Drug Test Anal [JOURNAL]
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200 papers
A Multitarget Screening Method for the Detection of Small Peptides in Dried Blood Spots for Doping Control Analysis.
Boschetti G, Langer T, Medana C … +6 more , Grabherr S, Kuuranne T, Salamin O, Nicoli R, Mumenthaler C, Musenga A
Drug Test Anal · 2026 Mar · PMID 41579148 · Publisher ↗
Over the past few years, dried blood spots (DBS) have been approved as a valid matrix for drug testing in sport. They undoubtedly offer advantages but also pose analytical challenges. For example, various DBS supports ar...
Over the past few years, dried blood spots (DBS) have been approved as a valid matrix for drug testing in sport. They undoubtedly offer advantages but also pose analytical challenges. For example, various DBS supports are nowadays available (polymeric and cellulose based) and an extraction method suitable for one support is not necessarily transferable to a different one. Herein, we present a qualitative screening method for the detection of a representative selection of small peptides and their metabolites in both cellulose and polymeric DBS. The analytes were extracted using an extraction solvent containing formic acid 1% in water/acetonitrile/methanol (70/15/15), followed by a second extraction with acetate buffer. To remove interferences and increase sensitivity, the combined extracts were further purified using solid-phase extraction (mixed-mode, weak cation exchange). Analysis was performed using ultrahigh-performance liquid chromatography combined with high-resolution mass spectrometry (orbitrap Q-Exactive). The analysis time was 7.5 min, and the acquisition was performed in full-scan mode, with the addition of some product ion scan acquisitions to increase selectivity or sensitivity for a few compounds that were particularly challenging. The method permits the analysis of small peptides on both polymeric and cellulose DBS samples with the same procedure for either matrix. Validation was performed following the World Anti-Doping Agency regulations, and the method proved satisfactory in terms of selectivity and sensitivity (limits of detection in the low ng/mL range) and applicable to the analysis of sport samples for the detection of small peptides.
Application of Novel Patent-Based Effervescent Tablet as a Selective Filter for Removing Hazardous Chemical Materials in Hookah Smoke: Determination and Comparison of Toxic Chemical Compounds in Hookah Smoke Versus Cigarette Smoke.
Raoufi A
Drug Test Anal · 2026 Mar · PMID 41558994 · Publisher ↗
A novel patent-based effervescent tablet was developed and evaluated as a selective filter for reducing hazardous chemical compounds in hookah smoke. The study combined gas chromatography-mass spectrometry (GC-MS) and at...
A novel patent-based effervescent tablet was developed and evaluated as a selective filter for reducing hazardous chemical compounds in hookah smoke. The study combined gas chromatography-mass spectrometry (GC-MS) and atomic absorption spectroscopy (AAS) to identify and quantify major toxicants. Hookah smoke was trapped using cellulose nanofiber-based solvent chambers and latex puff chambers, followed by analysis of volatile organic compounds (nicotine, propylene oxide, formaldehyde, crotonaldehyde, acrolein, acetaldehyde, 1,3-butadiene, benzopyrene, pyridine, phenol, benzene) and potentially toxic trace elements (PTEs) such as lead, mercury, cadmium, and chromium. Detection limits (LODs) ranged from 0.003 to 0.04 mg/g, with relative standard deviations (RSDs) below 13%. The effervescent tablet reduced > 81% of hazardous compounds while selectively lowering nicotine and flavor by approximately 15%, maintaining user acceptability. Compared with cigarette smoke, unfiltered hookah smoke contained significantly higher levels of aldehydes and polycyclic aromatic hydrocarbons, whereas filtered hookah smoke showed markedly lower contaminant concentrations than both cigarette and e-cigarette smoke. The proposed method demonstrates innovation by integrating a user-friendly, selective effervescent tablet with high recovery (92%-102%) and accuracy, offering a practical strategy for harm reduction in hookah smoking.
Detection and Longitudinal Monitoring of Cyclosporine in Horse Plasma and Urine After Ocular Implant Administration.
So YM, Kwok WH, Yap C … +5 more , Wong COL, Smalley SGR, Forbes BS, Chow DWY, Ho ENM
Drug Test Anal · 2026 Mar · PMID 41549061 · Publisher ↗
This paper describes the detection and longitudinal monitoring of cyclosporine in plasma and urine after subconjunctival implant administration in a horse. Sensitive liquid chromatography tandem mass spectroscopy (LC-MS/...
