Drug Test Anal [JOURNAL]
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200 papers
Statistical Discrimination of Urinary Steroid Biomarkers in the Athlete Biological Passport: A Novel Approach to an Abnormal Steroid Profile Score (ASPS).
Hopker JG, Griffin JE, Fedoruk MN … +1 more , Lewis LA
Drug Test Anal · 2026 May · PMID 41785482 · Full text
The steroidal module of the Athlete Biological Passport (ABP) longitudinally monitors five ratios between urinary concentrations of endogenous anabolic and androgenic steroids. Even though it has improved detection of te...
The steroidal module of the Athlete Biological Passport (ABP) longitudinally monitors five ratios between urinary concentrations of endogenous anabolic and androgenic steroids. Even though it has improved detection of testosterone doping, the interpretation of data from multiple discrete biomarkers is complex. This study sought to create a single score to identify doping rather than relying on the interpretation of each parameter alone. A Bayesian model was used to define an ABP sequence probability for each biomarker to assess the extremity of a measurement relative to the expected levels from ABP. This was used to discriminate between doped and presumed clean individuals based upon pattern classification of biomarkers using classification algorithms. Data were obtained from laboratory-controlled experimental studies as well as routine doping control tests. A laboratory model (where classifier is trained using the laboratory-controlled data only) and a mixed model (where classifier is trained on combined laboratory-controlled and doping control data) were developed and tested on the doping control data. Logistical regression was seen to have the best classification performance across the methods used, with the Abnormal Steroid Profile Score (ASPS) representing the estimated probability from the logistical regression model. Classifier performance produced an AUC of 0.67 and 0.75 when trained on the laboratory model and the mixed model, respectively, with T/E and 5α-Diol/5β-Diol representing the main biomarkers driving the ASPS. These findings demonstrate that the ASPS can discriminate between the doping status of individuals, even if a mixture of steroids, administration methods and doses are used.
Concentration and Detection Time of Nitrous Oxide in Blood Following Controlled Inhalation.
Lindholm AØ, Nielsen MKK, Kristensen M … +3 more , Larsen MH, Heiberg J, Rasmussen BS
Drug Test Anal · 2026 Apr · PMID 41765373 · Full text
Recreational use of nitrous oxide (laughing gas, NO) is becoming increasingly common and abuse is often seen in relation to driving, posing significant traffic-safety concerns. Only a few studies exist on blood concentra...
Recreational use of nitrous oxide (laughing gas, NO) is becoming increasingly common and abuse is often seen in relation to driving, posing significant traffic-safety concerns. Only a few studies exist on blood concentrations of NO and its detectability over time after use. In this study, 11 volunteers received controlled administration of 50% NO for 10 min, after which blood samples were drawn and analyzed for NO by headspace-gas chromatography-mass spectrometry (HS-GC-MS). Pharmacokinetic modelling indicated that elimination of NO can best be described by a two-compartment model with half-lives of 2.4 and 31 min. Although both the pharmacological effect and intoxication typically disappear within minutes after intake, NO remained detectable in blood for an average of 62 min at a cutoff of 0.2 mL/L and 132 min at a cutoff of 0.05 mL/L.
Urine or Oral Fluid for Random Testing? A Review of Real World Data.
Evers R, Cottrill Z, Kindred P … +4 more , Csete M, Geyser-Wilcox J, Pennington L, Llewelyn P
Drug Test Anal · 2026 Apr · PMID 41742805 · Publisher ↗
Most comprehensive workplace drug testing programmes include random or unannounced testing as a means to deter staff from drug use and mitigate risk. Testing can be performed on urine or oral fluid (saliva) either by on-...
Most comprehensive workplace drug testing programmes include random or unannounced testing as a means to deter staff from drug use and mitigate risk. Testing can be performed on urine or oral fluid (saliva) either by on-site lateral flow tests that give an immediate negative, or a not-negative/unconfirmed result, or by return of the full sample to the laboratory for screening. Any sample, whether screened on site or at the laboratory, is then confirmed using a sensitive and specific method that can prove the identity of the drugs present and also determine the difference between related drugs, such as morphine and codeine. This study reviews data from over 12 months to determine the positive rates for urine and oral fluid (saliva) testing. This allows employers to determine the likely effectiveness of their screening policy and estimate the number of staff expected to produce a not-negative/unconfirmed result. The results show that the primary screening results, whether based on urine or oral fluid, are reliable with a positive predictive value of better than 80% for commonly used drugs. Urine testing identifies 8.9% of samples as requiring further confirmation testing, whereas oral fluid (saliva) identifies 3.4%. Urine testing is better where employers wish to ensure a drug-safe workplace, whereas oral fluid (saliva) testing will probably lead to a lower level of disciplinary action.
