Drug Testing and Analysis最新文献

Triptorelin, a synthetic gonadotrophin-releasing hormone (GnRH), is mainly used in the clinical treatment of prostate cancer. The mechanism initially stimulates luteinizing hormone (LH) and testosterone secretion followed by suppression, resulting in a reduction in cancer progression. However, GnRHs are prohibited in doping control because of the indirect surge of LH and testosterone. Therefore, GnRH analog detection and confirmation are enforced by World Anti-Doping Agency (WADA) requirements. The effects of triptorelin on LH and endogenous steroid levels in urine and serum of five prostate cancer patients taking triptorelin for the first time were investigated and compared with leuprorelin. The samples were collected at 0.0 h, 3.0 h, 6.0 h, 1 month, and 3 months later after drug administration. The effect of triptorelin on LH levels was measured using a sandwich enzyme-linked immunoassay (ELISA). Testosterone and endogenous steroid levels were monitored using gas chromatography coupled with mass spectrometry (GC/MS). Triptorelin showed an advantage over leuprorelin on LH and testosterone suppression, which is preferable to use for prostate cancer treatment. In this study, triptorelin (5-10), a unique in vivo metabolite, was found in urine and serum and verified with synthetic triptorelin (5-10). The metabolite was analyzed using liquid chromatography combined with Orbitrap (LC-Orbitrap) and liquid chromatography coupled with ion trap/time-of-flight mass spectrometry (LC/MS-IT-TOF). When triptorelin levels are undetectable, the presence of triptorelin (5-10) in human urine can be used as evidence that triptorelin is being misused in doping control.

Since the early 20th century, the concept of doping was first introduced. To achieve better athletic performance, chemical substances were used. By the mid-20th century, it became gradually recognized that the illegal use of doping substances can seriously endangered athletes' health and compromised the fairness of sports competitions. Over the past 30 years, the World Anti-Doping Agency (WADA) has established corresponding rules and regulations to prohibit athletes from using doping substances or restrict the use of certain drugs, and isotope, chromatography, and mass spectrometry techniques were accredited to detect doping substances. With the development of gene editing technology, many genetic diseases have been effectively treated, but enabled by the same technology, doping has also the potential to pose a threat to sports in the form of gene doping. WADA has explicitly indicated gene doping in the Prohibited List as a prohibited method (M3) and approved qPCR detection. However, gene doping can easily evade detection, if the target genes' upstream regulatory elements are considered, the task became more challenging. Hi-C experiment driven 3D genome technology, through perspectives such as topologically associating domain (TAD) and chromatin loop, provides a more comprehensive and in-depth understanding of gene regulation and expression, thereby better preventing the potential use of 3D genome level gene doping. In this work, we will explore gene doping from a different perspective by analyzing recent studies on gene doping and explore related genes under 3D genome.

Testosterone, nandrolone, and boldenone, which are listed as doping substances on the World Anti-Doping Agency Prohibited List, are mostly available commercially in esterified forms. Isotope ratio mass spectrometry (IRMS) represents a key tool for identifying these substances, as they are hydrolyzed and discharged in the urine as pseudo-endogenous substances. However, IRMS, which comprises a complicated process, cannot achieve the direct detection of steroid esters in blood samples. These substances can be detected using dried blood spots (DBSs), reducing the impact of esterase hydrolysis. Here, a simultaneous liquid chromatography-tandem mass spectrometry method for detecting 28 steroid (13 testosterone, nine nandrolone, and six boldenone) esters was developed using three DBS types of samples, including a cellulose paper and polymer. The substances were first derivatized with methyloxime to increase their sensitivities (the limits of detection were <0.1-0.4, <0.1-0.9, and <0.1-0.9 ng/mL for the testosterone, nandrolone, and boldenone esters, respectively). Further, the DBS absorbents were verified since the effect of interferences depended on it. Next, a study involving seven participants was conducted to detect intramuscularly administered testosterone enanthate (100 mg). Polymer and cellulose papers were used to collect blood from their upper arms and fingertips, respectively, and testosterone enanthate was identified and detectable at both blood-collection sites for up to 144 and 216 h, respectively. Furthermore, testosterone enanthate was detectable in the DBS samples stored under refrigeration after 6 months, indicating the stable nature of DBS.