This paper describes the detection and longitudinal monitoring of cyclosporine in plasma and urine after subconjunctival implant administration in a horse. Sensitive liquid chromatography tandem mass spectroscopy (LC-MS/MS) methods for detecting cyclosporine in horse plasma and urine have been developed and validated, with estimated limits of detection down to 1 pg/mL in both matrices. The developed methods enabled longitudinal monitoring of cyclosporine levels in blood and urine samples collected over 6 months from a horse that had received an ocular cyclosporine implant.
A Robust Multistep Digestion Method for Microplastics Detection in Human Tissue by MicroRaman Analysis.
Pascali JP, Litti L, Fornasari A … +5 more , Giorgetti A, Pozzebon M, Barone R, Ragusa A, Fais P
Drug Test Anal · 2026 Mar · PMID 41500815 · Full text
The presence of microplastics (MPs) in human tissues has raised growing concerns, necessitating robust protocols for their reliable extraction and analysis. This study systematically evaluated and optimized digestion pro...
The presence of microplastics (MPs) in human tissues has raised growing concerns, necessitating robust protocols for their reliable extraction and analysis. This study systematically evaluated and optimized digestion protocols to efficiently process a variety of human tissues-placenta, lung, kidney, adipose tissue, muscle, spleen, liver, thyroid, and brain-while preserving the integrity of MP particles. Initial assessments employing single-reagent protocols such as nitric acid (HNO), proteinase K enzymatic digestion, and Fenton oxidative digestion demonstrated limited effectiveness, due to incomplete tissue breakdown or formation of turbid digestates that hindered filtration. Building upon these results, combined digestion approaches were investigated to improve organic matter removal and facilitate filtration through fine pore-size filters (0.2 μm). The optimized 3-day protocol included an initial oxidative Fenton digestion followed by enzymatic digestion (proteinase K). The final step involved lipid removal through ethanol addition and sonication, resulting in clear digestates amenable to filtration. This protocol efficiently digested complex tissue matrices, reducing filter clogging at 1-μm size pore and preserving various common MP polymers, including low-density polyethylene (LDPE), polyethylene terephthalate (PET), polytetrafluoroethylene (PTFE), and polyamides (PA6 and PA12). Application of the optimized digestion allowed successful isolation and characterization of MPs using optical microscopy and Raman spectroscopy. The method showed improved reproducibility and reliability over single-reagent protocols, making it suitable for comprehensive MP analysis in human tissues. The application of an efficient and robust protocol for tissue digestion may contribute to advance human exposure assessment and toxicological studies related to MP contamination.
Pesticide Contamination in the Hair of Children From Colonia San Juan, a Rural Community in Paraguay.
Benitez Leite S, Iglesias-Gonzalez A, Mesquita MN … +3 more , Macchi ML, Mesnage R, Appenzeller BMR
Drug Test Anal · 2026 Mar · PMID 41454582 · Full text
Chronic exposure to pesticides can cause carcinogenic, reproductive, neurological, and endocrine-disrupting effects. Hair analysis is a valuable biomonitoring tool to assess human exposure to pesticides. We determined th...
Chronic exposure to pesticides can cause carcinogenic, reproductive, neurological, and endocrine-disrupting effects. Hair analysis is a valuable biomonitoring tool to assess human exposure to pesticides. We determined the presence of pesticides, their metabolites, and other environmental pollutants in the hair of children in an agricultural area of Paraguay. We analyzed 152 pesticides and environmental chemicals in hair samples from 51 children (2-14 years, mean ± SD = 8.5 ± 3.3 years) living in Colonia San Juan, a rural community in Paraguay. The locality is surrounded by soybean crops, and the community engages primarily in family farming. Eighty of the 152 compounds (52.6%) were detected. Each child's sample contained an average of 55 ± 3.7 compounds (range 48-65), including organophosphates, pyrethroids, neonicotinoids, fungicides, herbicides, and endocrine disruptors such as bisphenol A and bisphenol S. Thirty-seven compounds were present in all samples. Children in this rural community are simultaneously exposed to numerous pesticides and pollutants, highlighting the urgent need for stricter environmental protections and preventive health measures.
Investigations Into the Human Metabolism of Trestolone (7α-Methyl-19-Nortestosterone).
