products and its detection by gel-electrophoresis and Western blotting. Of 14 tested products, only 12 contained an ACVR2B-immunoreactive protein. Electrophoretic separation by SDS-PAGE also showed that the 12 ACVR2B-products contained many other proteins in addition to the main compound (ca. 58.4 kDa). Further analyses by mass spectrometry and immunoblotting revealed that the 12 products contained the full-length human activin receptor IIB instead of ACE-031. The absence of an Fc-fusion protein was further confirmed by treatment with IdeS protease, which was unable to cleave the BM products. In addition, it was demonstrated that the protocol we developed to detect luspatercept (another ACVR2B-Fc fusion protein) in human serum could also be successfully applied for the detection of BM ACE-031. Because administering black market products to human subjects was not ethically justifiable, a study was conducted with rats. In rat serum, BM ACE-031 was detectable up to 48 h post administration. However, due to the relatively high dose applied (10 mg/kg body weight) and possible differences in metabolism, the detection window may be different in humans.
Surveillance of Anabolic Agent Residues in US Meat Supply by Liquid Chromatography With High-Resolution Tandem Mass Spectrometry.
Snethen CM, Fedoruk MN, Ahrens ED … +3 more , Sobolevskii TG, Avliyakulov NK, Johnson BJ
Drug Test Anal · 2025 Oct · PMID 40312815 · Publisher ↗
This study provides data on the prevalence of animal growth promoter residues through a comprehensive US surveillance approach targeting multiple species via retail purchases. Over a year, residue levels were analyzed in...
This study provides data on the prevalence of animal growth promoter residues through a comprehensive US surveillance approach targeting multiple species via retail purchases. Over a year, residue levels were analyzed in beef, pork, and poultry samples from eight US cities, screening for a panel of anabolic steroids, other anabolic agents, selective androgen receptor modulators (SARMs), and non-prohibited controls using sensitive high-resolution tandem mass spectrometry. Thirteen of the 53 beef samples tested positive for ractopamine, with a peak concentration of 14 ng/g in liver-well below CODEX and FDA maximum residue limits (MRLs). Trenbolone-17β and estradiol were also found in beef but remained within MRLs, indicating no health risk. In addition, pork samples showed minimal contamination, with just one pork kidney testing positive for nandrolone (0.08 ng/g) under the limit of detection and estradiol (0.5 ng/g), likely from endogenous hormone production in intact male pigs. Pork muscle and liver samples were residue-free. Chicken samples showed limited residues, with estradiol detected in three out of 35 muscle samples at 0.01 ng/g. These findings highlight the effectiveness of regulatory practices in limiting growth promoter residues in commercial meat, affirming that residue levels in meat products remain within regulatory limits and reinforces consumer safety and confidence.
Pooled Sampling Technique to Improve the Monitoring of Medication Use in the Racehorse Industry.
Chambers A
Drug Test Anal · 2025 Oct · PMID 40296484 · Publisher ↗
All anti-doping programmes face financial constraints and monitoring trends in medication use or abuse in a population of racehorses can be difficult and expensive. Obtaining biological samples is the primary method of a...
All anti-doping programmes face financial constraints and monitoring trends in medication use or abuse in a population of racehorses can be difficult and expensive. Obtaining biological samples is the primary method of anti-doping control in individual horses or stables of horses but can be invasive and expensive. Another important practice of anti-doping control has been the confiscation of used and filled syringes by regulators for individual forensic analysis. Pooled samples testing involves the testing of multiple individual samples together as one composite sample. This pooled sample approach has been employed to gather information concerning populations' exposure to substances and infectious agents including the analysis of samples of wastewater (a large, pooled sample) that have been used during the pandemic to monitor the presence of new COVID variants. Moreover, pooled samples of urine and wastewater have been used to monitor for recreational drug use and for the presence of new psychoactive substances in cities and large events. This approach has been credited with providing timely insight in the trends of illicit drugs use. To be effective, an anti-doping programme should not be predictable to avoid being defeated by countermeasures; therefore, the implementation of new methods is considered essential. A new pooled sampling technique using confiscated groups of used syringes and needles including biomedical sharps containers obtained from veterinarians and other horse racing industry participants has been employed over several years in Ontario, Canada. These containers held needles used to administer substances to racehorses along with syringes and other debris. The analysis of the wash provided a timely insight of medications being administered in horses, and substances present at racetracks and training centres including substances predominantly of human use and abuse. Sharp containers confiscated from veterinarians and trainers provided insight into injectable medications administered at numerous stables and to hundreds of horses.