Evaluation of Therapeutic Opioids in Skin-Derived Matrices (Sweat and Sebum) of Neonatal and Pediatric Patients and Their Role in Opioid Incorporation Into Hair.
Polke M, Zapf F, Scherer JT … +5 more , Voegel CD, Restin T, Schmid MW, Kraemer T, Binz TM
Drug Test Anal · 2026 Apr · PMID 41742792 · Full text
Hair analysis is a valuable tool in forensic toxicology to assess opioid exposure in both adult and pediatric populations. However, interpretation of analysis results is challenged by the scarcity of reliable reference d...
Hair analysis is a valuable tool in forensic toxicology to assess opioid exposure in both adult and pediatric populations. However, interpretation of analysis results is challenged by the scarcity of reliable reference data on opioids in hair and by the lack of knowledge on involved hair incorporation pathways, e.g., via skin-derived matrices such as sweat and sebum. In order to address these limitations, we conducted a clinical study in cooperation with the University Children's Hospital Zurich, where we obtained and analyzed hair and skin swab samples of 150 pediatric intensive-care patients (median age: 53 days). These patients had a treatment history with opioids, including fentanyl, sufentanil, remifentanil, alfentanil, morphine, methadone, and hydromorphone. Most of the substances, as well as selected metabolites, were repeatedly detected in both sample types, exhibiting comparable concentration trends and metabolite-to-parent drug ratios. For fentanyl and morphine, a positive correlation was observed between the administered opioid doses and the analyte concentrations in both the hair and skin swab samples. In addition, a correlation was found for fentanyl between concentrations in hair and skin swab samples (r = 0.356, p = 0.0007), indicating a contribution of skin-derived matrices to opioid incorporation into the hair of children and neonates. The comprehensive and clinical nature of this study provides unique value for the generated concentration data and associated findings, providing a potential reference source for future interpretation of hair analysis results.
Bromazolam Tablet Quantification and Analysis of Post-Mortem Cases From the National Programme on Substance Use Mortality (NPSUM).
Gardner M, Millea MF, Craft S … +7 more , Andrews R, Scott J, Husbands SM, Pudney CR, Sutcliffe OB, Copeland CS, Sunderland P
Drug Test Anal · 2026 Apr · PMID 41732118 · Full text
Bromazolam is a new psychoactive substance (NPS) benzodiazepine commonly identified by drug checking services and in post-mortem toxicological analyses in the United Kingdom, Europe, and North America. At the time of wri...
Bromazolam is a new psychoactive substance (NPS) benzodiazepine commonly identified by drug checking services and in post-mortem toxicological analyses in the United Kingdom, Europe, and North America. At the time of writing, there are no studies that present quantitative analyses of bromazolam in street tablets. Here we describe the first quantitative analysis of bromazolam tablets, from samples submitted by UK drug checking services and police forces between 2022 and 2025. Using validated GC-EI-MS and H NMR methods, 47 tablet samples were quantified revealing a median bromazolam dose of 0.49 mg (interquartile range = 1.02 mg) per tablet, range of 0.09-5.4 mg. Over half of the tablet submissions (55%) mimicked the appearance of licensed pharmaceuticals alprazolam or diazepam, raising significant concerns around mis-selling of street bromazolam tablets and the risks of unintentional high-dose exposure to an NPS compound. To contextualise these findings, we also report post-mortem data from the UK National Programme on Substance Use Mortality (NPSUM), in which bromazolam was detected in 396 drug-related deaths between April 2021 and July 2024. Bromazolam detections in deaths rose from 28 deaths in 2021 to 160 deaths in 2023. Bromazolam was implicated in causing death in 82.8% of cases, with a median post-mortem blood concentration of 43 ng/mL. Notably, bromazolam was co-detected with an average of seven other substances per case, most commonly other central nervous system (CNS) depressants. These findings underscore the public health risks posed by bromazolam as an NPS benzodiazepine and highlight the urgent need for monitoring, harm reduction and forensic toxicology guidance.