Piper T, Fusshöller G, Thevis M
Drug Test Anal · 2026 Mar · PMID 41407450 · Full text
Already in the 1960s, the anabolic properties of Trestolone (7α-methyl-19-nortestosterone, MENT) were investigated in the context of cancer research, and MENT was found to be 10 times more potent regarding its anabolic p...
Already in the 1960s, the anabolic properties of Trestolone (7α-methyl-19-nortestosterone, MENT) were investigated in the context of cancer research, and MENT was found to be 10 times more potent regarding its anabolic properties compared to testosterone. The human metabolism of MENT was only investigated once in an antidoping context, and three urinary metabolites were identified, corroborating earlier findings from in vitro and animal experiments. Based on these metabolites, no doping control sample was reported to contain MENT or its metabolites in the last two decades albeit MENT is readily available via online distributors. One reason for the lack of adverse analytical findings in doping controls could be analytical challenges originating from the chromatographic properties of MENT and its urinary metabolites. Therefore, the human metabolism of MENT was reinvestigated employing an excretion study with deuterated MENT and metabolite detection based on hydrogen isotope ratio mass spectrometry in combination with high accuracy/high resolution mass spectrometry. Considering unconjugated, glucuronidated, and sulfated metabolites, 50 potential candidates were detected. In order to identify those metabolites suitable for sports drug testing, three volunteers administered a single oral dose of nondeuterated MENT, and all postadministration samples were investigated using triple quadrupole mass spectrometry-based determinations routinely employed in doping controls. From the 50 metabolites detected, two showed promising results with respect to their detection windows and suitability under routine measurement conditions. The specificity of the novel metabolites was ensured by the reanalysis of 200 routine doping control samples demonstrating the absence of potential coeluting compounds.
Identification and Structural Elucidation of a New Synthetic Cannabinoid, MDMB-5'Br-PINACA, in Seized Herbal Materials.
de Godoi AB, Zeoly LA, Lanaro R … +4 more , Oliveira IS, Diana MC, Yonamine M, Costa JL
Drug Test Anal · 2026 Mar · PMID 41403075 · Full text
Synthetic cannabinoids (SCs) remain the most frequently detected class of new psychoactive substances (NPS) worldwide. Despite a recent decline in the overall number of newly reported NPS, SCs continue to emerge with rem...
Synthetic cannabinoids (SCs) remain the most frequently detected class of new psychoactive substances (NPS) worldwide. Despite a recent decline in the overall number of newly reported NPS, SCs continue to emerge with remarkable structural diversity. Here, we report the discovery and structural elucidation of MDMB-5'Br-PINACA, a previously unreported SC identified in three seized herbal materials. The compound was isolated by semipreparative liquid chromatography and subsequently characterized using an integrated analytical approach combining gas chromatography-mass spectrometry (GC-MS), liquid chromatography-high-resolution mass spectrometry (LC-HRMS), and nuclear magnetic resonance (NMR) spectroscopy. In addition to MDMB-5'Br-PINACA, other SCs were detected in the analyzed materials, such as 5F-ADB and MDMB-4en-PINACA, also including the synthetic precursor MDMB-INACA. Other NPS classes were also observed, including designer benzodiazepines (N-desalkylgidazepam and bromazolam), and synthetic opioids (metonitazene). Recent years have also seen the emergence of brominated SCs as a strategy to evade legislative control, with several 5-bromo analogs detected across different regions. This analytical workflow enabled the unambiguous identification of MDMB-5'Br-PINACA and provided a detailed chemical profile of the seized samples, highlighting the continued evolution and complexity of NPS mixtures in herbal formulations. The findings emphasize the importance of continuous monitoring and early detection of emerging substances, which are essential not only for forensic and toxicological investigations but also for public health surveillance and the development of evidence-based drug control and harm-reduction policies.
27th Meeting of the Society of Hair Testing (SoHT).
Favretto D, Rossi SS, Appenzeller B … +1 more , Cirimele V
Drug Test Anal · 2026 Mar · PMID 41388672 · Publisher ↗
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New Metabolites of Methyldienolone by In Vitro Human Liver Microsomes and Characterized Using LC/HRMS for Doping Control Purposes.
Hou J, Yan X, Zhang L … +2 more , Wang Z, Yang S
Drug Test Anal · 2026 Mar · PMID 41387186 · Publisher ↗
Methyldienolone, a synthetic anabolic androgenic steroid (AAS), has been banned in sports by the World Anti-Doping Agency (WADA) because of its performance-enhancing properties. This study aimed to investigate the main m...