Transcriptomic Biomarkers in Blood Indicative of the Administration of Recombinant Human Erythropoietin to Thoroughbred Horses.
Cheung HW, Wong KS, Cheng PCF … +4 more , Tsang CYN, Farrington AF, Wan TSM, Ho ENM
Drug Test Anal · 2025 Oct · PMID 40256823 · Publisher ↗
Erythropoiesis-stimulating agents (ESAs) continue to be a significant threat to the integrity of human and equine sports. Besides conventional direct testing, monitoring the biomarkers associated with the effects of ESAs...
Erythropoiesis-stimulating agents (ESAs) continue to be a significant threat to the integrity of human and equine sports. Besides conventional direct testing, monitoring the biomarkers associated with the effects of ESAs may provide a complementary approach via indirect detection to enhance doping control. In this study, we applied RNA-sequencing (RNA-seq) to discover blood RNA biomarkers in Thoroughbred horses after administration with a long-acting form of recombinant human erythropoietin (rhEPO), methoxy polyethylene glycol epoetin beta, Mircera®. A single subcutaneous administration of Mircera® at ~ 4.2 μg/kg was effective in elevating haematocrit, haemoglobin and erythrocyte levels to varying extents in as early as 4 days post-administration in all three horses, which persisted for 40 days post-administration (the last sample collected). RNA-seq was applied to analyse blood transcriptomic changes. Differential gene expression analysis has allowed the identification of 46 genes that showed dramatic and temporary upregulation at 4-11 days after Mircera® administration. STRING analysis has identified the functional enrichment of 15 genes important for erythropoiesis and erythrocyte function, supporting the idea of an increased release into the peripheral circulation of residual RNA-containing reticulocytes after rhEPO exposure, which would otherwise mature normally inside the bone marrow in horses. Machine learning of blood transcriptomes has enabled the discrimination of samples with or without Mircera administration. Therefore, our study has provided new insights into the biological mechanism of erythropoiesis caused by rhEPO administration in horses and has provided evidence supporting the control of misuse of ESAs by monitoring the equine blood transcriptome.
Endogenous Nature of Hydrocortisone Acetate in Horse.
Kong FKW, Wong ASY, Wan TSM … +1 more , Ho ENM
Drug Test Anal · 2025 Oct · PMID 40230061 · Publisher ↗
Hydrocortisone acetate, a synthetic acetate ester of hydrocortisone, was detected in horse blood samples collected from Thoroughbreds. Hydrocortisone acetate is generally considered an indicator for exogenous administrat...
Hydrocortisone acetate, a synthetic acetate ester of hydrocortisone, was detected in horse blood samples collected from Thoroughbreds. Hydrocortisone acetate is generally considered an indicator for exogenous administration in horses. As hydrocortisone acetate has been previously reported to be endogenous in selected mammals, a proof-of-concept study was performed to evaluate the possible endogenous nature of hydrocortisone acetate in horses by in vitro incubation experiments using homogenized horse brain tissue.
A Review of Hypoxen Pharmacology and Potential to Enhance Sports Performance.
Jędrejko K, Catlin O, Faiss R … +1 more , Pokrywka A
Drug Test Anal · 2025 Oct · PMID 40223246 · Publisher ↗
Pharmacological potential of Hypoxen, previously registered as Olifen is evaluated herein. Hypoxen is categorized as antihypoxic agent. The active substance is polydihydroxyphenylene thiosulfonate sodium. Human studies a...
