{"title":"Origin and Interpretation of Low Methamphetamine Levels Found in Amphetamine-Positive Urine Samples: Support for Methylation of Amphetamine as a Minor Metabolic Pathway.","authors":"Anders Helander, Annika Andersson, Tomas Villén","doi":"10.1002/dta.3940","DOIUrl":"https://doi.org/10.1002/dta.3940","url":null,"abstract":"<p><p>Methamphetamine is relatively uncommon on the European drug market, but Swedish drug test laboratories repeatedly detect low methamphetamine concentrations in urine samples containing high amphetamine levels, warranting clinical evaluation for suspected polydrug use. Of 12,062 routine samples screened positive for amphetamines, 86% were confirmed positive (≥ 200 μg/L) for amphetamine, 2.1% for methamphetamine, and 4.0% for 3,4-methylenedioxymethamphetamine (ecstasy) by liquid chromatography-tandem mass spectrometry. Of the 259 methamphetamine-positive samples, 98% contained amphetamine concentrations above the reporting limit, consistent with the metabolic conversion of methamphetamine to amphetamine in the body. However, in most (69%) of these samples, the methamphetamine concentration was only < 2% of the amphetamine level, suggesting methamphetamine was not the primary drug taken. Chiral analysis of selected samples showed that after use of illicit street amphetamine with a racemic content of the l- and d-enantiomers, a similar l/d proportion was observed for methamphetamine. Similarly, in samples from patients receiving d-amphetamine-based ADHD medication, low d-methamphetamine levels were detected, even though the pharmaceutical products contained no methamphetamine. This observation, together with the parallel l/d-enantiomer distributions, supports amphetamine N-methylation as a trace, albeit quantitatively insignificant, metabolic pathway in humans. From both a clinical and forensic perspective, a low urinary methamphetamine concentration of less than a few percent of the amphetamine level therefore does not warrant further clinical evaluation for suspected polydrug use. The present findings further demonstrate that chiral analysis of both amphetamine and methamphetamine is an effective approach for distinguishing between illicit and therapeutic sources in positive screening drug tests for the amphetamines.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144797739","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Gregory A Brown, Brandon Shaw, Ina Shaw, Michael J Joyner, Sandra K Hunter, Jonathon W Senefeld, Ross Tucker, Emma Hilton, Tommy R Lundberg, Chad Carlson, Christopher Kirk
{"title":"Comment on \"Biology and Management of Male-Bodied Athletes in Elite Female Sports\" by Handelsman and Bermon.","authors":"Gregory A Brown, Brandon Shaw, Ina Shaw, Michael J Joyner, Sandra K Hunter, Jonathon W Senefeld, Ross Tucker, Emma Hilton, Tommy R Lundberg, Chad Carlson, Christopher Kirk","doi":"10.1002/dta.3916","DOIUrl":"https://doi.org/10.1002/dta.3916","url":null,"abstract":"","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144797738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Response to Letter to Editor From Brown et al.","authors":"D J Handelsman, S Bermon","doi":"10.1002/dta.3917","DOIUrl":"https://doi.org/10.1002/dta.3917","url":null,"abstract":"","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144797740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Catalina Dumitrascu, Elias Iturrospe, Els Scheir, Patrick Timmermans, Erik Fransen, Matthias Van Puymbroeck, Glenn Van Nieuwenhove, Maryline Busschots, Diona D'Hondt, Alexia Van Goethem, Wout Claeys, Babette Van Rafelghem, Eline Baetens, Philippe G Jorens, Celine Gys, Werner Jacobs, Hugo Neels, Adrian Covaci, Alexander L N van Nuijs
{"title":"Ethylated Phosphorylcholine as a New Marker for Alcohol Consumption: A Proof of Concept.","authors":"Catalina Dumitrascu, Elias Iturrospe, Els Scheir, Patrick Timmermans, Erik Fransen, Matthias Van Puymbroeck, Glenn Van Nieuwenhove, Maryline Busschots, Diona D'Hondt, Alexia Van Goethem, Wout Claeys, Babette Van Rafelghem, Eline Baetens, Philippe G Jorens, Celine Gys, Werner Jacobs, Hugo Neels, Adrian Covaci, Alexander L N van Nuijs","doi":"10.1002/dta.3937","DOIUrl":"https://doi.org/10.1002/dta.3937","url":null,"abstract":"<p><p>Alcohol consumption is widespread worldwide and a leading cause of injuries, morbidity, and mortality. Accurately detecting alcohol use with reliable biomarkers is crucial in clinical and forensic settings. Direct alcohol biomarkers, i.e., ethanol (EtOH), ethyl glucuronide (EtG), ethyl sulphate (EtS), phosphatidylethanol 16:0/18:1 (PEth) reflect short- and long-term consumption. Nevertheless, complementary biomarkers with improved specificity and sensitivity are needed to better assess alcohol use, including generating a detailed timeline of