The MAIIA EPO Purification Gel Kit, 7D3-An Alternative Purification Kit for Screening and Confirmation Procedures. Validation for Serum and Plasma Matrices.
Hempel J, Schwenke D, Reimann S … +1 more , Voss SC
Drug Test Anal · 2026 Apr · PMID 41732113 · Publisher ↗
In this study, the performance of the MAIIA EPO purification kit 7D3 was evaluated for human serum and plasma matrices. The kit was validated for selectivity in comparison to the MAIIA 3F6 kit, reliability at minimum req...
In this study, the performance of the MAIIA EPO purification kit 7D3 was evaluated for human serum and plasma matrices. The kit was validated for selectivity in comparison to the MAIIA 3F6 kit, reliability at minimum required performance levels (MRPLs), limit of detection (LOD), carryover and recovery to meet compliance with the World Anti-Doping Agency criteria of the technical document 2024 (TD2024EPO). In addition, the influence of haemolysis on plasma samples was evaluated. In total, 30 serum samples and 30 plasma samples were tested for selectivity; 10 serum and 10 plasma samples, for reliability at MRPL and recovery; and six serum and six plasma samples, for LOD. The influence of haemolysis was evaluated by creating mixtures of 0%, 25%, 50%, 75% and 100% of a hemolytic sample and a blank sample, and the same sample spiked at the MRPL level. This study validates that the 7D3 kit meets all WADA TD2024EPO criteria, providing a reliable alternative for the purification of erythropoietin receptor agonists from human serum and plasma matrices in both screening and confirmation procedures in routine antidoping analysis.
Detection of a Distinct Erythropoietin (EPO) Profile After Isoelectric Focusing in Patients With Familial Erythrocytosis.
Martin L, Idriss S, Maaziz N … +7 more , Huerre S, Mekacher LR, Lahmek K, Girodon F, Gardie B, Ericsson M, Marchand A
Drug Test Anal · 2026 Apr · PMID 41713879 · Full text
Since the implementation of the first erythropoietin (EPO) detection method to fight the use of recombinant EPO (rEPO) for doping, the techniques used and identification criteria have evolved. The first technique involve...
Since the implementation of the first erythropoietin (EPO) detection method to fight the use of recombinant EPO (rEPO) for doping, the techniques used and identification criteria have evolved. The first technique involved iso-electric focusing (IEF-PAGE) to separate EPO isoforms by their charges. It can still be used for initial testing but is now largely replaced by electrophoretic separation based on molecular weight (SDS-PAGE or SAR-PAGE) to conclude decisively on the presence of rEPO. Recently, different mutations in the noncoding region of the EPO gene were identified in patients with familial erythrocytosis. Their blood EPO profiles after IEF-PAGE were very basic and in the same area as rEPO. To avoid any possible misinterpretation in an antidoping context, the goal of this work was to characterize urine and blood EPO profiles from these patients using all methods authorized in antidoping laboratories (IEF-PAGE, SDS-PAGE, and SAR-PAGE) and to compare them with wild-type (WT) EPO and with profiles obtained after the administration of rEPO Eprex. Results showed that SDS-PAGE and SAR-PAGE profiles were not modified in the patients compared with WT EPO and could not be mistaken for the presence of rEPO. In contrast, the IEF-PAGE profiles in urine were slightly more acidic than in blood and closer to the rEPO profile. However, they did not meet the identification criteria used when this technique was the only authorized method. It is highly unlikely that these EPO mutations, only identified in erythrocytosis patients yet, could lead to an adverse analytical finding.
Analytical Challenges in Detecting New Psychoactive Substances in Anti-Doping: A Proof-of-Concept Involving Pentedrone Metabolite.
Kwiatkowska D, Peta B, Grucza K … +6 more , Łabęcka A, Konarski P, Wojtkowiak K, Grela K, Sytniczuk A, Wicka M
Drug Test Anal · 2026 Apr · PMID 41713878 · Full text
New psychoactive substances (NPS) have become increasingly prevalent worldwide, posing a growing challenge to public health, forensic science, and doping control. Their structural diversity, rapid emergence, and legal am...
