Drug Testing and Analysis最新文献

With the discovery of the variant c.577del in the EPO gene, the procedure for detecting the presence of recombinant erythropoietin (rEPO) has become complicated and time-consuming. To address this situation, an rEPO confirmation method that uses reverse-normal immunopurification coupled with western blotting (WB) and sodium N-lauroylsarcosinate polyacrylamide gel electrophoresis (SAR-PAGE), which can detect the EPO variant (VAR-EPO) and rEPO with anti-VAR-EPO and anti-EPO antibodies, has been developed. Therefore, it is necessary to develop an internal standard (IS) that can be recognized by an anti-VAR-EPO antibody to monitor reverse immunopurification, ensuring the reliability and accuracy of the analysis. In this study, we constructed an IS based on VAR-EPO modified with polyethylene glycol (PEG) and then assessed its reliability and applicability in doping analysis. The data provided here show that the obtained PEGylated VAR-EPO can be used as an IS to monitor the detection of not only rEPO and VAR-EPO but also EPO receptor agonists in urine samples.

The classical psychedelic drug (+)-lysergic acid diethylamide (LSD) continues to attract considerable multidisciplinary interest, and over the last eight decades, many derivatives and analogs of LSD have been synthesized. One site on the ergoline scaffold of LSD that has been frequently modified is the N¹-position, with the N¹-acylated LSD derivative 1-acetyl-LSD (1A-LSD, ALD-52) being one of the earliest examples. In more recent years, several other alkylcarbonyl- and cycloalkylcarbonyl-substituted LSD derivatives have been evaluated, including several distributed as research chemicals. Although N¹-substitution is detrimental for the activity of LSD at the 5-HT_2A receptor (the primary site of action of psychedelic drugs), N¹-acylated LSD derivatives are rapidly hydrolyzed in vivo and are believed to act as prodrugs for LSD. Recently, 1-(thiophene-2-carbonyl)-LSD (1T-LSD, SYN-L-021) was detected as a new recreational drug, signaling a move towards N¹-acyl groups with an aromatic character. The present study was conducted to investigate the analytical profile and pharmacology of 1-(2-furoyl)-lysergic acid diethylamide (1F-LSD, SYN-L-005), a novel analog of 1T-LSD. The binding of 1F-LSD to the 5-HT_2A receptor and other monoamine sites was assessed using radioligand binding. Furthermore, the in vivo activities of 1F-LSD and 1T-LSD were assessed in C57BL/6 J mice by comparing their biotransformation to LSD and effects on the head-twitch response (HTR), a 5-HT_2A-mediated behavior. Both 1F-LSD and 1T-LSD induced the HTR in mice and were hydrolyzed to LSD after in vivo administration, indicating that both substances exhibit LSD-like properties and may serve as prodrugs for LSD.

The contamination of materials in urban areas by chemical weapons is a critical issue, especially as these materials can serve as key evidence in forensic investigations. Concrete, commonly found in urban environments, is highly porous and can retain chemical residues. However, its alkaline nature accelerates the degradation of chemical warfare agents, complicating the recovery of usable evidence. This study explores the recovery and persistence of alcohols and thiols, final degradation products of nerve and blistering agents, from two types of concrete matrices: lightweight concrete formworks and dense, steel-reinforced concrete blocks. Using an optimized method, uncrushed concrete fragments (up to 85 g) were extracted with acetone, monitoring two critical parameters: apparent recovery and persistence. The influence of external conditions, such as water addition, temperatures between 5°C and 35°C, and varying airflow speeds (1.7-5.1 m·s^-1), was systematically evaluated. Reference conditions involved dried concrete at 22°C with no airflow. The findings revealed that alcohol recovery aligned with the volatility of the compounds, with denser concrete exhibiting lower recoveries but greater persistence. Thiols quickly converted to disulfides. Notably, temperature and moisture had the most profound effects on the recovery and persistence of the chemicals. These results highlight the importance of considering environmental factors when assessing chemical warfare agents and their degradation products in concrete, offering insights relevant to forensic science, environmental safety, and military defense. The study demonstrates how concrete's properties and external conditions can alter the forensic traceability of chemical contaminants.