Pharmacological potential of Hypoxen, previously registered as Olifen is evaluated herein. Hypoxen is categorized as antihypoxic agent. The active substance is polydihydroxyphenylene thiosulfonate sodium. Human studies are limited and no clinical trials following international standards is available. There is however a developed body of knowledge emerging from original studies conducted by the Russian Military Medical Academy in 1980s and 1990s despite limited online access. Hypoxen is promoted to improve oxygen supply or reduce oxygen consumption under hypoxic conditions and physical load. It is thought to support faster recovery, and can be used in complex treatments of diseases accompanied by hypoxia like myocardial ischemia. From clinical perspective, it may enhance cellular respiration by improving coupling in the respiratory chain/accelerating oxidative phosphorylation, but also inhibit succinate dehydrogenase (SDH), and activate mitochondrial ATP-sensitive potassium channels (mitoK) in skeletal muscles and myocardium. In 2023, the World Anti-Doping Agency (WADA) added Hypoxen to the Monitoring Program as there had been documented evidence of its use by athletes. On in vitro experiments compared the influence of Hypoxen on oxidative phosphorylation with mitochondrial uncoupling agent 2,4-dinitrophenol (DNP) a unique metabolic modulator that strongly accelerates the metabolism rate, prohibited since 2024 by WADA. Most studies focus on exercise performance, and may provide some evidence that Hypoxen has the potential to enhance performance, the first criteria considered for addition of substance to the WADA Prohibited List. Pharmacodynamics and ergogenic effects of Hypoxen suggests potential as metabolic modulator.
Optimization of Parallel Artificial Liquid Membrane Extraction for the Determination of Over 50 Psychoactive Substances in Oral Fluid Through UHPLC-MS/MS.
Croce M, Montesano C, Bracaglia I … +4 more , Bartolini F, Mascini M, Compagnone D, Sergi M
Drug Test Anal · 2025 Oct · PMID 40211089 · Full text
Over the past decade, there has been a diversification of the psychoactive substances available among drug users, resulting in the expansion of a dynamic market of synthetic molecules that are challenging for drug of abu...
Over the past decade, there has been a diversification of the psychoactive substances available among drug users, resulting in the expansion of a dynamic market of synthetic molecules that are challenging for drug of abuse testing. Multiclass analytical methods are useful to deal with these new psychoactive substances (NPS), but sample preparation can be difficult and generate significant amounts of chemical waste. The aim of this work was the development of a high-throughput microextraction method for the determination of 56 drugs belonging to different pharmacological classes in oral fluid (OF), including both traditional drugs and NPS. In the proposed workflow, the OF sample is cleaned-up by parallel artificial liquid membrane extraction (PALME) and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Two hundred microliters of OF are mixed with 1800 μL of carbonate buffer 0.5 M (pH 12) and 0.4 g of sodium chloride and inserted into a donor plate; the acceptor plate embed a dodecylacetate-supported liquid membrane and an acceptor solution composed of 50 μL formic acid 0.1% in H2O: MeOH, 80:20 (v/v); the whole assemblage is placed on an orbital shaker for 120 min for extraction. A full factorial design has been employed for extraction optimization to make it suitable for LC-MS/MS. The developed method is an example of green chemistry and may be used for screening and quantitative purposes, with limits of detection ranging from 0.01 to 1.5 ng mL and optimal performance in term of precision and accuracy for 49 out of 56 drugs tested.
Evaluating the Early Operational Performance of the Endocrine Module in the Athlete Biological Passport.
de Figueiredo M, Marchand A, Rhumorbarbe D … +5 more , Mann R, Saugy J, Robinson N, Lobigs LM, Ericsson M
Drug Test Anal · 2025 Oct · PMID 40195547 · Publisher ↗
In collaboration with various Anti-Doping Organisations and the Paris WADA-accredited laboratory, the International Testing Agency spearheaded the implementation of the Endocrine Module ahead of the Olympic Games Paris 2...