Methyldienolone, a synthetic anabolic androgenic steroid (AAS), has been banned in sports by the World Anti-Doping Agency (WADA) because of its performance-enhancing properties. This study aimed to investigate the main metabolites using in vitro incubation with human liver microsomes (HLM) and to detect them through liquid chromatography-high-resolution mass spectrometry (LC-HRMS) for doping control purposes. A total of six groups of Phase I metabolites, including 17-epimerization, hydroxylation, C3-keto reduction, 18-nor modifications, reduction, and demethylation, as well as five different Phase II metabolites, such as glucuronide conjugates, were characterized, indicating extensive metabolism by HLM. Structural characterization of these metabolites was improved through derivatization with methoxylamine and hydroxylamine, which enabled their detection with higher sensitivity by LC-HRMS. These novel metabolites provide new insights into the metabolism of methyldienolone and may contribute to antidoping analysis. The synthesis of reference materials is necessary to confirm the structure of the proposed metabolites in the future.
A First Report of the Misuse of a Novel Synthetic Glucocorticoid, 9α-Fluoro-6α-Methylprednisolone in Camel Racing.
Ishii H, Basgallop NJ, Kelly RP … +3 more , Todoroki K, Alshamsi NS, McKinney AR
Drug Test Anal · 2026 Feb · PMID 41386684 · Publisher ↗
We report the misuse of a novel synthetic glucocorticoid, 9α-fluoro-6α-methylprednisolone (9F6MP) for the first time in camel racing and, to the best of our knowledge, human or other animal sports. During routine post-ra...
We report the misuse of a novel synthetic glucocorticoid, 9α-fluoro-6α-methylprednisolone (9F6MP) for the first time in camel racing and, to the best of our knowledge, human or other animal sports. During routine post-race drug testing of cameline plasma samples using liquid chromatography-tandem mass spectrometry we encountered an unknown peak with the same selected reaction monitoring traces as a dexamethasone formate adduct but at a different retention time. The product ion mass spectrum of the unknown peak in negative ion mode was identical to dexamethasone. However, significant differences were observed in positive ion mode. Based on mass spectral analysis, we postulated the unknown peak to be a 6-methyl-16-nor isomer of dexamethasone. Following the procurement of a commercial 9F6MP reference material, the unknown peak was successfully identified as this substance. Interestingly, previous research predicted a high potential for glucocorticoid and anti-inflammatory activity for 9F6MP. However, the therapeutic use of 9F6MP in camels has not been approved by any authorities, and any toxicities and side-effects potentially caused by 9F6MP have not been thoroughly evaluated. Therefore, the misuse of 9F6MP should be strictly controlled for the sake of animal welfare and the integrity of camel racing. The information described in this case report will be beneficial for other anti-doping laboratories in both human and animal sports for the purpose of doping control.
Semisynthetic Cannabinoids Detected in Cannabis-Derived Products in Poland: Statistical Overview, Analytical Challenges, and Legal Interpretative Considerations.
Wrzesień-Tokarczyk W, Masier K, Byrska B … +2 more , Stanaszek R, Sekuła K
Drug Test Anal · 2026 Feb · PMID 41386669 · Publisher ↗
Semisynthetic cannabinoids (SSCs) are a novel group of psychoactive substances obtained by chemical modification of phytocannabinoids such as Δ-THC and CBD. Since 2022, their prevalence has rapidly increased on the Europ...
Semisynthetic cannabinoids (SSCs) are a novel group of psychoactive substances obtained by chemical modification of phytocannabinoids such as Δ-THC and CBD. Since 2022, their prevalence has rapidly increased on the European illicit drug market, including Poland, where they are mainly detected in cannabis-derived products (plant material, resinous products, e-liquids, and edibles). In this study, 1186 cannabis-type samples seized in Poland (2022-2024) were analyzed at the IFR using GC-MS, UHPLC-PDA, and LC-QTOF/MS. SSCs were found in 113 samples (9.5%), mostly in plant material and resinous products. The predominant compound was hexahydrocannabinol (HHC), consistently detected as a mixture of two epimers, with (9R)-HHC prevailing (typical (9R):(9S) ratio 2.0-2.8:1). In most cases, SSCs were applied to plant material with low Δ-THC content, mainly Chemotypes II and III (approximately 60% of all cases), indicating intentional enrichment of material with limited psychoactive potential. Following legislative changes introduced in Poland in 2023, identifications of controlled SSCs decreased; however, the diversity of noncontrolled compounds increased. Co-occurrence of multiple SSCs, their structural similarity, stereoisomerism, and limited availability or delays in obtaining certified reference standards may complicate routine analysis. Each analytical technique presents specific limitations: GC-MS can cause degradation of acetate forms of SSCs; UHPLC-PDA faces challenges in differentiating compounds with similar UV spectra, whereas LC-QTOF/MS cannot fully distinguish structural or stereoisomeric forms. This study highlights the dynamic nature of the SSC market and the importance of advanced, multimethod analytical approaches for reliable identification.
