Anti-doping Testing Research Articles

Comparison of Microcapillary Blood Sampling Devices for Use in Anti-Doping.

Blood collection is an important facet of anti-doping testing, forming the basis of the hematological module of the athlete biological passport (ABP). Presently, whole blood is collected via venepucture under standardized conditions, in accordance with the World Anti-doping Agency's International Standard for Testing and Investigations (ISTI). Advances in capillary whole blood collection technology now afford the ability to collect microvolumetric capillary whole blood from the upper arm (or other suitable vascular location such as the abdomen) that is "needle-free" and virtually painless. Previous work has demonstrated the suitability and feasibility of capillary whole blood compared with venous using the Tasso+ device. Recently, other microcapillary devices have received FDA approval increasing the options available to the anti-doping community. The present study extends previous work, by collecting microliter capillary whole blood samples via two different FDA approved devices (RedDrop and the Tasso+ device) to compare with venous blood collected simultaneously. Ten participants provided three matched blood samples (2× capillary and 1× venous) in accordance with WADA ABP guidelines for blood sample collection, for a total of 30 samples. Capillary samples from both devices showed excellent laboratory agreement with venous blood for all CBC parameters, with the exception of platelets. Excellent laboratory agreement was also observed between the two microcapillary collection devices. Irrespective of the device, microcapillary blood collection provides a valid alternative to venous blood collection for ABP purposes.

Anabolic-androgenic steroids in sport

Introduction and objective Anabolic-androgenic steroids (AAS) are a group of doping substances. They cause muscle hypertrophy and an increase in strength. Moreover, they may lead to dangerous side effects. Knowledge of the side effects of AAS use is necessary because in clinical practice we often encounter patients who decide to supplement with steroids. The aim of the study is to present information on the epidemiology, mechanism of action, methods of detecting AAS, the impact of the substance on the male and female body, and a description of organ side effects, highlighting: the cardiovascular system, the nervous system and the psyche. Review methods Literature search was conducted in the PubMed database with inclusion criteria ‘free full texts’ and publications date between 2015 – 2024. A total of 7272 results were found; 30 publications were ultimately included in the review. Abbreviated description of the state of knowledge Overusing AAS is very popular among people training at the gym. They have an anabolic effect on muscle tissue. They contribute to infertility in men and masculinization in women. Stimulation of androgen receptors above physiological doses of AAS leads to vascular changes that, in extreme cases, may lead to sudden cardiac death. Moreover, they accelerate brain aging. An increase in aggression and risky sexual behavior has been observed among people using doping. The diagnostic material used in anti-doping tests is urine and blood serum. Summary AAS abuse is a serious public health problem. Men who want to improve their figure and decide to use doping are particularly at risk. Due to the easy availability of steroids, society should be made aware of the negative effects of steroid use.

Antidoping testing and testicle cancer diagnosis: An unexpected helper

Human chorionic gonadotrophin (hCG) is prohibited in male athletes due to its potential performance-enhancing effects. Some athletes with elevated hCG levels during routine doping tests have been diagnosed with testicular cancer, highlighting the inadvertent role of anti-doping screening in cancer detection. This study explores the incidental detection of testicular cancer through routine anti-doping tests among elite athletes, aiming to raise awareness about the potential secondary benefits of these screenings. We analyze cases from the past decade to demonstrate how early detection has facilitated timely medical interventions, allowing athletes to heal and return to competitive sports. Through these instances, athletes become unintentional advocates, contributing to public health awareness about this topic.

Practical steps to develop a transcriptomic test for blood doping

Abstract Blood doping remains a significant problem that threatens the integrity of sport. The current indirect method for detecting blood doping involves the longitudinal measurement of an athlete’s haematological variables and identification of suspicious results that are indicative of doping (i.e., the athlete biological passport). While this has played a significant role in the protection of clean sport, improvements are needed. The development of a transcriptomic test, that can be used to detect the use of blood doping has been discussed and researched for decades and yet, an anti-doping test that can be adopted by the World Anti-Doping Agency (WADA) is yet to be established. With recent advancements in our understanding, as well as in methods of sample collection, the possibility of a transcriptomic test that can be used by WADA, is imminent. There are, however, several practical considerations that must first be made, that will be highlighted in this perspective article.

Breaking Boundaries: Exploring Performance Enhancement and Anti-Doping Testing in Sports.

Breaking Boundaries: Exploring Performance Enhancement and Anti-Doping Testing in Sports.