In collaboration with various Anti-Doping Organisations and the Paris WADA-accredited laboratory, the International Testing Agency spearheaded the implementation of the Endocrine Module ahead of the Olympic Games Paris 2024. This article presents a data-driven evaluation of its early integration within Athlete Biological Passport (ABP) programs. The assessment of endocrine profile testing follows the complete ABP sample lifecycle, from collection to the submission of reports by the Athlete Passport Management Unit. The article identifies both challenges and opportunities by integrating testing, analytical and ABP perspectives and offers recommendations for the future development of the Endocrine Module. Furthermore, it explores the Endocrine Module's synergy with the Haematological and Steroidal Modules emphasising on key areas for the enhancement of ABP programs as a whole.
Sports Supplement Analysis Survey for the Prevalence of WADA Prohibited Substances in the Australian Online Marketplace.
Barker L, Cawley A, Speers N … +2 more , Knowler K, Chilman K
Drug Test Anal · 2025 Oct · PMID 40180440 · Full text
Sports supplements are used extensively by people involved in competitive sports. However, a proportion of sports supplements contain World Anti-Doping Agency (WADA) Prohibited Substances, which have the potential to cau...
Sports supplements are used extensively by people involved in competitive sports. However, a proportion of sports supplements contain World Anti-Doping Agency (WADA) Prohibited Substances, which have the potential to cause an anti-doping rule violation (ADRV). In 2022, Sport Integrity Australia (SIA) contracted Human and Supplements Testing Australia (HASTA) to purchase and analyse 200 sports supplement products available in the Australian online marketplace. The aim of the survey was to determine the likelihood of purchasing a product containing one or more WADA Prohibited Substances. Of the 200 products purchased, 35% contained WADA Prohibited Substances, with the majority of these findings being stimulants. Products marketed as pre-workouts, fat burners and muscle builders were the most likely to contain prohibited substances. It is notable that 83% of the WADA Prohibited Substances detected were naturally occurring compounds; only two of the products contained high levels of synthetic stimulants. Fifty-seven percent of the products that contained WADA Prohibited Substances did not have the substances labelled as ingredients on the packaging or website. Athlete education concerning sports supplements should remain a high priority, with a prevalence rate of approximately one in three products purchased containing WADA Prohibited Substances. To reduce the risk of an inadvertent ADRV, if an athlete wishes to use sports supplements, they should consult with their medical practitioner or accredited sports dietician (ASD) as to whether they require them and, if so, only take products that have been independently tested.
Romancing the Three States of Methamphetamine.
Chan SJ, Tan-Lee NL, Lui CP … +1 more , Moy HY
Drug Test Anal · 2025 Oct · PMID 40174902 · Publisher ↗
This study evaluates the methamphetamine contamination routes through exposure to sweat, smoke in the environment and direct contact, as well as the effectiveness of decontamination procedures. For sweat exposure, drug-f...
This study evaluates the methamphetamine contamination routes through exposure to sweat, smoke in the environment and direct contact, as well as the effectiveness of decontamination procedures. For sweat exposure, drug-free hair samples (n = 10) were sprayed with artificial sweat containing amphetamine and methamphetamine at either 'low' (2.6 and 6.5 ng/mL, respectively) or 'high' concentration (3.1 and 25.2 ng/mL, respectively), 5 times (2 mL each time) per day for 5 days. In the smoke exposure, 2 mg of methamphetamine hydrochloride powder was heated and vapourised on one side of an enclosed cardboard box containing hair (n = 10) on the other side. In the direct contact with methamphetamine powder, 0.6 mg of methamphetamine hydrochloride was rubbed gently by hand onto hair (n = 5) for 1 min. Each contaminated hair sample was then subjected to either one of the 3 wash methods, namely, (1) sequential wash using dichloromethane, water, and methanol; (2) sequential wash by hexane and acetone; and (3) methanol wash only. All solvents and hair samples were analysed. The results show that it was relatively difficult to remove methamphetamine from the hair that was exposed to methamphetamine in artificial sweat, followed by methamphetamine smoke and then direct contact with the methamphetamine powder. None of the wash methods was able to completely remove amphetamine and methamphetamine from the contaminated hair. Overall, methods (1) and (3) show better decontamination efficacy than method (2), probably due to the polar protic nature of the solvents.