Manhole Sampling at Gyms and Sports Centers in Denmark: A Pilot Study Using Wastewater Analysis as a Tool for Monitoring the Use of Performance-Enhancing Drugs.
Heinsvig PJ, Holler KR, Christiansen AV … +1 more , Lindholst C
Drug Test Anal · 2026 Mar · PMID 41386665 · Publisher ↗
In this study, we assess the feasibility of wastewater sampling from manholes as a potential monitoring tool for performance and image-enhancing drugs (PIEDs) at local gyms and general sports centers in Denmark. Wastewat...
In this study, we assess the feasibility of wastewater sampling from manholes as a potential monitoring tool for performance and image-enhancing drugs (PIEDs) at local gyms and general sports centers in Denmark. Wastewater samples from two gyms and two sports centers were analyzed using two LC-MS-based analytical methods (UPLC-MS/MS and UPLC-TOF-MS) to detect the presence of PIEDs. Samples were collected at 3-h intervals during the opening hours on three selected days of the week for each location. The study showed the presence of anabolic agents such as oxandrolone, stanozolol, and ostarine in wastewater sampled from the gyms. All three compounds are class C drugs in the United States, and likewise, illegal to sell and possess in Denmark. Other PIEDs, such as aromatase inhibitors and psychoactive drugs, were detected in several samples from both gyms and sports centers. However, gyms showed a higher occurrence of PIEDs, whereas general sports centers showed a lower occurrence. Although it cannot be used as a traditional prevalence measure, wastewater sampling from manholes is a useful tool for studying the occurrence of PIEDs among recreational athletes at specific locations or events. Wastewater analysis may be used as an easy and cost-effective tool to complement both preventive and control measures, such as information campaigns, surveys, or personal testing. However, not all locations are suitable for manhole sampling, and it is necessary to consider the legal, ethical, and practical aspects before using wastewater analysis to monitor communities or specific populations.
Further Insights Into the Metabolism of LGD-4033 in Human Urine. Part 1. Structure Elucidation of Additional Important Metabolites.
Angelis YS, Sakellariou P, Thevis M … +3 more , Thomas A, Petrou M, Pitsinos EN
Drug Test Anal · 2026 Feb · PMID 41367083 · Full text
This study presents LC-HRMS/MS analyses of LGD-4033 post-administration urine samples, following hydrolysis with β-glucuronidase and liquid-liquid extraction, against chemically synthesized molecules that matched previou...
This study presents LC-HRMS/MS analyses of LGD-4033 post-administration urine samples, following hydrolysis with β-glucuronidase and liquid-liquid extraction, against chemically synthesized molecules that matched previously proposed metabolites, characterized by H and C NMR. Using this targeted metabolic investigation approach and the direct comparison of retention times and mass spectral data (high-resolution full scan mass accuracy and collision-induced fragmentation patterns), in accordance with WADA's TD2023IDCR provisions, resulted in unambiguous structural elucidation of additional LGD-4033 metabolites, including (a) the epi-long-term dihydroxylated metabolite (M5a); (b) the epi-pyrrolidinone metabolite (M2d); (c) the (R,R)-diastereoisomer of the ring-opened hydroxylated metabolite (M4b); and (d) one of the two detected tris-hydroxylated metabolites (M6a). Additionally, a new, previously undescribed metabolite, which is a hydroxylated derivative of the pyrrolidinone metabolite M2c, was also detected up to 4 days post-administration and coded as M7. Metabolites M5a and M2d are detectable up to 21 days post-administration and can be considered additional long-term markers. These findings expand current knowledge of LGD-4033 metabolism. From a doping control perspective, the proposed synthetic pathways may facilitate the production of reference materials for the detection and identification of a more comprehensive metabolite profile that will increase metabolic certainty in future LGD-4033 adverse analytical findings.