New psychoactive substances (NPS) have become increasingly prevalent worldwide, posing a growing challenge to public health, forensic science, and doping control. Their structural diversity, rapid emergence, and legal ambiguity complicate both toxicological assessment and analytical detection. Research into the metabolism of these compounds is limited by ethical constraints on administration studies, hindering the development of reference standards. This study presents a methodological approach for the identification and confirmation of 2-amino-1-phenylpentan-1-ol, a metabolite associated with several synthetic cathinones, including pentedrone, α-pyrrolidinopentiophenone (PVP), and N-ethylpentedrone (NEP). The compound was detected in a routine doping control sample using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). In the absence of a commercially available certified reference material (CRM) encompassing all potential isomeric forms, a custom synthesis of the target metabolite was undertaken. The synthesized CRM-enabled unambiguous confirmation of the analyte and its diastereomeric composition, demonstrating the importance of reference compound availability for the accurate interpretation of results in anti-doping analysis. This work highlights both the analytical challenges posed by NPS and the need for continued development of tailored CRMs to support doping control laboratories worldwide.
Ethyl Glucuronide in Hair: A 10-Year Overview.
Casati S, Ravelli A, Bergamaschi RF … +6 more , Fabbri C, Palmisano E, Ostinelli A, Roda G, Facchi G, Orioli M
Drug Test Anal · 2026 Apr · PMID 41709682 · Full text
This 10-year retrospective study evaluates hair ethyl glucuronide (EtG), a direct metabolite of ethanol, in a large Northern Italian cohort (N = 68,221 samples collected between 2013 and 2022). The analysis aimed to eval...
This 10-year retrospective study evaluates hair ethyl glucuronide (EtG), a direct metabolite of ethanol, in a large Northern Italian cohort (N = 68,221 samples collected between 2013 and 2022). The analysis aimed to evaluate long-term alcohol consumption through the EtG distribution in terms of age, gender, recidivism, applicants, sampling region, sampling length, and the impact of seasonality and the COVID-19 pandemic. Hair EtG was determined by HPLC-MS/MS, and samples were classified according to the Society of Hair Testing (SoHT) recommended cut-offs: EtG ≤ 5 pg/mg (does not contradict abstinence), 5 < EtG < 30 pg/mg (repeated alcohol consumption), and EtG ≥ 30 pg/mg (chronic excessive alcohol consumption). A descriptive sensitivity analysis using SoHT cut-offs was performed to allow international comparability. The percentage (%) of hair samples classified as repeated or chronic excessive alcohol consumption was 13.6% (N = 9,281) and 5.6% (N = 3,818), respectively. EtG values significantly varied with gender and age as well as referral context. We observed statistically significant differences in EtG concentrations recorded in head, chest, axillary, and pubic hair samples. Significantly higher EtG values were detected in 3-cm proximal head hair versus the 3- to 6-cm proximal segment as well as across seasons, with a higher concentration in colder months. Conversely, no measurable short-term population impact of COVID-19 was revealed by hair EtG, whereas a significant long-term influence was highlighted. Overall, despite a large variability of EtG concentrations, this study provides robust evidence of the reliability of hair EtG testing and offers a comprehensive overview of long-term alcohol consumption trends in a monitored Italian population.
Identification and Analysis of Lysergic Acid Diethylamide Analogs, 4-Benzoyl-N,N-Diethyl-7-Methyl-4,6,6a,7,8,9-Hexahydroindolo[4,3-fg]quinoline-9-Carboxamide (1Bz-LSD) and N,N-Diethyl-7-Methyl-4-(4-(Trimethylsilyl)Benzoyl)-4,6,6a,7,8,9-Hexahydroindolo[4,3-fg]quinoline-9-Carboxamide (1-TMSBz-LSD), in tablet or paper sheet products available online in Japan.
Tanaka R, Kawamura M, Ito M … +1 more , Kikura-Hanajiri R
Drug Test Anal · 2026 Apr · PMID 41709646 · Publisher ↗
Recently, many lysergic acid diethylamide (LSD) analogs have emerged as designer drugs worldwide. In Japan, these compounds are distributed as paper sheet or tablet products and are available online. Even after these com...
