No pain, just gain: Painless, easy, and fast dried blood spot collection from fingertip and upper arm in doping control

3 Aug 2021

No pain, just gain : Painless, easy, and fast dried blood spot collection from fingertip and upper arm in doping control / Sara Amalie Solheim, Thomas Kamm Ringsted, Nikolai B. Nordsborg, Yvette Dehnes, Maren Christin Stillesby Levernaes, Jakob Mørkeberg. - (Drug Testing and Analysis (2021) 3 August; 3135)

  • PMID: 34346172
  • DOI: 10.1002/dta.3135


Abstract

This study aimed to determine and compare the perception, painfulness, and usability of the minimally invasive dried blood spot (DBS) collections from fingertip versus upper arm from different athlete populations: males and females representing sports dependent on hand/arm, sports less dependent on hand/arm and para-athletes. To accomplish this, 108 national level athletes from Denmark were recruited (♀ = 49, ♂ = 59, 25 ± 6 years; mean ± SD) and 11 Doping Control Officers (DCOs) collected manual fingerprick DBS (HemaSpot HF) and automated upper-arm DBS (Tasso-M20) from each athlete. Athletes and DCOs responded to questionnaires regarding the perception of sample collection procedures. On a 0-10 scale, the athletes reported a low pain score and a very good general experience for both sampling sites, but following upper-arm DBS collection, the associated pain was rated lower (-0.4 ± 1.6, p < 0.05), and the general experience rated better (+0.6 ± 2.3, p ≤ 0.001) than after the fingerprick DBS collection. The DCOs rated the general experience with the upper-arm DBS collection better (+1.6 ± 1.1, p ≤ 0.01) than the fingerprick DBS collection, partly because problems occurred more frequently during the DBS collection from the fingertip (28%) than from the upper arm (6%). In conclusion, it appears that DBS sampling is affiliated with minimal sensation of pain and is preferred by both DCOs and athletes, independent of gender and discipline, over conventional sample collection methods. Collection of DBS from the upper arm was preferred over fingerprick by both athletes and DCOs.

Substance Use and Its Impact on Athlete Health and Performance

7 Aug 2021

Substance Use and Its Impact on Athlete Health and Performance / Todd Stull, Eric Morse, David R. McDuff. - (Psychiatric Clinics of North America 44 (2021) 3 (September); p. 405-417)

  • PMID: 34372997
  • DOI: 10.1016/j.psc.2021.04.006


Abstract

The pressure to gain mass, power, explosiveness, and endurance and to obtain a performance edge continues to a part of sports. Anabolic agents, including selective androgen receptor modulators along with peptides, hormones, and metabolic modulators, continues to evolve. Methods to promote transcription to modify gene expression are a part of the evolution. In order to monitor and improve doping detection, the Athlete Biological Passport has been created. This article provides an up-to-date review of alcohol, anabolic androgens and related agents, stimulants, opioids, and cannabis and related compounds and their effects on athlete health and performance.

Mass spectrometric characterization of urinary hydrafinil metabolites for routine doping control purposes

10 Aug 2021

Mass spectrometric characterization of urinary hydrafinil metabolites for routine doping control purposes / Andre Knoop, Gregor Fußhöller, Nadine Haenelt, Christian Goergens, Sven Guddat, Hans Geyer, Mario Thevis. - (Drug Testing and Analysis (2021) 10 August; 3137)

  • PMID: 34378339
  • DOI: 10.1002/dta.3137


Abstract

Little information on the human metabolism and urinary elimination of hydrafinil (9-fluorenol) exists. In order to support preventive anti-doping activities concerning compounds such as hydrafinil, a pilot elimination study was conducted with three healthy male volunteers receiving a single oral dose of 50 mg of hydrafinil. Urine samples were collected prior to and up to 72-h post-administration and were subjected to both gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry, which allowed for the identification of the intact drug as well as Phase I and Phase II metabolites, primarily hydroxylated and/or glucuronidated or sulfo-conjugated hydrafinil. The identity of these metabolites was corroborated by high-resolution/high-accuracy tandem mass spectrometry, and the applicability of routine doping control workflows for the detection of hydrafinil and its main metabolites was assessed. Therefore, two findings of hydrafinil and its metabolites were recorded, which concerned out-of-competition doping control samples and, hence, were not pursued with confirmatory analyses. Yet, the initial testing procedure results indicate that hydrafinil might require consideration in sports drug testing programs to ensure its detection, if classified as prohibited by the World Anti-Doping Agency (WADA).