Estra-4,9-diene-3,17-dione (dienedione) is an anabolic androgenic steroid (AAS) sold as a bodybuilding supplement. It is prohibited in both human and equine sports. With no report of 4,9-diene configuration in endogenous steroids, dienedione has long been considered a synthetic AAS. Nevertheless, the reoccurring detection of dienedione in colt (entire male horse) urine samples lead to the investigation of its possible endogenous nature in horses. This paper describes (i) the detection of naturally occurring dienedione in colts, (ii) the conjugation study of dienedione and (iii) the population study of free and glucuronide-conjugated dienedione in colt urine. Qualitative and quantitative analyses of dienedione content in colt urine were performed, employing liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Qualitative analyses showed that dienedione was endogenous in colt urine and mainly in the form of glucuronide conjugates. Glucuronidation of dienedione was believed to happen at 3-enol leading to dienedione-3-glucuronide. Upon the population study of free and glucuronide-conjugated dienedione in colt urine samples (n = 175), the mean ± SD was determined to be 2.5 ± 3.5 ng/ml. The population data fitted a normal distribution after a fifth root transformation with the exclusion of one outlier by Grubb's test. A possible in-house threshold was proposed at 30 ng/ml of free and glucuronide-conjugated dienedione in colt urine associated with a risk factor of 1 in 14,269 (with a degree of freedom of 173). This is the first report of endogenous dienedione in entire male horses and the approach for controlling its potential misuse by using a threshold is also presented.

Wastewater-based epidemiology was applied in northeastern Brazil during a dance festival, revealing that cocaine consumption doubled during the event days. The daily drug loads were 0.95 ± 0.03 to 11.4 ± 0.4 g/day for BE, 1.8 ± 0.4 to 7.6 ± 0.3 g/day for COC, 0.04 ± 0.02 to 0.19 ± 0.02 g/day for COE, and 0.08 ± 0.02 to 0.80 ± 0.02 g/day for MDMA.

As negative drug tests are frequently a condition for employment, some people who use drugs will try to subvert the testing. In this study, systematic web monitoring was used to investigate how drug test subversion is discussed online. Posts pertaining to drug test subversion were obtained from public websites and the dark web (n = 634, July-December 2021). Most information from public websites came from Twitter (65%), and 94% of dark web posts were from Reddit. The posts were manually coded to extract quantitative and qualitative information about drug test subversion tactics. Most posts discussed urine drug tests (85%), followed by hair (11%) and oral fluid (2%), and the most discussed drugs were marijuana (72%) and cocaine (7.3%). Urine drug test subversion mainly pertained to specimen substitution, with synthetic urine or urine from another person. Another strategy was to mask diluted urine by ingesting creatine. Urine adulteration was rarely discussed. Hair test subversion involved harsh treatments with products such as bleach, baking soda, and/or detergent. Hair removal was also discussed. Oral fluid test subversion focused on removing drugs from the oral cavity through vigorous brushing of teeth and tongue as well as the use of mouthwash, hydrogen peroxide, gum, and commercial detox products. This study highlights subversion strategies used by donors. Although little evidence was provided as to the effectiveness of these strategies, this information may help guide future studies and development of specimen validity testing to minimize the impact of drug test subversion attempts.

The use of performance-enhancing substances not only undermines the core values of sports but also poses significant health risks to athletes. In a fast-evolving doping environment, where sport professionals are constantly seeking novel and illegal means to bypass doping tests, and new substances are regularly detected on the drug market, it is crucial to inform authorities with updated evidence emerging from scientific research. The current study aims to (i) outline the structure of knowledge in the literature on performance enhancers in sports (i.e., most active countries, main sources, most productive authors, and most frequently used keywords); (ii) identify the most impactful documents in the field; and (iii) uncover the main domains of research in the literature. To do so, we conducted a comprehensive scientometric analysis of the literature on doping, sourcing our data from Scopus. Our research involved a document co-citation analysis of 193,076 references, leading to the identification of the 51 most influential documents and seven key thematic areas within the doping literature. Our results indicate that the scientific community has extensively studied the most prevalent doping classes, such as anabolic agents and peptide hormones, and little is still known about the use of contaminated supplements or other types of enhancers identified as emergent trends. Concurrently, technological advancements contributed to the development of more sophisticated doping detection techniques, using blood or urine samples. More recently, the focus has shifted towards the athlete biological passport, with research efforts aimed at identifying biomarkers indicative of doping. The dynamic nature of doping methods underlines the necessity for more robust educational campaigns, aiming at raising awareness among sports professionals and their entourage about the dangers of doping and the intricacies of its control mechanisms.