Recently, many lysergic acid diethylamide (LSD) analogs have emerged as designer drugs worldwide. In Japan, these compounds are distributed as paper sheet or tablet products and are available online. Even after these compounds were regulated, new compounds with modified structures continued to appear. In the present study, two LSD analogs were identified from tablet or paper sheet products. Gas chromatography-mass spectrometry, liquid chromatography-photodiode array-mass spectrometry, liquid chromatography with hybrid quadrupole time-of-flight mass spectrometry, and nuclear magnetic resonance (NMR) measurement were used to determine the structures of the compounds. Based on these analyses, 4-benzoyl-N,N-diethyl-7-methyl-4,6,6a,7,8,9-hexahydroindolo[4,3-fg]quinoline-9-carboxamide (1Bz-LSD) and N,N-diethyl-7-methyl-4-(4-(trimethylsilyl)benzoyl)-4,6,6a,7,8,9-hexahydroindolo[4,3-fg]quinoline-9-carboxamide (1-TMSBz-LSD) were detected and identified in the tablet and paper sheet products, respectively. This paper is the first to report the detection of 1Bz-LSD and 1-TMSBz-LSD in tablet and paper sheet products in Japan.
Identification of Metabolites of the Dietary Supplement Ingredient 5α-Hydroxy-Laxogenin.
Derwand D, Zierau O, Thieme D … +1 more , Keiler AM
Drug Test Anal · 2026 Apr · PMID 41702844 · Full text
Illegally added, undeclared dietary supplement ingredients represent one way of unintentional exposure to substances that are prohibited by the World Anti-Doping Agency (WADA). In addition to the risk of anti-doping rule...
Illegally added, undeclared dietary supplement ingredients represent one way of unintentional exposure to substances that are prohibited by the World Anti-Doping Agency (WADA). In addition to the risk of anti-doping rule violation, adulterated dietary supplements can also be a health risk for elite and recreational athletes. 5α-hydroxy-laxogenin is one of those illegal substances but is not prohibited yet. Though an in vitro bioassay showed androgen receptor activation, this androgenic effect could not be confirmed in the preclinical orchiectomized rat model, which might be due to a fast biotransformation into inactive metabolites. In the present study, we investigated the metabolism of 5α-hydroxy-laxogenin using three different approaches: in silico prediction using the GLORYx tool of NERRD, in vitro biotransformation using the human hepatocellular cell line HepG2 and the preclinical orchiectomized rat model, from which serum and hair samples were collected and investigated. Analyses were performed using high-performance liquid chromatography-high-resolution mass spectrometry (HPLC-HRMS). In silico metabolism predicted five different metabolites resulting from hydroxylation, reduction, or oxidation. The human HepG2 cells generated five metabolites resulting from reduction or hydroxylation and from a combination of both. Three metabolites generated in vitro and an additional mono-hydroxylated metabolite were detected in the serum of the rats treated daily subcutaneously with 5α-hydroxy-laxogenin for 2 weeks. Hair analysis points to incorporation of 5α-hydroxy-laxogenin, though showing a rather high contamination most likely by saliva or urine. In conclusion, we identified and characterized metabolites of 5α-hydroxy-laxogenin, for which, however, the exact modification sites need to be investigated in future.
Annual Banned-Substance Review 18th Edition-Analytical Approaches in Human Sports Drug Testing 2024/2025.
Thevis M, Kuuranne T, Geyer H
Drug Test Anal · 2026 Apr · PMID 41702393 · Full text
Alongside the considerable advances and accomplishments in drug research and development, the breadth of anti-doping research topics has also continued to grow. This is particularly relevant not only to provide comprehen...
Alongside the considerable advances and accomplishments in drug research and development, the breadth of anti-doping research topics has also continued to grow. This is particularly relevant not only to provide comprehensive information on new drug entities, drug metabolism and elimination, and/or the impact of administration and exposure routes on analytical test results but also to ensure the timely implementation of analytical methods that ensure the availability of relevant anti-doping testing procedures and corresponding analytical data for routine doping control applications. The 18th edition of the annual banned-substance review on analytical approaches in human sports drug testing is dedicated to literature published between October 2024 and September 2025, and information published within these 12 months on established doping agents as well as new (potentially and evidently) relevant substances is reviewed and discussed, especially in the context of the World Anti-Doping Agency's 2025 Prohibited List. Topics of particular interest have been investigations into the metabolic fate and detection of anabolic agents, both anabolic-androgenic steroids and other anabolic substances such as selective androgen receptor modulators and protein-based therapeutics negatively regulating the activin receptor signaling pathway, and detection strategies for numerous new drug candidates have been discussed and presented. Further, the trend toward expanding testing options with regard to gene doping practices including oligonucleotide-based compounds (e.g., small interfering RNA, antisense oligonucleotides, etc.), transgenes, and gene editing practices continued also in 2024/2025, underlining the relevance for current and future sports drug testing programs.