A fast screening method for the detection of CERA in dried blood spots

11 Aug 2021

A fast screening method for the detection of CERA in dried blood spots / Angela Rocca, Laurent Martin, Tiia Kuuranne, Magnus Ericsson, Alexandre Marchand, Nicolas Leuenberger. - (Drug Testing and Analysis (2021) 11 August); p. 1-6)

  • PMID: 34380180
  • DOI: 10.1002/dta.3142


Abstract

Continuous erythropoietin receptor activator (CERA) is a third-generation erythropoiesis-stimulating agent that was developed for the treatment of anemia. However, misuse of CERA for doping in endurance sports has been reported. Previous studies have shown blood as the matrix of choice for the detection of CERA, due to its high molecular weight. The use of dried blood spots (DBSs) for anti-doping purposes constitutes a complementary approach to the standard urine and venous blood matrices and could facilitate sample collection and increase the number of blood samples available for analysis due to reduced costs of sample collection and transport. Here, we investigated whether CERA could be indirectly detected in extracts of single DBSs using an erythropoietin-specific immunoassay that is capable of providing results within approximately 2 h. Reconstituted DBS samples were prepared from mixtures of red blood cell pellets and serum samples. The samples were collected in a previous clinical study in which six healthy volunteers were injected with a single, 200 μg dose of CERA. Using a commercially available ELISA kit, CERA was detected in the DBSs with a detection window of up to 20 days post-injection. Furthermore, in order to demonstrate the fitness-for-purpose, three authentic doping control serum samples, which were identified as containing CERA, were analyzed by the presented methodological approach on DBS. The testing procedure described here could be used as a fast and cost-effective method for the detection of CERA abuse in sport.

Pitch-side Acute Severe Pain Management Decisions in European Elite football

16 Aug 2021

Pitch-side Acute Severe Pain Management Decisions in European Elite football / Maeve Claire Doheny, Gerard Bury. - (International Journal of Sports Medicine (2021) 9 (16 August))

  • PMID: 34399429
  • DOI: 10.1055/a-1588-7931


Abstract

This is the first study on acute severe pain management involving sport and exercise medicine Doctors who are leaders in football medicine in their respective countries. An online survey was designed describing the management of acute severe pain in this expert cohort. The survey captured participant sex, age, years working in sports medicine, core specialty and use of clinical practice guidelines (CPGs). Finally, three clinical vignettes exploring the management of acute pain were presented. Forty-four senior team doctors across 55 European countries completed the survey. There were no consistent guidelines proposed, with 33 (75%) participants indicating they did not use any. Methoxyflurane was proposed by 14 (32%) and 13 (30%) participants for female anterior cruciate ligament rupture and male ankle fracture, respectively. Strong opioids were not used in 17 (39%) and 6 (14%) participants regarding female cruciate injuries and male fractures, respectively. Despite 75% of participants having paediatric life support training, eight (18%) participants expressed uncertainty administering medications in this population, and 15 (34%) would avoid using strong opioids altogether. There is a tendency to undertreat pain and avoid strong opioids for reasons including lack of monitoring equipment, anti-doping concerns and lack of comfort treating paediatric patients with opioids.

Controlled Administration of Dehydrochloromethyltestosterone in Humans: Urinary Excretion and Long-Term Detection of Metabolites for Anti-Doping Purpose

19 Aug 2021

Controlled Administration of Dehydrochloromethyltestosterone in Humans: Urinary Excretion and Long-Term Detection of Metabolites for Anti-Doping Purpose / Steffen Loke, Xavier de la Torre, Michele Iannone, Giuseppe La Piana, Nils Schlörer, Francesco Botrè, Matthias Bureik, Maria Kristina Parr. - (The Journal of Steroid Biochemistry and Molecular Biology (2021) 19 August; 105978)