Evaluation of an Innovative Portable Heroin Electrochemical Sensor for Empowering Forensic Laboratories.
Montiel NF, Mazurków J, Van Echelpoel R … +5 more , Daems E, Balcaen M, Deconinck E, Van Durme F, De Wael K
Drug Test Anal · 2026 Feb · PMID 41367054 · Publisher ↗
The increasing misuse of opioids in Europe is an alarming trend, leading to severe social and health consequences. Heroin, a highly potent and addictive opioid, remains the main contributor to the health burden associate...
The increasing misuse of opioids in Europe is an alarming trend, leading to severe social and health consequences. Heroin, a highly potent and addictive opioid, remains the main contributor to the health burden associated with opioid use in the region. Illicit drug characterization and profiling offer valuable insights into the complexity of heroin seizures, assisting law enforcement agencies and forensic experts in gathering evidence for legal proceedings. This study provides a comprehensive overview of the composition of heroin seizures and assesses the feasibility of an electrochemical fingerprint approach for the detection of heroin and its associated components. In the initial phase, the primary focus was on developing an electrochemical sensor optimized for heroin detection. The sensor's performance was validated using street samples provided by Sciensano, a Belgian health institute, ensuring its accuracy and reliability in identifying heroin. Once the capabilities of the sensor were demonstrated, the discrimination of alkaloids and cutting agents in seized samples was integrated into a customized software script. Subsequently, an extensive validation process was conducted using a new dataset of heroin seizures from the Belgian National Institute for Criminalistics and Criminology. The follow-up verification confirmed the sensor's effectiveness in detecting heroin, cutting agents, and alkaloids, highlighting its potential as a valuable tool for drug profiling. This portable, user-friendly device with automatic readout could become essential for forensic experts, law enforcement, and harm reduction efforts in addressing the opioid crisis.
Assessing the Effect of Probiotics in the Steroidal Module of the Athlete's Biological Passport.
Merlo ABM, Piper T, Lobigs L … +4 more , de Figueiredo M, Rhumorbarbe D, Thevis M, Robinson N
Drug Test Anal · 2026 Feb · PMID 41340503 · Publisher ↗
Athletes are increasingly using probiotic supplementation to support their overall health, and it can be particularly beneficial for female athletes in managing recurrent urinary tract infections and bacterial vaginosis....
Athletes are increasingly using probiotic supplementation to support their overall health, and it can be particularly beneficial for female athletes in managing recurrent urinary tract infections and bacterial vaginosis. One route of probiotic administration for females is vaginal application, which enables direct modulation of the microbiota. While probiotics are widely recognised for their health benefits, their potential impact on urinary steroidal markers monitored in the Steroidal Module of the Athlete Biological Passport remains unexplored. Given the biological overlap between vaginal and urinary microbiomes, bacteria from vaginal probiotics could transfer into urine samples, potentially altering steroid profiles through microbial enzymatic activity. This study investigates whether vaginal probiotic use, specifically Lactobacillus reuteri and plantaraium, could influence urinary steroid markers relevant to the steroidal passport. In vitro and in vivo approaches were employed to evaluate the potential effects of contamination and variability on key steroidal markers. Analyses of in vitro and in vivo experiments suggest that vaginal probiotics do not substantially affect urinary steroid markers monitored in the Athlete Biological Passport. However, some variations were observed that merit further investigation. These findings contribute to a better understanding of how vaginal probiotics might interfere with doping control results, emphasising the need for further research to ensure accurate interpretation of urine steroidal profiles in the female athlete.
Simulation of the Metabolism of New Psychoactive Substances Using Electrochemistry-Mass Spectrometry: Introducing an Innovative Software Tool for Rapid Data Evaluation.
Wesner M, Heuckeroth S, Pütz M … +1 more , Karst U
Drug Test Anal · 2026 Feb · PMID 41308614 · Full text
An innovative software tool for the rapid and efficient simulation of the metabolism of new psychoactive substances (NPS) was developed, based on the open-source project mzmine, and applied. NPS are compounds designed to...