Isomer Composition Assessment of Synthetic Phosphatidylethanol by Collision-Induced and Ozone-Induced Dissociation Mass Spectrometry.
Bantle M, Long JOT, Brydon SC … +4 more , Young RSE, Blanksby SJ, Weinmann W, Luginbühl M
Drug Test Anal · 2026 Apr · PMID 41702380 · Full text
The direct blood-alcohol biomarker phosphatidylethanol (PEth), especially its most abundant analogue 1-palmitoyl-2-oleoyl-sn-phosphatidylethanol (PEth 16:0/18:1) has gained increasing relevance in clinical and forensic a...
The direct blood-alcohol biomarker phosphatidylethanol (PEth), especially its most abundant analogue 1-palmitoyl-2-oleoyl-sn-phosphatidylethanol (PEth 16:0/18:1) has gained increasing relevance in clinical and forensic applications for assessing alcohol consumption. Accurate quantification of PEth is essential to reliably differentiate between abstinence, moderate alcohol consumption and excessive alcohol intake. Measurement accuracy of PEth 16:0/18:1 by well-established liquid chromatography-tandem mass spectrometry (LC-MS/MS) approaches such as multiple reaction monitoring (MRM) can be confounded by the presence of the regioisomer 1-oleoyl-2-palmitoyl-sn-phosphatidylethanol (PEth 18:1/16:0) in samples and synthetic reference standards. To address this measurement uncertainty, we conducted a new assessment of the isomeric composition of six currently available reference materials from four suppliers using collision-induced dissociation/ozone-induced dissociation (CID/OzID). Examination of these synthetic compounds found a high degree of regioisomeric purity of > 95%. Thus verified, the relative abundance of two key LC-MS/MS transitions were compared across a range of collision energies for both reference materials and an exemplary set of 10 dried blood spot case samples. These findings suggest a significantly wider range of natural isomer distributions spanning both higher and lower regiochemical composition (88.8%-98.85%) than the reference materials but within a range that would not significantly impact clinical classification.
In Vitro Metabolism of 1-Benzoyl-Lysergic Acid Diethylamide (1Bz-LSD) and Identification of a Deethylated Metabolite (1Bz-LAE) Using a Synthesized Reference Standard.
Azuma Y, Asada A, Tanaka M … +1 more , Doi T
Drug Test Anal · 2026 Apr · PMID 41700314 · Publisher ↗
Lysergic acid diethylamide (LSD) analogs represent an emerging class of new psychoactive substances (NPS). These compounds are often rapidly metabolized, making direct detection of the parent compound in biological sampl...
Lysergic acid diethylamide (LSD) analogs represent an emerging class of new psychoactive substances (NPS). These compounds are often rapidly metabolized, making direct detection of the parent compound in biological samples difficult. Therefore, metabolite detection serves as a critical and effective strategy for the confirmation of consumption. However, the metabolic pathways of these analogs remain largely uncharacterized, and synthesized metabolite standards for definitive identification are mostly unavailable. This study investigated the newly emerged LSD analog, (8R)-1-benzoyl-N,N-diethyl-6-methyl-9,10-didehydroergoline-8-carboxamide (1-benzoyl-LSD, 1Bz-LSD), which features a benzoyl moiety at the N position. The objectives were to characterize its metabolic fate and synthesize a targeted metabolite analyte for the confirmation of consumption. In vitro metabolism of 1Bz-LSD was examined using human liver microsomes and analyzed through liquid chromatography-quadrupole time-of-flight mass spectrometry. The parent compound was rapidly metabolized, yielding 15 metabolites, including LSD. Among these, a deethylated metabolite was detectable over a prolonged period, indicating its potential as a targeted analyte. To confirm its structure, this metabolite was chemically synthesized and identified as 1-benzoyl-lysergic acid ethylamide (1Bz-LAE). The synthesis and identification of 1Bz-LAE provide a crucial analytical foundation for the investigations of 1Bz-LSD use, with this metabolite anticipated to function as a valuable marker.
Analysis and Identification of In Vitro Metabolites of Exercise Mimetic SLU-PP-332 ERRα/β/γ Agonist for Doping-Control Purposes.
