{"title":"Identification of Three Novel Tetrahydrocannabinol Analogs in the European Market.","authors":"Evangelos Dadiotis, Sotiris Mpakaoukas, Vangelis Mitsis, Eleni Melliou, Prokopios Magiatis","doi":"10.1002/dta.3866","DOIUrl":"https://doi.org/10.1002/dta.3866","url":null,"abstract":"<p><p>Synthetic cannabinoids, known as Spice or K2, emerged in Europe and the United States between 2005 and 2008, peaking in incidents by 2015 with severe health implications. In 2021, the identification of hexahydrocannabinol (HHC), a semisynthetic cannabinoid (SSC), led to regulatory control in more than 20 countries in Europe, several US states, and other jurisdictions. A 2024 study in the United States highlighted the diversity of semisynthetic cannabinoids in the US market. New entries of SSCs are increasingly available in the European market, often found as blends in consumer products. This highlights the growing complexity of their regulation and the potential public health risks due to limited toxicological data. The major ingredients, isolated from \"CB9\", \"tresconol\", and \"CBx\", were subjected to mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy. The major isolated ingredient of \"CB9\" was identified as [2-(E)-propen-1-yl]-Δ<sup>8</sup>-tetrahydrocannabinol-acetate. The major ingredient of \"tresconol\" was identified as [2-propen-2-yl]-Δ<sup>9</sup>-tetrahydrocannabinol, and the major ingredient of \"CBx\" was identified as [2-propen-2-yl]-Δ<sup>8</sup>-tetrahydrocannabinol. These compounds have no available spectroscopic or chromatographic data, have never been identified in Cannabis plants, and cannot be identified by standard chromatographic forensic analytical methods, without the use of spectroscopic techniques due to the lack of reference standards. The products were complex mixtures of previously unknown synthetic cannabinoids lacking established safety profiles. These findings highlight the potential public health risks associated with unregulated SSCs, similar to the concerns raised during the 2015 Spice outbreak. The presence of these novel substances requires careful monitoring to prevent future health crises.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143254153","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":"Impact of Menstrual Cycle and Oral Contraceptives on Haematological and Inflammatory Biomarkers in Highly Trained Female Athletes.","authors":"Katia Collomp, Caroline Teulier, Carole Castanier, Juliette Bonnigal, Alexandre Marchand, Corinne Buisson, Magnus Ericsson, Nathalie Crépin, Emmanuelle Duron, Eric Favory, Mathieu Zimmermann, Virgile Amiot, Agnès Olivier","doi":"10.1002/dta.3859","DOIUrl":"https://doi.org/10.1002/dta.3859","url":null,"abstract":"<p><p>Haematological and inflammatory biomarkers play an important role in athlete performance and health, with some of them used in the fight against doping. However, little is known about how they are modulated by sex hormone fluctuations in highly trained female athletes. We therefore measured the haematological parameters monitored in the athlete biological passport (ABP) as well as erythropoietin, serum markers of iron and inflammatory statuses (iron, ferritin, transferrin, transferrin saturation, albumin, creatinine, total protein, interleukin-6 and TNF-alpha) in 20 highly trained female athletes: 10 with normal menstrual cycle (NMC) during the early follicular and mid-luteal phases and 10 using a combined oral contraceptive (COC, i.e., ethinyloestradiol and levonorgestrel) during active and inactive hormone intake. Body composition, leptin and lipid profile (total cholesterol, HDL, LDL and triglycerides) were determined in parallel. No changes were observed throughout NMC phases. Irrespective of active/inactive pill intake, COC use increased transferrin, triglycerides as well as reticulocyte count (p < 0.05) and decreased interleukin-6 (p < 0.05), with no significant changes in the other parameters studied. In conclusion, given our results across NMC phases in highly trained athletes, it seems warranted to investigate whether intense physical training would mitigate the impact of endogenous sex hormones on body composition and haematological and inflammatory parameters. In addition, further studies are needed to determine the extent of the changes induced by COCs on these blood biomarkers in elite female athletes when subjected to extreme environments such as intensive training or competition in humid heat, cold and/or hypoxia or when using other medications in parallel.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143254190","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":"LC-IRMS: Towards a High-Throughput IRMS Methodology?","authors":"Michaël Polet, Lenka Honesova, Peter Van Eenoo","doi":"10.1002/dta.3858","DOIUrl":"https://doi.org/10.1002/dta.3858","url":null,"abstract":"<p><p>Recently, an LC-IRMS methodology has become available for analyzing endogenous anabolic steroids in a urinary matrix. This could potentially open up new possibilities such as for example a high throughput IRMS methodology.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143254212","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}