As gamma-hydroxybutyric acid (GHB) underlies fast metabolization, its determination from hair may presumably offer a detection window superior to that of body fluids. Due to the wide range of endogenous concentration levels, the evidence of an exogenous ingestion is challenging. As already shown for other drugs, the temporal resolution obtained by applying single hair microanalysis provides further information. Therefore, a method for the extraction and quantification of GHB in 2-mm hair segments (seg) was optimized and validated (limit of detection [LOD]: 2.5 pg/seg, lower limit of quantification [LLOQ]: 5 pg/seg), and five single hairs were examined, each for three non-users and for three (alleged) users. A major challenge was the choice of appropriate extraction tubes without remains of GHB. In two samples from non-users, GHB could not or could only be detected in trace amounts. In the third sample, concentrations between the LOD and 31.1 pg/seg (mean: 9.5, median: 8.4; each pg/seg) were detected with decreasing values towards the tips. In two samples of persons with assumed GHB intake, maximum concentrations of 6.8 and 30.7 pg/seg were measured, but no significant concentration peaks indicating a single ingestion could be observed. The third sample showed concentrations of 7.6-55.2 pg/seg (mean: 28.8, median: 29.6; each pg/seg). In this case, the obtained profiles showing at least two reproducible concentration maxima between 20 and 40 mm point to an ingestion of GHB. The concentration profiles from single hairs were reproducible in each case, reflecting the concentration course of routine 1-cm segmental analysis. These are the first results published on GHB testing in segmented single hairs, and the results must be verified further.

Infant exposure to drugs of abuse represents a worldwide problem whose extent is difficult to estimate. Despite the potentially serious health consequences, few data concerning exposure in children under 1 year of age are available. Since in clinical and forensic settings, neonatal and infant hair testing represents a useful method for investigating suspected drug exposures, an observational retrospective study was performed on hair analysis of children under 1 year of age evaluated at the University Hospital of Padova between 2018 and 2022 with the aim of estimate the extent and define the characteristics of this phenomenon in the reference setting. The sample included 102 infants. Chemical-toxicological analyses were requested in 38 cases (37.3%) because of clinically suspicious symptoms of the child (e.g., neuropsychiatric symptoms and suspected neonatal abstinence syndrome) and in 64 cases (62.7%) because of other reasons (e.g., maternal drug history, at-risk environment, and suspected maltreatment). Based on the presence or absence of symptoms in the request, the sample was subdivided into two groups. Hair analysis in these two showed the presence of drug of abuse, respectively, in 44.7% and 67.2% of the cases (p = 0.026). Cocaine was the most frequently detected substance, followed by opiates, and it was detected less frequently in cases investigated for suspicious clinical symptoms (p < 0.05). The results confirm the difficulties in interpreting the clinical picture and in defining the extent of exposure to drugs of abuse. An integrated assessment is fundamental to interpret the case and achieve adequate care of the child.

The 17th edition of the annual banned-substance review on analytical approaches in human sports drug testing is dedicated to literature published between October 2023 and September 2024. As in previous years, focus is put particularly on new or enhanced analytical options in human doping controls as well as investigations into the metabolism and elimination of compounds of interest, which represent central (while not exclusive) cornerstones of the global anti-doping mission. New information published within the past 12 months on established doping agents as well as new potentially relevant substances are reviewed and discussed in the context of the World Anti-Doping Agency's 2024 Prohibited List. Thereby, analytical challenges, especially with regard to the continuously growing number of target compounds and potentially relevant drug classes as well as the exigency (and consequences) of utmost analytical retrospectivity, are thematized and contextualized. Investigations especially into anabolic agents, peptide hormones, and strategies for the detection of gene doping were identified as core areas of anti-doping research in the reviewed period.