The use of alkalinising agents prior to racing for manipulating performance in the horse has been identified since the 1990s. To mitigate the risk, an international threshold for available carbon dioxide in equine plasma based on analyses using the Beckman Synchron EL-ISE analyser was adopted in 1994 by the International Federation of Horseracing Authorities (IFHA) and revised from 37 to 36 mM in 2004. In 2009, the technical support for the above instrument was discontinued by its manufacturer. Based on the same measurement principle (i.e., ion selective electrode), the Beckman DxC600 analyser was selected as an alternative and validated against the protocol developed by the Association of Official Racing Chemists (AORC). Recently, the DxC600 analyser is also no longer supported by Beckman. Various alternative methods for measuring total carbon dioxide (TCO₂) in plasma have been explored. Among these, a headspace gas chromatography-mass spectrometry (HS-GC/MS) method was first reported by the Analytical Forensic Testing Laboratory (AFTL) in 2017. Methods based on the same measurement principle were later developed by different horseracing laboratories. With the objective of cross-validating the new HS-GC/MS methods and to establish an absolute (rather than instrument-dependent or empirical) threshold, an international research collaboration was initiated among different racing laboratories. This paper describes the results of cross-validation studies conducted in November 2019 and December 2022 using horse administration samples from Canada and France, respectively, the determination of a threshold based on population data, and some technical insights on the HS-GC/MS methods.

Anti-doping efforts aim to reduce the prevalence of doping through a combination of education, deterrence, and detection. Detection of doping practices, for example through testing and/or investigations, aims both to catch committed dopers and deter potential dopers. To date, little empirical evidence is available examining the ability of detection strategies to deter athletes from doping. Here, trends in adverse analytical findings (AAFs) for EPO or other EPO-Receptor Agonists (ERAs) were examined over an 8-year period in order to assess the impact of ERA testing and detection on athlete behavior. It was observed that the majority (62.8%) of ERA AAFs occur on samples collected on the day of a competition. Evidence is also presented that the largest fraction of ERA AAFs occurs on the first sample ever taken from an athlete (43.2%), and that the ERA AAF rates decline steadily as athletes continue to be tested. These findings provide evidence of a deterrent effect of testing on ERA use in sport.

Hormonal growth promotants (HGPs) are a class of pharmaceutical agents commonly administered to cattle in the United States to improve growth rates of the animal, alter behavior, or to improve the desired characteristics of retail cuts of meat. There is a concern that low residual concentrations of HGPs may remain in tissue after slaughter, and consumption of tissues containing these compounds may increase the risk of adverse health outcomes, including cancer. Sensitive and selective methods are necessary to assess exposure of HGPs by populations that consume meat products from animals that may have been administered HGPs. A liquid chromatography-tandem mass spectrometry method was developed and validated to detect the low-level presence of HGPs including estradiol, testosterone, estradiol benzoate, melengestrol, melengestrol acetate, progesterone, testosterone propionate, trenbolone, trenbolone acetate, and α-zearalanol in retail cuts of meat following a liquid-liquid extraction using a high pH solution with 30-50× less mass of meat required as compared to similar approaches. Good chromatographic performance and sensitivity was achieved utilizing ammonium fluoride as a mobile phase additive without the need for derivatization. Validation parameters including accuracy, precision, recovery, matrix effects, limits of detection, limits of quantitation, linear range, and stability were determined. The limits of detection ranged from 0.1 to 1.0 ng/g, depending on the compound, with adequate accuracy and precision without the need for extensive sample preparation approaches.