consumption. In vitro exposure of HepaRG liver cells to EtOH resulted in the generation of ethylated phosphorylcholine (EtOChoP). This is the first study to investigate the in vivo presence of EtOChoP and its occurrence in medico-legal samples. Proof-of-concept and observational studies assessed EtOChoP, PEth, EtG, EtS, and EtOH in whole blood, and, when available, other matrices were analyzed for EtG, EtS (plasma, serum, urine, hair), EtOH (urine), and EtOChoP (plasma, serum). A single alcohol exposure event (0.5 g/kg EtOH, with blood EtOH concentration peaking at 0.76 g/L at 100 min) led to EtOChoP presence, and, similar to short-term biomarkers (e.g., EtOH, EtG, and EtS in whole blood), EtOChoP was not detected in the following day(s). However, in the observational study, EtOChoP remained detectable even when short-term biomarkers were absent, resembling long-term biomarkers (PEth and hair EtG). Notably, 14% of samples were positive only for EtOChoP, highlighting the need for additional biomarkers. These findings identify EtOChoP as a promising alcohol (ab)use biomarker formed after EtOH consumption and possibly accumulating during chronic drinking. EtOChoP could potentially differentiate between recent drinking and chronic problematic drinking in individuals with high PEth levels.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Olga V Kupriyanova, Vadim A Shevyrin, Raziya G Sadykova, Yuri M Shafran
{"title":"Differentiating Positional Isomers of the 2-(Dimethoxyphenyl)-N-(2-(trifluoromethoxy)benzyl)ethanamine Series: A Chromatography-Mass Spectrometry Approach.","authors":"Olga V Kupriyanova, Vadim A Shevyrin, Raziya G Sadykova, Yuri M Shafran","doi":"10.1002/dta.3936","DOIUrl":"https://doi.org/10.1002/dta.3936","url":null,"abstract":"<p><p>Synthetic derivatives of phenylethanamine, specifically 2-(2,5-dimethoxyphenyl)-N-((2-substituted)benzyl)ethanamines, regularly appear on the global market of new designer drugs and have been reported to be associated with adverse effects in humans who consume them. At the same time, their positional isomers may lack psychoactive effects. These compounds are not currently regulated by legislation and could potentially be used for medical purposes in the future. Continuing the study of the properties of 2-(2,5-dimethoxyphenyl)-N-((2-substituted)benzyl)ethanamines with different substituents, a poorly studied series of 2-(dimethoxyphenyl)-N-(2-(trifluoromethoxy)benzyl)ethanamines (NBOMe(F)) with a trifluoromethoxy group (OCF<sub>3</sub>) in the ortho-position of the N-benzyl fragment is of interest. Gas chromatographic analysis revealed the existence of a critical pair of isomers when using an HP-5 type column, the differentiation of which is difficult under varying temperature conditions of the column thermostat. Their separation was achieved on a DB-17MS column under isothermal conditions at a temperature of 210°C. A method using thin-layer chromatography for the differentiation of this critical pair of isomers is also proposed. Retention indices of the isomeric compounds of the NBOMe(F) series and their N-trifluoroacetyl derivatives have been determined, which can serve as an additional identification criterion for gas chromatographic analysis. It has been shown that differentiation of NBOMe(F) isomers using electron ionization mass spectra is only possible if the spectra of both the isomers themselves and their N-trifluoroacetyl derivatives, which exhibit significant differences, are recorded. As an alternative method for differentiating the isomeric compounds, an approach using high-performance liquid chromatography-tandem mass spectrometry, which does not require derivatization, is proposed.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144937300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Max Polke, Florian Zapf, Tanja Restin, Thomas Kraemer, Tina M Binz
{"title":"Evaluation of Therapeutic Opioids in Hair of Neonatal and Pediatric Patients.","authors":"Max Polke, Florian Zapf, Tanja Restin, Thomas Kraemer, Tina M Binz","doi":"10.1002/dta.3935","DOIUrl":"https://doi.org/10.1002/dta.3935","url":null,"abstract":"<p><p>Forensic hair analysis poses a valuable tool for assessing opioid exposure in children and neonates. However, reliable literature data on opioid concentrations in the hair of this population are mostly scarce, making the interpretation of such hair analysis results rather challenging. This noninterventional study aims to address this issue by investigating 118 hair samples of pediatric patients (median age: 50 days) from the University Children's Hospital Zurich. These patients were treated with medically approved novel synthetic opioids (fentanyl, remifentanil, sufentanil, or alfentanil) and traditional opioids (morphine, methadone, and hydromorphone) during