An innovative software tool for the rapid and efficient simulation of the metabolism of new psychoactive substances (NPS) was developed, based on the open-source project mzmine, and applied. NPS are compounds designed to mimic the psychotropic effects of established illicit drugs while circumventing drug legislation. These compounds are developed solely regarding their desired effects, thus possibly leading to harmful side effects including the formation of toxic metabolites. Analytical reference standards, needed to carry out metabolic studies, are not immediately available because emerging NPS are primarily discovered subsequent to drug confiscations. Using these confiscated substances in traditional metabolic in vivo or in vitro studies is often not possible due to the substances being impure or being a part of a mixture of different NPS. Therefore, a software tool was developed to streamline the evaluation of data acquired by the online combination of electrochemistry and mass spectrometry for the simulation of NPS metabolism. Using this tool, it is possible to generate mass voltammograms directly from mass spectrometric raw data. Combining this newly implemented tool with existing filtering algorithms in mzmine, we simulated the metabolism of the synthetic cannabinoid receptor agonist (SCRA) methyl 3,3-dimethyl-2-[1-(pent-4-en-1-yl)-1H-indazole-3-carboxamido] butanoate (MDMB-4en-PINACA) from a mixed solution of different NPS. Fragmentation data indicated that one of the transformation products found for MDMB-4en-PINACA is likely of a quinoid structure. The potential formation of this possibly highly reactive quinoid metabolite could be a first hint for possible causes of adverse side effects frequently reported after the recreational use of MDMB-4en-PINACA and related SCRAs.
Improvement of EPO Transgene Detection From Polymeric Dried Blood Spots for Antidoping Application.
Marchand A, Roulland I, Ericsson M
Drug Test Anal · 2026 Feb · PMID 41308298 · Full text
For the past couple of years, black market products have appeared and were confirmed to contain genetic products coding for human erythropoietin (EPO). While being prohibited by the World Anti-Doping Agency (WADA), they...
For the past couple of years, black market products have appeared and were confirmed to contain genetic products coding for human erythropoietin (EPO). While being prohibited by the World Anti-Doping Agency (WADA), they could be used to produce endogenously more EPO hormone and hence increase performance. In a previous work, we demonstrated the potential of 20-μL dried blood spots (DBS) to detect the presence of EPO transgene in human blood down to 250 copies (12,500 copies/mL), despite lower sensitivity (30-fold) than in 1-mL fresh blood. As the use of DBS as a collection matrix for antidoping is going to expand in the near future, our aim was to develop and validate a new protocol to improve the sensitivity of gene doping detection from DBS. Three DBS devices were evaluated: polymeric Tasso-M20 (TASSO Inc.) and Mitra (Neoteryx), and cellulosic Protein Saver 903 (Whatman). The best results were achieved with polymeric DBS, and a full validation was performed for the detection of the EPO transgene using Tasso M-20 DBS; 1500 copies/mL were detected in 50% of cases and robust detection was obtained at 5000 copies/mL (100 copies transgene in 20-μL DBS) with the four spots of the Tasso device tested over several weeks. The results confirm that polymeric DBS can be used as an alternative to fresh blood for gene doping detection with high sensitivity simplifying also potential reanalysis in the future.
Rapid Identification of New Psychoactive Substances in Letters by LA-APCI-MS.
Wesner M, Rämisch H, Besch L … +2 more , Schmeinck J, Karst U
Drug Test Anal · 2026 Feb · PMID 41285127 · Full text
New psychoactive substances (NPS), especially synthetic cannabinoid receptor agonists (SCRA), are increasingly smuggled into prisons via infused mail. Consumption of those substances by inmates in prisons is associated w...
New psychoactive substances (NPS), especially synthetic cannabinoid receptor agonists (SCRA), are increasingly smuggled into prisons via infused mail. Consumption of those substances by inmates in prisons is associated with increased aggression, violence, and organized crime. Onsite detection of infused mail often is challenging. Because the infused papers do not show any visible stains or olfactory changes, physical inspection is often insufficient. The applicability of further conventional on-site detection methods like immunoassays and sniffer dogs is severely limited. Because of the rapidly changing supply of already circulating and newly emerging NPS, it is impractical to impossible to keep up with the development of immunoassays or the training of sniffer dogs. Hence, confiscated mail samples are routinely tested by either liquid chromatography or gas chromatography coupled to mass spectrometry (MS), which is costly and time-consuming. In this study, recent advancements in the hyphenation of laser ablation (LA) and molecular MS were investigated regarding the possible application for the rapid and easy detection of NPS in prison mail. Utilizing an in-house developed LA-MS hyphenation based on atmospheric pressure chemical ionization (APCI), 31 mail samples confiscated in German prisons were analyzed. It was possible to correctly identify 27 samples containing SCRAs. For these positive samples, it was also possible to detect the specific compounds each paper was infused with. The use of LA-APCI-MS has simplified sample preparation and reduced analysis time per sample to 1 min.
