Drug Testing and Analysis最新文献

The enactment of the generic ban on synthetic cannabinoid receptor agonists (SCRAs) in China (2021) added a new flavor to the already diverse and complex SCRA market. Although a large portion of SCRAs is covered by this legislation, a novel strategy to bypass the ban has emerged. So-called “DIY” (do-it-yourself) kits and semi-finished SCRAs are now being offered online, allowing users or intermediate suppliers to purchase ban-evading precursors, with the aim that buyers finish the synthesis. Using in vitro β-arrestin2 recruitment bioassays, we assessed the CB₁ and CB₂ receptor activation potential of three methyl-3,3-dimethyl-butanoate SCRA precursors (MDMB-ICA, MDMB-INACA, and MDMB-5'Me-INACA), along with some of their potential finished end products, including typical, well-known but scheduled SCRAs (e.g., 5F-MDMB-PINACA and 5F-MDMB-PICA), as well as some more recent substances (MDMB-BUTICA). Whereas tail-less precursors were weakly active at CB₁ (EC₅₀ values of 2.34 μM and higher), “finished” SCRAs ((4F-)MDMB-BUTI (NA)CA and (5F-)MDMB-PI (NA)CA) strongly activated CB₁ (EC₅₀ 1.01–35 nM and E_max 366%–488% [relative to JWH-018]). This emphasizes that this “DIY” synthesis phenomenon poses a serious threat to public health, as it is a new indirect way of “legally” providing users with very potent (known) compounds. Importantly, the “DIY” strategy currently ensures the continued presence of scheduled substances on the market, as exemplified by forensic cases from the United States. While precursors can often not be detected because of a concentration below the limit of detection, it is hypothesized that the presence of SCRAs in at least some of these cases stems from this ban-evading strategy.

The athlete biological passport (ABP) has been established as an anti-doping tool based on the statistical analyses of an athlete's biological variables over a period of time. It was introduced in 2007. An important aspect to ensure interlaboratory comparability was to use only one analytical platform—the Sysmex XT-2000. When the new Sysmex XN-1000 platform replaced the XT-2000i in 2019, there was a bias for the reticulocyte percentage. Although clinically insignificant, it interfered with interpreting athletes' haematological profiles for anti-doping purposes; therefore, it was necessary to adjust the haematological module. With the introduction of the new Sysmex XR-Series in 2023, an implementation of this new instrument could become necessary in the future. While the analytical performance of the XR-Series for clinical purposes has been evaluated previously, data in the context of ABP requirements, which are defined in WADA's technical documents, are not available. Therefore, our goals were to compare the XR-series with the XN-1000 and to evaluate their performance within an anti-doping context.

Over 300 samples were analysed on the two instruments following WADA's technical document TD2021BAR, which defines the analytical requirements.

The results for all ABP parameters including the calculated OFF-Score (OFF-hr) and the Abnormal Blood Profile Score (APBS) showed excellent interplatform comparability.

In conclusion, our study demonstrates that the Sysmex XR meets WADA's requirements for haematological analysis. It can confidently replace the Sysmex XN in anti-doping laboratories without compromising the integrity of WADA's ABP longitudinal profiles.

Nitazene analogs are potent novel synthetic opioids (NSOs) that are becoming increasingly common and pose a threat to the public because of their fentanyl-like effects. Although 12 nitazene analogs are currently classified as Schedule I under the U.S. Controlled Substances Act, novel analogs continue to emerge, making their identification in forensic laboratories exceedingly difficult. Liquid chromatography–electrospray ionization–tandem mass spectrometry (LC–ESI–MS/MS) is commonly utilized in toxicology laboratories and is becoming more common for seized drug analysis, particularly for compounds less suited for gas chromatography–electron ionization–mass spectrometry (GC–EI–MS). This study provides a comprehensive structural characterization of 38 representative nitazene analogs using LC–ESI–MS/MS instrumentation, including the proposed fragmentation mechanisms that lead to the formation of diagnostic product ions, enabling analog differentiation. General fragmentation pathways and mechanisms are proposed for all nitazene analogs, including inductive cleavages and molecular rearrangements. Overall, the most common product ions for nitazene analogs are derived from the substitutions to the amine or benzyl moieties, such as m/z 100, m/z 72, m/z 44, and m/z 107. However, the presence of different substitutions shifts the observed product ions. For example, recently occurring piperidine or pyrrolidine rings produce diagnostic product ions at m/z 112 and m/z 98, respectively. Therefore, modification to the core nitazene structure produces different diagnostic product ions, which can be used to identify existing and novel analogs. This study provides a comprehensive assessment of the fragmentation behavior of nitazene analogs under ESI–MS/MS conditions, which provides the basis for identifying new structural modifications in novel nitazene analogs.