Investigations Into Structures of In Vitro-Derived Phase I Metabolites of a Novel 20-Keto-Steroid S42 by GC-EI HR MS Analysis and Chemical Synthesis.
Wen HC, Wagener F, Piper T … +3 more , Neudörfl J, Thevis M, Schäfer M
Drug Test Anal · 2025 Sep · PMID 40159391 · Publisher ↗
S42, 4-Methyl-19-norpregna-1,3,5(10)-trien-20-one a new 20-keto-steroid, is a novel selective androgen receptor modulator (SARM). The World Anti-Doping Agency (WADA) bans the use of SARMs in sports at all times. In prepa...
S42, 4-Methyl-19-norpregna-1,3,5(10)-trien-20-one a new 20-keto-steroid, is a novel selective androgen receptor modulator (SARM). The World Anti-Doping Agency (WADA) bans the use of SARMs in sports at all times. In preparation of a sensitive detection procedure to control for S42 abuse, in vitro metabolism experiments were conducted and biotransformation products were analyzed with GC-EI MS-orbitrap instrumentation. S42-C20-OH, S42-C6ß-OH, and S42-C7α-OH were synthesized as reference material to study their exemplary EI-HR (electron ionization- high resolution) mass spectra. Additionally, S42-d7, synthesized earlier with H-labels at carbon atoms C1, C2, C3, C6, and C7, was used for the in vitro metabolism study. Comparison of the respective mass spectra of labeled and unlabeled reference materials and of specifically mass-shifted fragment ions provided the foundation for the structure elucidation of S42 in vitro phase I metabolites. Molecular ions of selected S42 phase I metabolites found in the in vitro experiments were submitted to higher energy collisional dissociation (HCD) MS-product ion experiments to allow straightforward and secured assignment and interpretation of fragmentation patterns. At least eight phase I metabolites of S42 were identified in the in vitro study and analyzed as tri-methyl-silyl ether derivatives. Specifically, different singly, doubly, and triply hydroxylated metabolites of S42 were identified and analyzed with GC-EI HR MS.
Analysis of Testosterone Esters in Serum and DBS Samples-Results From an Interlaboratory Study.
Langer T, Musenga A, Stojanovic B … +9 more , Pecher D, Gmeiner G, Harju L, Suominen T, Mongongu C, Ericsson M, Grabherr S, Kuuranne T, Nicoli R
Drug Test Anal · 2025 Sep · PMID 40138730 · Publisher ↗
Testosterone (T) formulations that are used for doping purposes often contain the steroid in esterified forms. As these esters are hydrolysed in the bloodstream before renal excretion, they can be detected in blood matri...
Testosterone (T) formulations that are used for doping purposes often contain the steroid in esterified forms. As these esters are hydrolysed in the bloodstream before renal excretion, they can be detected in blood matrices and have not been detected in urine so far. Serum samples can additionally be used for longitudinal blood steroid profiling, but their collection, shipping and storage have some disadvantages. The use of dried blood spots (DBS), an alternative blood matrix, is more convenient for pre-analytical and post-analytical aspects but is not fully established in antidoping laboratories yet. To evaluate the ability of multiple antidoping laboratories to detect T-esters in serum and DBS samples, an interlaboratory study was organised. Common T-esters were spiked in five samples of each matrix (serum, cellulose card DBS, polymeric DBS) at concentrations that correspond to an administration scenario and sent as blinded specimens to each laboratory. The laboratories were requested to apply their own analytical method to detect the T-esters and to provide a rough estimate of their concentrations. All laboratories identified the spiked testosterone esters correctly in all samples and the estimated concentrations were deemed comparable (average relative standard deviation < 30%), considering that only qualitative initial testing procedures (ITPs) were used. This study could firstly demonstrate the capability of different analytical approaches to analyse T-esters in serum and DBS samples and, secondly, show that the methods employed by the participating laboratories are all fit for purpose.
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