Rafael D Soares, Danielle K John, Marcos P Thomé, Patrícia S Corrêa, Klester S Souza, Marco F Ferrão
{"title":"Robust Detection of Ecstasy-Like and Adulterants Through ASAP-MS and DD-SIMCA.","authors":"Rafael D Soares, Danielle K John, Marcos P Thomé, Patrícia S Corrêa, Klester S Souza, Marco F Ferrão","doi":"10.1002/dta.3860","DOIUrl":"https://doi.org/10.1002/dta.3860","url":null,"abstract":"<p><p>Ecstasy is a complex and hazardous substance, and its identification is increasingly challenging. Conventional analytical methods have limitations in terms of sensitivity and selectivity, and more precise techniques are time-consuming and necessitate sample preparation. ASAP-MS and DD-SIMCA are two methods that have the potential to address these issues. This research delves into the efficacy of ASAP-MS and DD-SIMCA as a rapid and dependable approach for detecting ecstasy and its adulterants. PCA was conducted as an initial exploration, with the first three principal components (PCs) capturing 69% of the overall data variability. The score plot of PC1 × PC3 revealed the distribution of samples containing MDA and MDMA. The DD-SIMCA model exhibited high sensitivity in identifying the target samples (MDA) and relatively high specificity in training and test sets. These results underscore the effectiveness of ASAP-MS, PCA, and DD-SIMCA for precise identification of ecstasy and its adulterants, indicating their potential in drug identification and analysis. We observed that the chemometric model associated with ASAP-MS was able to accurately identify, when compared to the results obtained by the standard technique, the constituents of ecstasy tablets, even in the presence of adulterants. Furthermore, the method could detect emerging psychoactive substances that are typically not targeted by traditional analytical approaches. These findings suggest that ASAP-MS and DD-SIMCA could be valuable tools in forensic drug analysis laboratories. The method is rapid, reliable, and versatile for identifying a wide range of ecstasy and its adulterants.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143187802","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":"Administration Studies in Equine Antidoping Research: Designing Scientific Investigations to Effectively Direct Medication Control in Racehorses.","authors":"Heather K Knych","doi":"10.1002/dta.3857","DOIUrl":"https://doi.org/10.1002/dta.3857","url":null,"abstract":"<p><p>Pharmacokinetics is the study of the movement of drug in the body and includes the processes of absorption, distribution, metabolism, and excretion. Pharmacodynamics is the pharmacologic effect of the drug on the body. The pharmacokinetics of a drug determines its pharmacologic effect. Pharmacokinetic studies describe drug concentrations while pharmacodynamics allow for assessment of drug effects. Combined pharmacokinetic/pharmacodynamic studies allow for integration of drug concentrations with pharmacologic effect. Data generated from pharmacokinetic studies can be especially useful in establishing regulatory recommendations, determining appropriate thresholds, screening limits, administrative stand down times, and corresponding detection times. To generate the appropriate information, the following must be considered (1) the test subjects (i.e., number, age, breed, and fitness level), (2) selection of an appropriate dose/route and duration of administration, (3) sample matrix (blood, urine, and hair), (4) time(s) of sample collection, (5) development of an analytical method with appropriate sensitivity, and (6) what analytes to measure (parent and/or metabolite). Pharmacokinetic studies generate drug concentration data that can be used to calculate key pharmacokinetic variables necessary for establishing screening levels and detection times. Pharmacodynamic assessments can aid in understanding the pharmacologic effects of drugs and in correlating drug concentrations to these effects. Various models, including in vivo (whole animal), in vitro, and ex vivo assessments, can be utilized to determine pharmacodynamic effects. Factors to consider in the design of pharmacokinetic studies, basic pharmacokinetic parameters, and examples of pharmacodynamic assessments will be discussed in detail during this tutorial.