Thyroid hormones (TH) are widely used for treatment of hypothyroidism or thyroid cancer in medical practice, but their use among elite athletes without thyroid disease remains controversial. Despite lacking clear ergogenic benefits, some athletes reportedly use thyroxine (T4) and/or triiodothyronine (T3) with the belief that they enhance performance or facilitate weight management. This study investigates self-reported TH use by athletes at the Tokyo 2020, Beijing 2022, and Paris 2024 Olympic Games, analyzing trends based on sex, age, country, and sport. Across these three events, 1.7% of tested athletes reported TH use, with reported use higher among females and in explosive power sports, which is higher than expected for the youthful Olympic participants studied. However, TH use has declined to 1.3% at Paris 2024 compared to the previous Games (Tokyo 2020: 1.8%; Beijing 2022: 2.7%). Whether this decline is due to a growing awareness of the potential harms and/or lack of performance benefits could not be determined by this study.

This paper describes the studies of the in vitro biotransformation of nandrolone decanoate and its metabolic fate in equine plasma and urine after intramuscular administration to castrated thoroughbred horses. The in vitro metabolic study was performed using homogenised horse liver, and the more prominent in vitro biotransformation pathways were found to include hydrolysis, reduction, oxidation and sulfation, mainly resulting in seven Phase I metabolites and one Phase II metabolite. The administration study of nandrolone decanoate was carried out using three retired thoroughbred geldings, each of which was administered intramuscularly with 800 mg of nandrolone decanoate (Deca-Durabolin) once weekly for three consecutive weeks. Nandrolone decanoate and the majority of the identified in vitro metabolites were detectable in post-administration plasma samples for at least 20 days (last sample collected) after the last administration. Although nandrolone decanoate was not found in any post-administration urine, nandrolone (as a metabolite) and its downstream metabolites can be detected for at least 20 days post-administration (last sample collected). For doping control purpose, nandrolone decanoate itself could be a suitable target analyte in horse plasma for effectively controlling its misuse in equine sports.

Although the abuse of muscle-building compounds in elite sports is already known for a long time, these products have become more popular in recreational sport over the past years. Although anabolic steroids are still the most popular ones in this context, the use of other molecules with anabolic properties is on the rise. Three categories of such products are the selective androgen receptor modulators (SARMs), the metabolic modulators and the growth hormone secretagogues (GHS). Based on this trend and the outcomes of a previous market surveillance study in the domain of illegal products (MSSIP), the Falsified Medicines Working Group of the General European Official Medicines Control Laboratories (OMCL) Network (GEON) decided to conduct an MSSIP with focus on SARMs, metabolic modulators and GHS over a period of 5 years. In total 324 samples and 354 results, reported by 14 laboratories in 13 countries, members of the GEON, were taken into account, for which the majority of the samples originated from illegal distribution. Sixty-five percent of the seized products were represented as medicine, though 24% as dietary supplements, which is of concern because here the (recreational) sporter is not aware he is taking an (unapproved) pharmaceutical. Eighteen different molecules, within the scope of the study, were reported with as top 5: ibutamoren, ligandrol, ostarine, cardarine and andarine. From the limited quantitative data reported, it can be assumed that the majority of the samples contain active doses and some are even overdosed, so health risks for the consumers cannot be neglected.