their clinical treatment. Quantification of the opioids and selected metabolites was achieved by a previously validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) based method, which showed good sensitivity with lower limits of quantification (LLOQs) ranging from 0.1 to 1 pg/mg hair. Most analytes were successfully detected in patients' hair, with the majority being identified for the first time in this matrix. Significant correlations were found between the opioid concentrations in hair and the administered medication doses, indicating that hair analysis may reflect the extent of opioid exposure in this population. Furthermore, metabolite ratios similar to the ones commonly found in adult hair were identified, which are forensically important to differentiate between active intake of a drug and contamination. The metabolite ratio of β-hydroxyfentanyl to fentanyl was particularly well suited for children and neonatal patients. In conclusion, concentration ranges, metabolite ratios, and dose correlations of the studied opioids in pediatric hair were established, providing insights into opioid incorporation pathways.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144726240","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sara Casati, Alessandro Ravelli, Roberta F Bergamaschi, Massimo Del Fabbro, Giorgio Binelli, Gabriella Roda, Marica Orioli
{"title":"Long-Term Stability of Ethyl Glucuronide in Hair: A 10-Year Retrospective Analysis of 909 Samples by LC-MS/MS.","authors":"Sara Casati, Alessandro Ravelli, Roberta F Bergamaschi, Massimo Del Fabbro, Giorgio Binelli, Gabriella Roda, Marica Orioli","doi":"10.1002/dta.3934","DOIUrl":"https://doi.org/10.1002/dta.3934","url":null,"abstract":"<p><p>Monitoring long-term alcohol consumption is critical in forensic and public health contexts. Hair analysis of ethyl glucuronide (EtG), a direct metabolite of ethanol, has become a standard method for detecting chronic alcohol use. While the reliability of EtG hair testing is well established for short- and medium-term analyses, its stability in hair stored over extended periods has not been comprehensively evaluated. This limitation is especially relevant in retrospective investigations, postmortem evaluations, and long-term epidemiological studies, where archived samples may be analyzed years after collection. In this study, we assessed the long-term stability of EtG in human hair stored for up to 10 years. A total of 909 samples originally analyzed between 2013 and 2022 were re-tested in 2023 using a previously published and validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. When the results of the old and the new analyses were compared, EtG concentrations showed no significant degradation over time, with more than 80% of the samples displaying matching values when analytical uncertainty was considered. Only a small fraction of samples (4.4%) dropped below the commonly used interpretive threshold for chronic alcohol use (30 pg/mg) after 10 years of storage. These findings provide robust evidence that EtG remains chemically stable in hair under standard storage conditions over a decade, confirming the reliability of archived samples for assessing alcohol use history and expanding the utility of EtG analysis in long-term toxicological and forensic investigations. The demonstrated stability strengthens confidence in hair as a matrix for retrospective substance use evaluation across scientific disciplines.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144688473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Evaluation of Two Spectroscopic Techniques to Estimate the MDMA Dose of Ecstasy-Like Tablets, an On-Site Approach.","authors":"N Meert, K Segers, F Van Durme, J Eliaerts","doi":"10.1002/dta.3933","DOIUrl":"https://doi.org/10.1002/dta.3933","url":null,"abstract":"<p><p>MDMA, commonly known as \"ecstasy,\" is widely used in clubs and at festivals, earning its reputation as a \"party drug.\" The increasing demand for rapid on-site dose estimation of MDMA in tablets arises from the need of various stakeholders, including law enforcement, emergency services, and public health officials, to respond appropriately to potential public safety risks and incidents. This study evaluates the performance of two portable spectroscopic techniques (near-infrared [NIR] and Fourier-transform-infrared [FT-IR]) combined with chemometric modelling for estimating the MDMA dose in tablets. Ninety-eight seized tablets were measured on-site with both spectroscopic techniques and confirmed by the reference techniques: gas chromatography (GC) combined with a mass spectrometer (MS) and a flame-ionization detector (FID). Considering the correlation values (NIR: R<sup>2</sup> = 0.64 for indirect contact with intact tablets, 0.87 for direct