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143057438","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":"Cost Minimized Immunoaffinity Purification of EPO and Its Analogs in Doping Control-A Step-by-Step Protocol for Human Urine and Blood.","authors":"Christian Reichel, Günter Gmeiner, Mario Thevis","doi":"10.1002/dta.3848","DOIUrl":"https://doi.org/10.1002/dta.3848","url":null,"abstract":"<p><p>A cost minimized immunoaffinity protocol was developed, which allows the direct purification of ERAs (urinary and recombinant human EPO, Darbepoetin, EPO-Fc, CERA) from human urine. The method applies magnetic beads and needs no covalent immobilization of the capture antibody. It requires only 10 mL of urine, 1 μg of anti-EPO antibody, and 25 μL of bead slurry. The beads are coated with the capture antibody in advance and can be stored in the refrigerator for months without any loss of functionality. The protocol was fully validated in combination with SAR-PAGE and Western blotting using the biotinylated clone AE7A5 anti-EPO antibody. It is compliant with the criteria of TD2024EPO of the World Anti-Doping Agency (WADA) for the gel electrophoretic detection of ERAs. For each ERA, the achieved limit of detection (LOD) is at least one tenth of the minimum required performance limit (MRPL) of WADA, that is, 0.1 IU/L, 0.1 pg/mL, 0.5 pg/mL, and 0.5 pg/mL for rEPO, Darbepoetin, EPO-Fc, and CERA, respectively. After slight modification, the protocol is also applicable to serum and plasma and also fulfils the corresponding MRPLs of WADA for these matrices. Compared to commercial immunoaffinity purification kits for EPO, the material costs are significantly lower but with identical results.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143051150","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}
Sergi Coll, Deamelys Hernández, Claudia Bressan, Rosalia Ramírez, Núria Monfort, Ana Aldea-Perona, Rosa Ventura
{"title":"Impact of Repeated Glucocorticoid Oral Administration on the Urinary Steroid Profile.","authors":"Sergi Coll, Deamelys Hernández, Claudia Bressan, Rosalia Ramírez, Núria Monfort, Ana Aldea-Perona, Rosa Ventura","doi":"10.1002/dta.3853","DOIUrl":"https://doi.org/10.1002/dta.3853","url":null,"abstract":"<p><p>The detection of endogenous anabolic androgenic steroids (EAAS) is performed with the Steroidal Module of the Athlete Biological Passport (ABP). Glucocorticoids (GC) could be a confounding factor to the ABP Steroidal Module because they inhibit the hypothalamic-pituitary-adrenal axis, and ABP metabolites have partial adrenal origin. In previous studies, single-dose systemic GC administrations have been shown to reduce the urinary ratios A/T and 5αdiol/E. In this work, the impact of repeated oral doses of GCs on the urinary steroid profile (SP) has been evaluated. The treatments administered consisted of multiple oral administrations of methylprednisolone (12 mg/24 h for 3 days, n = 8) and dexamethasone (2 mg/12 h for 5 days, n = 8). Urine samples were collected before, during, and after the GC treatments, and the SP was measured in all samples using gas chromatography-tandem mass spectrometry. The multiple-dose oral administration of GCs resulted in a treatment-dependent reduction of the excretion rates of some urinary metabolites (5αAdiol, A, and Etio) and the urinary SP ratios A/T and 5αAdiol/E. The T/E ratio was not significantly affected. Overall, although the consumption of GC could result in atypical profiles for A/T and 5αAdiol/E ratios, according to the cost/benefit assessment, GC should not be considered a confounding factor to the urinary SP because misunderstandings would only take place in very specific situations and, in those cases, the analysis by isotope ratio mass spectrometry of the urine triggering the atypical profile would demonstrate the endogenous origin of the SP metabolites.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143031678","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":"Rapid and Simple Dispersive Liquid-Liquid Microextraction (DLLME) Sample Preparation for Propofol Analysis in Hair, Blood, and Urine by Gas Chromatography-Mass Spectrometry.","authors":"Sara Odoardi, Serena Mestria, Valeria Valentini, Giulia Biosa, Sabina Strano Rossi","doi":"10.1002/dta.3856","DOIUrl":"https://doi.org/10.1002/dta.3856","url":null,"abstract":"<p><p>Propofol is a widely used anesthetic. Although considered safe, propofol-related deaths occur, as it is sometimes abused recreationally or used to commit suicide. A simple, rapid, and reliable method for its analysis in various biological samples is needed. Sample clean-up is a critical step in the analysis, both in terms of time and cost, indeed. Dispersive liquid-liquid microextraction (DLLME) is a simple and fast extraction based on ternary solvent mixtures that uses small volumes of solvent and sample. A DLLME extraction followed by gas chromatography-mass spectrometry (GC-MS) analysis was developed and validated for the analysis of propofol in blood, urine, and hair. The same extraction mixture of 2.5:1 methanol/chloroform was used for the different biological samples. Validation for linearity, LOD, LOQ, precision, accuracy, and recovery gave satisfactory results for the three types of biological samples included in the study, with limits of quantification of 1 μg/mL for urine, 0.2 μg/mL for blood, and 0.1 ng/mg for hair. The DLLME procedure for purification involves a small amount of solvent, thus reducing the cost and the environmental impact. In addition, a high enrichment factor is obtained, and the time for analysis is short. The method was applied to authentic post-mortem samples for the determination of propofol in blood, urine, and hair. Also, segmental hair analysis was performed to assess chronic propofol abuse. The developed method proved to be rapid, simple, and cost-effective for blood, urine, and hair extract clean-up for the determination of propofol by GC/MS.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143021345","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}