Benzodiazepines and related z-drugs (BZDs) are widely used pharmaceuticals but require careful monitoring due to limited efficacy, tolerance development and the risk of dependence. Wastewater-based epidemiology (WBE) is a valuable approach to gather population-wide information on consumption patterns through analysis of influent wastewater. This study examines how overlapping metabolic pathways within the BZD drug class can lead to misinterpretation if not carefully considered. An analytical method is developed and validated for 26 biomarkers of prescription BZDs in influent wastewater using solid-phase extraction and liquid chromatography–tandem mass spectrometry and applied to influent wastewater samples for the first time in Belgium (13 locations), Latvia (1), Lithuania (3) and Romania (1). The countries were chosen for their differing prescription BZD market registration, allowing testing of different hypotheses. To aid in complex data interpretation, this study presents a structured categorisation of BZDs into direct biomarkers, which directly reflect consumption, and downstream biomarkers, which are influenced by multiple BZDs, and further discusses the implication this has on the interpretation. Spatial differences were noted among direct biomarkers such as higher consumption of zolpidem in Belgium and zopiclone in Riga (LVA). Downstream biomarkers had higher population-normalised mass loads but cannot be interpreted in isolation, for example, lorazepam (ranging 4–25 mg/day/1000 inhabitants over all locations), lormetazepam (8–30), oxazepam (10–50) and temazepam (1–12). These findings highlight challenges in interpreting BZD consumption trends and emphasise the need of distinguishing different biomarker classes to ensure accurate and comparable results across studies needed to guide informed decision-making.

Drug-checking programs use point-of-need testing (e.g., test strips) and laboratory-based analysis to rapidly identify emerging drug threats, but each has limitations. Test strips are quick but compound or class specific, whereas laboratory testing can identify more compounds but have lengthy turnaround times. To address these limitations, it was proposed that compounds could be extracted from used test strips for additional analyses allowing for rapid on-site information followed by comprehensive laboratory results. The method development process involved four parts: determining the optimal extraction approach, assessing the feasibility of performing direct analysis in real-time mass spectrometry (DART-MS) analysis on extracts, determining the limits of detection (LODs) for a range of analytes, and evaluating the method using used test strips submitted by harm reduction sites. The optimized method consisted of extracting analytes of interest from a cut test strip using 0.5-mL methanol while vortexing for 10 s. DART-MS successfully identified the compounds of interests, and the test strip chemical background was identified. LODs were found to be as low as a mass fraction of 0.005 in a mixture. For the samples submitted by harm reduction sites, concordance between extracts and test strip results was 96%, and the agreement in compound identification between used test strip extracts and authentic drug collection samples was approximately 80% regardless of test strip type and preparation. This work shows that additional analyses of extracted test strips can provide a low-barrier way for high-quality testing that can be used to increase data on the drug landscape.

In the past few years, doping tests have been performed with dried blood spots (DBS) as test samples; this sampling method is minimally invasive and requires minimal storage space. Steroid esters are stable in dried blood, allowing the direct analysis of testosterone esters. In Japan, the cream-based formulation containing 17α-methyltestosterone (MeT) and testosterone propionate (TP) is available as an over-the-counter drug for hair growth. Here, we investigated whether the transdermal uptake of this formulation could be detected using the existing method for steroid ester analysis in DBS. A method that was previously validated to detect and identify TP could also detect MeT. We developed a high throughput liquid chromatography–tandem mass spectrometry method to identify MeT in DBS. The limits of detection and identification of MeT were 0.1 ng/mL and 0.6 ng/mL, respectively. Five male human subjects received the cream-based formulation of MeT and TP transdermally. We then collected blood samples by finger pricking and made DBS on cellulose paper. Using this method, MeT was detected and identified up to 97 h after the final application. TP was also detected up to 97 h and identified up to 49 h after the final application. The developed method provides a direct confirmation of the presence of the drug. Moreover, using cream preparations could lead to contamination from MeT and TP remaining on the fingertip skin during fingertip puncture sampling.