Characterization of Novel Enarodustat Metabolites Using Liquid Chromatography-High Resolution Mass Spectrometry for Doping Control Purposes.
Hou J, Zhang L, Wang Z … +1 more , Yang S
Drug Test Anal · 2026 Feb · PMID 41276953 · Publisher ↗
Hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs) represent a novel class of therapeutic substances that increase erythropoiesis. Due to their performance-enhancing effects and potential risk of abuse, th...
Hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs) represent a novel class of therapeutic substances that increase erythropoiesis. Due to their performance-enhancing effects and potential risk of abuse, these agents were added to the World Anti-Doping Agency (WADA) Prohibited List in 2011. Enarodustat is a novel HIF-PHI and has been approved for clinical use in China in 2023. This study primarily aimed to characterize its major urinary metabolites for antidoping purposes. A single oral dose of 40-mg enarodustat was administered to a volunteer. Urine samples were collected over 28 days and processed using solid-phase extraction (SPE). Analytical methods included liquid chromatography-high resolution mass spectrometry (LC-HRMS) under both positive and negative electrospray ionization conditions, complemented by in vitro metabolism studies using human liver microsomes (HLMs) for the characterization and identification of metabolites. A total of eight metabolites were detected, including Phase I products such as parent compound (PC) isomer (M1), monohydroxylation (M2), dihydroxylation (M3), and dehydrogenation (M4) metabolites, as well as Phase II conjugates involving methylation (M5), glycosylation (M6), glucuronidation (M7), and monohydroxylation-sulfation (M8) in vivo. Among these, M3-M5 are novel metabolites. In addition, compared with other metabolites, PC and M2 exhibited longer detection windows, suggesting they are valuable biomarkers for doping control purposes. The study elucidates enarodustat's metabolic pathways and provides a foundation for developing sensitive detection methods. Future work should focus on synthesizing reference materials to identify metabolite structures.
Surveillance of Phenibut in Wastewater During a Brazilian Carnival.
Gomes BRS, de Oliveira AFB, Vieira AM … +3 more , Nadarajan D, Bade R, Santos JM
Drug Test Anal · 2026 Feb · PMID 41276941 · Full text
Phenibut is a new psychoactive substance (NPS) first synthesized in Russia in 1963 as a derivative of gamma-aminobutyric acid. Originally developed for therapeutic use, it has gained popularity for nonmedical purposes, i...
Phenibut is a new psychoactive substance (NPS) first synthesized in Russia in 1963 as a derivative of gamma-aminobutyric acid. Originally developed for therapeutic use, it has gained popularity for nonmedical purposes, including recreational and cognitive enhancement. In Brazil, phenibut is uncontrolled and easily purchased online. This study used wastewater-based epidemiology (WBE) to investigate phenibut use patterns in two northeastern Brazilian cities. Composite daily wastewater samples were collected from two treatment plants (WWTPs), Recife (WWTP) and Olinda (WWTP), during two periods in 2023: Carnival and a reference week. Samples underwent solid-phase extraction (SPE) and analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Phenibut concentrations were converted to population-normalized mass loads (PNMLs, mg/day/1000 inhabitants). The highest phenibut levels and PNMLs (up to 4.06 mg/day/1000 inhabitants) occurred during Carnival at WWTP, located in a major tourist area, suggesting recreational use. During the reference week, PNMLs ranged from detection limits to 2.29 mg/day/1000 inhabitants on weekdays, indicating possible functional or cognitive enhancement use. These findings reveal two distinct use patterns: recreational peaks during Carnival weekends and possible functional use on weekdays outside festive periods. This is the first evidence of phenibut detection in Brazilian wastewater and its temporal use patterns. The results highlight WBE's value in monitoring NPS trends and suggest recreational use predominates during large events. This underscores the need for public health attention and regulatory monitoring of uncontrolled substances with abuse potential.
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