Avliyakulov NK, Sobolevsky T, Ahrens E
Drug Test Anal · 2026 Mar · PMID 41688415 · Publisher ↗
Exercise mimetics mimic physical activity and prevent development and progression of chronic metabolic diseases, including obesity and Type 2 diabetes. The World Anti-Doping Agency (WADA) prohibits the use of exercise mi...
Exercise mimetics mimic physical activity and prevent development and progression of chronic metabolic diseases, including obesity and Type 2 diabetes. The World Anti-Doping Agency (WADA) prohibits the use of exercise mimetics and metabolic modulators in sports. SLU-PP-332 is a small, synthetic ERRα/β/γ agonist recently developed using a rational drug design approach. SLU-PP-332 has been shown to increase oxidative muscle fibers, fatty acid oxidation, and enhance exercise endurance. In mouse models of metabolic syndrome, it increased energy expenditure and insulin sensitivity and conferred cardiac protection against pressure overload-induced heart failure by transcriptionally activating a wide spectrum of metabolic genes involved in fatty acid metabolism and mitochondrial function. The identification of metabolites of this emerging therapeutic molecule is a critical step towards detecting its misuse. The aim of the study was identification of Phase I and II metabolites generated in vitro using pooled human liver S9 fractions. Metabolites were analyzed using liquid chromatography-high-resolution mass spectrometry (LC-MS/HRMS). Five monohydroxylated (M1-M5), three dihydroxylated (M6-M8), and four reduced dihydroxylated metabolites (M9-M12) were identified. Metabolites M13 and M19 showed direct glucuronidation and sulfation of the parent compound, respectively. Metabolites M14-M18 and M20-M22 showed glucuronidation and sulfation with hydroxylation of the naphthalene or phenolic rings, respectively. M1, M7, M9, M10, M13, M14, M19, and M20 were the most abundant of the 22 metabolites and potentially useful for doping-control purposes. Further studies are necessary to fully elucidate the structures of the metabolites.
Urinary and Plasma Profiles of Betamethasone and Triamcinolone Acetonide Following Intra-Articular and Related Administrations.
Coll S, Bressan C, Monfort N … +6 more , Llorente-Onaindia J, Monfort J, Santiago J, Balius R, Brotons D, Ventura R
Drug Test Anal · 2026 Mar · PMID 41684241 · Full text
Since 2022, intra-articular (IA) and periarticular (PA) administrations of glucocorticoids (GC) have been prohibited in sports during competitions. The aim of this study was to evaluate plasma and urinary concentrations...
Since 2022, intra-articular (IA) and periarticular (PA) administrations of glucocorticoids (GC) have been prohibited in sports during competitions. The aim of this study was to evaluate plasma and urinary concentrations of betamethasone (BET) and triamcinolone acetonide (TA) following IA and PA administrations, in order to verify the suitability of the minimum reporting levels (MRL) and washout periods established by WADA and to assess their systemic effects. Urine and plasma samples were collected after IA or PA administration in patients or athletes receiving GC treatment. For BET, 12 participants received IA, and 20 received PA injections; for TA, eight participants received IA, and six received PA administrations. Sample analysis was performed using validated LC-MS/MS methods. The elimination profiles of BET after both IA and PA administrations were comparable, with high concentrations observed during the first postadministration day followed by a progressive decline. Similar plasma concentrations were obtained after both routes, with a similar effect on cortisol levels, indicating a clear systemic effect. For TA, different excretion patterns were observed depending on the administration route. After IA injections, TA displayed a profile similar to that of BET. In contrast, following PA administration, TA concentrations in urine and plasma were lower and remained relatively constant over several days. The degree and duration of cortisol suppression after PA administration varied among participants, with some showing a short effect. The results obtained support the MRL for IA and PA administrations defined by WADA. However, the washout period for BET is recommended to be increased to 5 days.
Distribution of Clomiphene and Its Metabolites in Antidoping Samples-A 3-Year Perspective.
Nair VS, Heybroek M, Miller GD … +3 more , Moore C, Campbell T, Eichner D
Drug Test Anal · 2026 Mar · PMID 41672565 · Publisher ↗
Clomiphene use substantially enhances endogenous testosterone production in males. Consequently, it is included under Section 4.2 of the World Anti-Doping Agency (WADA) Prohibited List and prohibited at all times (in and...