Marco Ballotari, Nicola Pigaiani, Anna Bacci, Karen S Scott, Gregory G Davis, Rossella Gottardo, Federica Bortolotti
{"title":"Retrospective Evaluation of Novel Synthetic Opioids and Xylazine Chronic Intake by Post-Mortem Hair Testing.","authors":"Marco Ballotari, Nicola Pigaiani, Anna Bacci, Karen S Scott, Gregory G Davis, Rossella Gottardo, Federica Bortolotti","doi":"10.1002/dta.3852","DOIUrl":"https://doi.org/10.1002/dta.3852","url":null,"abstract":"<p><p>Fentanyl and its derivatives (nonpharmaceutical fentanyl, NPFs) represent the largest group among synthetic opioids. Fentanyl-related deaths and fatalities from tampering with pharmaceutical products have been reported. Furthermore, in the United States, adulterants such as xylazine and other substances, including the nitazenes class of opioids, have been found in an increasing number of unintentional overdose deaths, drug seizures, and reports of use by recreational drug users. Monitoring the diffusion of fentanyl, NPFs, nitazenes, and adulterants among the population is a fundamental pursuit in forensic toxicology. The use of hair analysis is perfect for this purpose, providing essential information regarding previous intake or exposure to xenobiotics. The present study focused on the development and validation of a UPLC-MS/MS method for the detection and quantification of fentanyl, NPFs, and xylazine, as well as the semiquantitative detection of nitazenes in hair samples from post-mortem cases collected under the jurisdiction of the Jefferson County Coroner/Medical Examiner's Office (Birmingham, AL, USA). The method was validated according to international guidelines and applied to the analysis of n = 250 post-mortem hair samples. In 52% of the analyzed hair samples, fentanyl, its main metabolites, and related analogs were detected, showing significant exposure to these substances in the population. Moreover, xylazine was detected in n = 48 hair samples (19.2%). The developed UPLC-MS/MS method proved suitable for rapid chromatographic separation and sensitive detection of the studied compounds. In addition, this is the first time that xylazine and protonitazene have been measured in hair samples of subjects exposed to synthetic opioids.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142996926","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}
Selina Hemmer, Maximilian Ninnig, Lea Wagmann, Sascha K Manier, Markus R Meyer
{"title":"Comprehensive Qualitative Drug Screening in Emergency Toxicology Using an Automated LC-MS<sup>n</sup> System:Simultaneous Quantification of Relevant Drugs and Metabolites in Blood Plasma.","authors":"Selina Hemmer, Maximilian Ninnig, Lea Wagmann, Sascha K Manier, Markus R Meyer","doi":"10.1002/dta.3855","DOIUrl":"https://doi.org/10.1002/dta.3855","url":null,"abstract":"<p><p>Rapid and comprehensive qualitative and quantitative analytical procedures are crucial in 24/7 emergency toxicology (ET) to support diagnosis and treatment of acute intoxications and to monitor their progression and efficacy of detoxification strategies. This study aimed to develop the simultaneous qualitative and quantitative analysis of 62 drugs, as well as seven active metabolites in human blood plasma using an automated liquid chromatography (LC)-linear ion trap mass spectrometry (MS) screening system. Sample preparation was conducted by liquid-liquid extraction, and plasma concentrations were determined using an electronically stored 5-point calibration. Validation was performed according to international guidelines and recommendations for ET including selectivity, carry-over, accuracy, precision, and matrix effects. Finally, applicability was evaluated using case samples and proficiency tests. The method demonstrated selectivity for all analytes, with no significant carry-over or matrix effects. Accuracy and precision recommended for ET could be fulfilled for all tested analytes, except for 10 analytes. Patient plasma samples were analyzed and compared with results obtained by reference LC-tandem MS or gas chromatography-MS methods. Furthermore, the applicability of the method could be demonstrated. It provides a fast, robust, and reliable blood plasma screening for 69 analytes and an additional quantification of 59 analytes relevant in ET. The use of an electronically stored 5-point calibration and a simplified \"push and print solution\" allows for straightforward assessment of blood plasma levels of the analytes.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2025-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142996917","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}