Erythropoietin receptor agonist (ERA) testing for antidoping can be achieved in both urine and blood samples. Recent published work showed the comparability between the two matrices and focused on detectability in microvolumetric capillary serum samples collected using the Tasso+ device. Currently, in the antidoping field, blood samples are required to be shipped under cold, temperature-controlled conditions. However, due to the suggested greater stability of EPO in blood compared to urine, it is believed that blood samples should be viable for ERA analysis if shipped under the ambient, not cold temperature-controlled conditions to which urine samples are subjected. In this collaborative study with the Ultimate Fighting Championship, microvolumetric capillary serum samples were collected in the field and shipped under ambient conditions to the laboratory for ERA analysis. Resulting data showed that endogenous EPO was detectable in 100% of these samples, showing no loss in detectability despite shipping under non-controlled conditions. Further, ERA analyses were conducted in the laboratory on additional in-house collected samples and post-EPO administration samples subjected to various storage and shipping conditions, also showing reliable endogenous and recombinant EPO detectability in all samples except those experiencing extreme temperature (50°C) conditions. Taken together, these data highlight the stability of EPO in blood samples and show that ERA blood samples can be collected in the field and shipped without costly temperature-controlled shipping methods and without a loss in detectability.

The phosphodiesterase 4 (PDE4) inhibitors constitute a relatively modern class of medications that are known for inducing bronchodilation and exhibiting anti-inflammatory properties within the body. Due to these properties, there is concern regarding their potential misuse as performance-enhancing substances in competitive sports. This study delves into the metabolic conversion of roflumilast in thoroughbred horses following oral administration and in vitro experimentation using equine liver microsomes and Cunninghamella elegans. High-performance liquid chromatography coupled with a Q Exactive Orbitrap mass spectrometer (HPLC-HRMS) was employed for analysis. The investigation identified 10 metabolites of roflumilast, including six phase I and four phase II metabolites from in vivo studies, and 11 metabolites from in vitro studies, consisting of eight phase I and three phase II metabolites. The identified biotransformation products encompassed processes such as hydroxylation, chlorine substitution, methylation, N-oxide formation, and even the dissociation of methylenecyclopropane and difluoromethane. Furthermore, the study identified three glucuronic acid and one sulfonic acid conjugated phase II metabolites of the investigated drug candidate. The aforementioned findings contribute to the detection and comprehension of the unauthorized utilization of roflumilast in equestrian sports.

We developed a method for comprehensive urine drug screening by applying dilute-and-shoot extraction and vacuum-insulated probe-heated electrospray ionization with ultra-high performance liquid chromatography high-resolution quadrupole time-of-flight mass spectrometry (DS-UHPLC-VIP-HESI-QTOFMS). The method involved five-fold post-hydrolysis dilution of urine samples and chromatography on a C18 UHPLC column prior to QTOFMS analysis. The recently introduced VIP-HESI ion source was chosen due to its enhanced ionization efficiency and compatibility with UHPLC-QTOFMS. Extensive data was acquired in positive ion mode with a low collision energy (7 eV) and an elevated collision energy (30 eV), using the broadband collision-induced dissociation data acquisition scan mode that continuously generated high-resolution and accurate mass for parent and fragment qualifier ions, and parent ion isotopic patterns. Compound identification was performed against an in-house database with 1263 compound entries, using an automated post-run reverse target database search with preset identification criteria. Method validation with 56 different drugs showed acceptable results for the limit of identification (median 5 ng/mL), matrix effects (70-130%), repeatability of retention times (< 1%), mass accuracy (< 1 mDa), as well as for specificity and stability. As compared with an established UHPLC-QTOFMS method relying on solid-phase extraction and conventional electrospray ionization, DS-UHPLC-VIP-HESI-QTOFMS produced comparable results from authentic clinical urine samples for most drugs, but showed clearly improved detectability for pregabalin, gabapentin, and ritalinic acid. We anticipate that the new method will be a step forward for laboratories performing routine urine drug screening due to its fast turnaround time, reduced manual workload, cost efficiency, and broad substance coverage.