contact with homogenized tablets; FT-IR: R<sup>2</sup> = 0.84) and the RMSEP values (NIR: 8.0 for indirect contact with intact tablets, 5.9 for direct contact with homogenized tablets; FT-IR: 5.4), both spectroscopic techniques provide a reliable dose prediction in comparison to the GC-FID results. Moreover, these devices are suitable for rapid on-site application. The instruments' choice depends on several factors, such as speed, safety measures, and laboratory support for modelling. However, determining the MDMA dose does not address all health risks. Other factors, such as the presence of low-dosed substances (undetectable on-site) and the combination of MDMA with other drugs and/or alcohol also play a significant role. Therefore, laboratory analysis remains essential for comprehensive safety assessment.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144688472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Carmel Heiland, Oscar Hopcraft, Mats Johanson, Anton Pohanka, Mikael Lehtihet, Lena Ekström
{"title":"Detection of a Single Microdose of Testosterone and Recombinant EPO in Healthy Volunteers.","authors":"Carmel Heiland, Oscar Hopcraft, Mats Johanson, Anton Pohanka, Mikael Lehtihet, Lena Ekström","doi":"10.1002/dta.3932","DOIUrl":"https://doi.org/10.1002/dta.3932","url":null,"abstract":"<p><p>Doping with testosterone (T) and erythropoietin (EPO) in low doses (micro-doping) is a challenge to detect. Here, we have investigated the ability to detect micro-doping of recombinant human (rhEPO) and testosterone (T) after administration of a single dose of subcutaneous Eporatio (15 IU/kg) and transdermal Testogel (100 mg) to healthy males. For rhEPO detection in urine, MAIIA EPO purification kits 3F6 (#1410) and 7D3 (#1460) were used for ITP and CP analyses, respectively, whereas kit 3F6 (#1430) was used for dried blood spots (Tasso). The sensitivity to detect rhEPO in Tasso was investigated and compared with previous detection results for Capitainer and Mitra. For T detection, the urinary and capillary serum steroid profile and IRMS analysis were performed. It was possible to detect administration of 15 IU/kg Eporatio with high sensitivity with our ITP method up to 72 h after administration, and the CP findings supported the ITP findings. Tasso provides less sensitivity in detecting Eporatio than Mitra and Capitainer. With IRMS, 100% of the samples analyzed were positive, also when not associated with elevated urinary T/E or 5αAdiol/E ratios. As an alternative, high systemic T levels aligned with positive IRMS results, highlighting the value of serum T as a complementary biomarker. Overall, the World Anti-Doping Agency ITP and CP methods employed today show good sensitivity towards detection of micro-dosing of rhEPO and T.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144681762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lu Liu, Zhanliang Wang, Lingyu Zhao, Xinmiao Zhou, Lisi Zhang
{"title":"Method Development of Pegmolesatide for Doping Analysis: A Novel Synthetic Erythropoietin-Mimetic Agent.","authors":"Lu Liu, Zhanliang Wang, Lingyu Zhao, Xinmiao Zhou, Lisi Zhang","doi":"10.1002/dta.3931","DOIUrl":"https://doi.org/10.1002/dta.3931","url":null,"abstract":"<p><p>Pegmolesatide, a novel synthetic erythropoietin-mimetic agent, was developed by the Hansoh Pharmaceutical Manufacturing Company Ltd. (Jiangsu, China). In late 2023, it was approved in China for the treatment of anemia in both dialysis and non-dialysis patients with chronic kidney disease, with the advantages of reduced immunogenicity and extended duration of action. The aim of this study was to develop a strategy for detecting pegmolesatide in doping analysis. Here, we present a bottom-up nano-liquid chromatography-high-resolution tandem mass spectrometry approach for qualitative analysis of pegmolesatide using erythropoietin receptor coupled magnetic beads, followed by trypsinization and subsequent detection of characteristic peptides. Using full scan and data-dependent MS/MS (ddMS2) modes, two characteristic peptide segments of pegmolesatide were identified. An analytical method using product ion scan mode was developed to detect the identified characteristic peptides. Both peptide segments were analyzed for the initial testing procedure, whereas one characteristic peptide segment obtained from complete trypsinization was analyzed for the confirmation procedure. After full validation, the selectivity, reliability, limit of detection, limit of identification, carryover, and stability were evaluated. The results demonstrate that our developed method can be a fit-for-purpose analytical method for the antidoping of pegmolesatide.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144657904","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}