  • PMID: 34418529
  • DOI: 10.1016/j.jsbmb.2021.105978


Abstract

Dehydrochloromethyltestosterone (DHCMT) is an anabolic-androgenic steroid that was developed by Jenapharm in the 1960s and was marketed as Oral Turinabol®. It is prohibited in sports at all times; nevertheless, there are several findings by anti-doping laboratories every year. New long-term metabolites have been proposed in 2011/12, which resulted in adverse analytical findings in retests of the Olympic games of 2008 and 2012. However, no controlled administration trial monitoring these long-term metabolites was reported until now. In this study, DHCMT (5 mg, p.o.) was administered to five healthy male volunteers and their urine samples were collected for a total of 60 days. The unconjugated and the glucuronidated fraction were analyzed separately by gas chromatography coupled to tandem mass spectrometry. The formation of the described long-term metabolites was verified, and their excretion monitored in detail. Due to interindividual differences there were several varieties in the excretion profiles among the volunteers. The metabolite M3, which has a fully reduced A-ring and modified D-ring structure, was identified by comparison with reference material as 4α-chloro-17β-hydroxymethyl-17α-methyl-18-nor-5α-androstan-13-en-3α-ol. It was found to be suitable as long-term marker for the intake of DHCMT in four of the volunteers. In one of the volunteers, it was detectable for 45 days after single oral dose administration. However, in two of the volunteers M5 (already published as long-term metabolite in the 1990s) showed longer detection windows. In one volunteer M3 was undetectable but another metabolite, M2, was found as the longest detectable metabolite. The last sample clearly identified as positive was collected between 9.9 and 44.9 days. Furthermore, the metabolite epiM4 (partially reduced A-ring and a modified D-ring structure which is epimerized in position 17 compared to M3) was identified in the urine of all volunteers with the help of chemically synthesized reference as 4-chloro-17α-hydroxymethyl-17β-methyl-18-nor-androsta-4,13-dien-3β-ol. It may serve as additional confirmatory metabolite. It is highly recommended to screen for all known metabolites in both fractions, glucuronidated and unconjugated, to improve identification of cheating athletes. This study also offers some deeper insights into the metabolism of DHCMT and of 17α-methyl steroids in general.

Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector

16 Aug 2021

Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector / Takehito Sugasawa, Takuro Nakano, Shin-Ichiro Fujita, Yuki Matsumoto, Genki Ishihara, Kai Aoki, Koki Yanazawa, Seiko Ono, Shinsuke Tamai, Lev Manevich, Haruna Ueda, Noriyo Ishibashi, Kenshirou Tamai, Yasuharu Kanki, Yasuko Yoshida, Koichi Watanabe, Tohru Takemasa, Yasushi Kawakami, Kazuhiro Takekoshi. - (Genes 12 (2021) 8 (16 August); 1249)

  • PMID: 34440425
  • PMCID: PMC8392868
  • DOI: 10.3390/genes12081249


Abstract

Despite the World Anti-Doping Agency (WADA) ban on gene doping in the context of advancements in gene therapy, the risk of EPO gene-based doping among athletes is still present. To address this and similar risks, gene-doping tests are being developed in doping control laboratories worldwide. In this regard, the present study was performed with two objectives: to develop a robust gene-doping mouse model with the human EPO gene (hEPO) transferred using recombinant adenovirus (rAdV) as a vector and to develop a detection method to identify gene doping by using this model. The rAdV including the hEPO gene was injected intravenously to transfer the gene to the liver. After injection, the mice showed significantly increased whole-blood red blood cell counts and increased expression of hematopoietic marker genes in the spleen, indicating successful development of the gene-doping model. Next, direct and potentially indirect proof of gene doping were evaluated in whole-blood DNA and RNA by using a quantitative PCR assay and RNA sequencing. Proof of doping could be detected in DNA and RNA samples from one drop of whole blood for approximately a month; furthermore, the overall RNA expression profiles showed significant changes, allowing advanced detection of hEPO gene doping.

Meldonium residues in milk: A possible scenario for inadvertent doping in sports?