Clomiphene use substantially enhances endogenous testosterone production in males. Consequently, it is included under Section 4.2 of the World Anti-Doping Agency (WADA) Prohibited List and prohibited at all times (in and out of competition). Previous research has highlighted the prolonged excretion and detection windows for clomiphene and its metabolites after cessation of use. In addition to therapeutic use in humans, clomiphene may also be used to increase egg production in laying hens. Accordingly, clomiphene and/or its metabolites may also be detected in human urine after consumption of eggs or meat from treated poultry. To address some of these complexities, WADA has issued targeted guidance in Technical Letter TL26, mandating a minimum reporting level for clomiphene. This short communication evaluates results from clomiphene findings in 325 urine samples over the past 3 years and examines how reporting would be affected by the criteria outlined in TL26.
The 43rd Manfred Donike Workshop on Doping Analysis.
Thevis M
Drug Test Anal · 2026 Mar · PMID 41668325 · Publisher ↗
Abstract loading — click title to view on PubMed.
Emoxypine as a Potential Marker for Identifying Possible Cases of Unintended Meldonium Doping in Sports.
Postnikov PV, Efimova YA, Rodin IA
Drug Test Anal · 2026 Mar · PMID 41611212 · Publisher ↗
In recent years, a scenario involving milk contamination by biotransformation products of the veterinary drug Emidonol has received widespread publicity. Emidonol is intended for use in cattle under pathological conditio...
In recent years, a scenario involving milk contamination by biotransformation products of the veterinary drug Emidonol has received widespread publicity. Emidonol is intended for use in cattle under pathological conditions accompanied by hypoxia. The drug dissociates into two antihypoxants-meldonium, a metabolic modulator prohibited in sports, and emoxypine, which also has antioxidant properties. A small-scale pilot study involved three volunteers consuming a single 900-mL dose of unpasteurized cow's milk collected on the last (15th) day of treatment with the drug. The mean estimated urinary concentration of emoxypine peaked between 7- and 8.5-h postadministration (1360 ± 240 ng/mL), elimination time being approximately 50-54 h. The mean estimated urinary concentration of meldonium peaked between 5.8- and 9.0-h postadministration and was 322 ± 68 ng/mL, elimination time being approximately 35-45 h. The simultaneous presence of emoxypine and meldonium in urine samples may be indicative of the consumption of Emidonol-contaminated milk, which may facilitate the identification of cases of unintended doping in sport.
Lomerizine M6-An Important Urinary Metabolite in Anti-Doping Control Analysis for Correct Detection of Trimetazidine Abuse in Both Females and Males.
Bohlin KP, Ekström L, Pohanka A … +2 more , Lehtihet M, Ericsson M
Drug Test Anal · 2026 Mar · PMID 41582662 · Publisher ↗
Distinguishing between intake of the prohibited substance trimetazidine from administration of approved migraine medicine containing the nonprohibited substance, lomerizine, is crucial for anti-doping control laboratorie...
Distinguishing between intake of the prohibited substance trimetazidine from administration of approved migraine medicine containing the nonprohibited substance, lomerizine, is crucial for anti-doping control laboratories. Investigations in males after lomerizine intake have been conducted leaving lomerizine M6 as the most useful metabolite. To our knowledge, the excretion profile of lomerizine among women has not been studied and leaves a potential gap to be misused by athletes in the case of appeal of a reported adverse analytical finding. We have investigated lomerizine M6 in women to address the gender-specific biology to correctly detect misusage of the prohibited substance trimetazidine within sports. To determine the importance of lomerizine M6 in trimetazidine cases, we analyzed urine samples collected from 12 individuals (eight females and four males) over 144 h, after oral intake of 5-mg lomerizine dihydrochloride. A dilute and shoot method was used, and the samples were analyzed on a LC-HRMS instrument. In all study subjects, trimetazidine was found from 2 h up to 48 h, and lomerizine M6 was detected from 2 h and onward. Lomerizine was only detected above the limit of identification of the method for a few of the subjects. The study demonstrated that by monitoring lomerizine M6, it was possible to determine the origin of trimetazidine in women. The metabolism for lomerizine dihydrochloride does not appear to differ significantly between genders. However, lomerizine M6 concentrations are not always higher than trimetazidine concentrations. Special care should be taken when interpreting trimetazidine concentrations at 1 ng/mL or below.
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