10 Aug 2021

Meldonium residues in milk : A possible scenario for inadvertent doping in sports? / Sven Guddat, Christian Görgens, Tim Sobolevsky, Mario Thevis. - (Drug Testing and Analysis (2021) 26 August)

  • PMID: 34448364
  • DOI: 10.1002/dta.3145


Abstract

Lately, the veterinary drug Emidonol® has been discussed as a possible scenario for inadvertent doping in sports. Emidonol® is approved for use in livestock breeding, exhibiting antihypoxic and weak sedative effects. The veterinary drug rapidly dissociates into meldonium, a substance prohibited in sports, and is excreted largely in its unchanged form into urine. To investigate if residues of meldonium in edible produce may result in adverse analytical findings in sports drug testing, a pilot study was conducted with three volunteers consuming a single dose of 100 ml meldonium-spiked milk at a concentration of 500 ng/ml (Study 1), and multiple doses of 100 ml of meldonium-spiked milk (500 ng/ml) on five consecutive days (Study 2). In the single dose study, urinary meldonium concentrations peaked between 2 and 6 h post-administration with maximum values of 7.5 ng/ml, whereas maximum meldonium concentrations of 18.6 ng/ml were determined after multiple doses 4 h post-administration. All samples were analyzed using an established and validated protocol based on HILIC-HRMS/MS.

Coherence of drug policy in sports: Illicit inclusions and illegal inconsistencies

13 Jun 2013

Coherence of drug policy in sports : Illicit inclusions and
illegal inconsistencies / Kathryn Hennea, Benjamin Koh, Vanessa McDermott. - (Performance Enhancement & Health 2 (2013) 2 (June); p. 48-55)

  • DOI: 10.1016/j.peh.2013.05.003


Abstract:

Anti-doping regulation has become a critical element of contemporary sport, conveying what substances and methods athletes may use. Although this form of regulation is often synonymous with banning performance enhancing substances and methods, the World Anti-Doping Agency (WADA), the global authority tasked with formulating policies on drug use in sports also regulates illicit drugs, including those considered non-performance enhancing. This paper examines how the inclusion of illicit drugs on the WADA Prohibited List falls under the agency's mandate by focusing on how the agency justifies the prohibition of substances more generally. In so doing, it highlights inconsistencies in the criteria used to evaluate whether or not substances should be prohibited. After critically considering WADA's criteria, this paper posits a way to reconcile the inconsistencies. It suggests that aligning the criteria with existing claims that doping is a broader health concern provides a way for WADA to clarify ambiguities around why substances are on the WADA Prohibited List and better communicate why it regulates illicit non-performance enhancing drugs.

To dope or not to dope: Elite athletes’ perceptions of dopingdeterrents and incentives

5 Aug 2013

To dope or not to dope : Elite athletes’ perceptions of dopingdeterrents and incentives / Marie Overbye, Mette Lykke Knudsen, Gertrud Pfister. - (Performance Enhancement & Health 2 (2013) 3 (September); p. 119-134)

  • DOI: 10.1016/j.peh.2013.07.001


Abstract:

Aim

This study aims to examine the circumstances which athletes say affect their (hypothetical) considerations of whether to dope or not and explore the differences between athletes of different gender, age and sport type.

Methods

645 elite athletes (mean age: 22.12; response rate: 43%) representing 40 sports completed a web-based questionnaire. Participants were asked to imagine themselves in a situation in which they had to decide whether to dope or not to dope and then evaluate how different circumstances would affect their decisions.

Results

Multiple circumstances had an effect on athletes’ hypothetical decisions. The most effective deterrents were related to legal and social sanctions, side-effects and moral considerations. Female athletes and younger athletes evaluated more reasons as deterrents than older, male athletes. When confronted with incentives to dope, the type of sport was often a more decisive factor. Top incentives were related to qualified medical assistance, improved health or faster recovery from injury, the low risk of being caught and the threat posed to an elite career.

Conclusions

Our results reveal that numerous circumstances affect athletes’ thoughts on doping and athletes of different gender, age and sport type reacted differently to a variety of circumstances that may potentially deter or trigger doping. Particularly notable findings were the potential role of doctors in athletes’ doping and that the current punitive anti-doping approach seems to deter athletes, although the fear of social sanctions was almost as great a deterrent.

Implications

Anti-doping prevention strategies should be diversified to target specific groups of athletes.

KeywordsAnti-doping policyDopingElite athletesGenderSports medicine

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