Beliefs and Practices Levels Associated with Use of Performance Enhancing Substances and Methods among Mixed Martial Arts Athletes in Kenya

Mixed Martial Arts (MMA) has seen a surge in popularity in Kenya, with athletes from various backgrounds participating in the sport. However, with this growth comes an increased risk of performance-enhancing substances (PES) use. The intense physical demands of MMA may drive athletes to seek an edge through these substances, leading to significant concerns about health risks and sports integrity. The aim of this study is to assess the practices and beliefs regarding PES, traditional herbs (TH), and food supplements (FS) among Kenyan MMA athletes. Additionally, the study explores the sources of PES, the factors motivating their use, and the efficacy of current Anti-Doping testing practices. To conduct the study, a cross-sectional survey design was employed, sampling 800 athletes from a target population of 4,000 across six MMA federations. Data were collected using structured questionnaires adapted from the World Anti-Doping Agency (WADA) and the Performance Enhancement Attitude Scale (PEAS), with pre-testing to ensure reliability. Data analysis involved descriptive statistics and One- and Two-Way Analysis of Variance (ANOVA) using SPSS version 22. The results indicate that the majority of MMA participants are male (79.9%), with a median age of 25 years and a mean age of 26.8±4.1 years. Over half of the athletes perceived PES as a problem, but less than one-third viewed traditional herbs as a concern. Food supplements were used by nearly a fifth of the athletes, with bodybuilders showing the highest usage rate (44.4%). Coaches, team doctors, and physiotherapists were primary sources of these substances, while some athletes reported chemists and medical practitioners as suppliers of PES. The study found that the primary reasons for using PES and FS were to boost economic status through prize money, with additional pressures from coaches, family, and colleagues. Anti-doping testing revealed that 20.6% of athletes had been tested out of competition, while 34.4% believed they could provide a sample that wasn't theirs. The study concludes that while many MMA athletes recognize the risks associated with PES, there is a lack of consistent understanding regarding traditional herbs and food supplements. This inconsistency, combined with external pressures, may lead to risky behavior, compromising the athletes' health and the sport's integrity. To address these issues, the study recommends implementing comprehensive education programs for MMA athletes, coaches, and support personnel to raise awareness of PES risks. Strengthening anti-doping regulations and monitoring sources of PES and FS is also crucial. Additionally, promoting ethical coaching practices and addressing external pressures can foster a safer environment and uphold the values of fair competition in Kenyan MMA.

New statistical framework for interlaboratory evaluation of anti-doping testing results by WADA

The World Anti-doping Agency (WADA) International Standard for Laboratories (ISL), developed as part of the World Anti-Doping Program, requires satisfactory laboratory performance in the WADA External Quality Assessment Scheme (EQAS) in order to obtain and maintain WADA accreditation. Under this mandate, WADA regularly distributes urine and blood test samples to anti-doping laboratories to continuously monitor their proficiency. Over the years, WADA has employed classical, generic statistical methods, in accordance to ISO 13528, to evaluate quantitative EQAS results. Here, we set out the rationale for a modern statistical approach that recognizes and addresses the particular features of the measurement results typically obtained in such tests and present an approach involving Bayesian measurement models and statistical data analysis that is tailored specifically to anti-doping testing.

Excessive androgen exposure and risk of malignancies: A cohort study.

A link between androgen use and the risk of cancers, especially prostate and breast cancer, has been suggested. The knowledge about a possible association is limited. The study aimed to investigate cancer incidence rates, particularly those related to prostate and breast cancer, in male androgen users and compare them to a control group. We included male androgen users identified through a nationwide anti-doping testing program in Danish fitness centers from 2006 to 2018. We paired each case with 50 male controls of the same age, selected randomly. The cohort was followed from baseline and until 2023. The outcome was the incidence of prostate cancer, breast cancer, or any cancer excluding non-melanoma skin cancer. The study included 1,189 androgen users and 59,450 controls, with a mean age of 27 years at enrolment. During the follow-up period with a mean length of 11 years, 13 androgen users, and 612 controls were diagnosed with cancer. This resulted in an incidence rate ratio of 1.05 (95% CI: 0.55-1.81). None of the androgen users were diagnosed with prostate or breast cancer. Male androgen users did not face an increased short-term risk of cancer, neither overall nor related to prostate or breast cancer. Our study indicates that the absolute risk of malignancies in androgen users is comparable to that in the background population. However, we cannot exclude androgens as a cancer risk factor due to the limited sample size, relatively short follow-up period, and subject age.

Anti-doping and National Politics: An Ethnography in Brazilian Anti-doping Around the Era of the Rio de Janeiro Games

Abstract This study is based on an investigation into the Brazilian anti-doping policy, with a specific focus on the establishment and operational mechanisms of the Brazilian Doping Control Authority (ABCD), under the auspices of the Ministry of Sport (ME). The objective was to describe how Brazilian national policy (general political decisions, interests of parties or particular politicians/agents) affects the fight against doping and, vice versa, how the harmonization process controlled by the World Anti-Doping Agency (WADA) affects national sports policy. A multi-sited ethnographic study was undertaken, involving field diary practices, interviews and document analysis. Adopting the perspective of pragmatic sociology, this study delves into the descriptions of three key periods in the recent history of anti-doping in Brazil: the creation of ABCD and the dissolution of ANAD (National Anti-Doping Agency), the realization of the first 100 anti-doping tests by the ABCD (critical period for its members who managed to keep away ex-officers from ANAD) and the impeachment of the Brazilian president in 2016 (ex-officers from ANAD took over the direction of the ABCD). Our analysis allowed us to conclude that ‘diverting others from decision-making spheres’ comprised a political strategy to occupy spaces of power or to stabilize itself in them, which, in this case, allowed the control of anti-doping actions in the country. Local protagonists tried to make use of international injunctions and pressure or local political events to achieve these goals.

Detection of doping substances in paired dried blood spots and urine samples collected during doping controls in Danish fitness centers.

The use of dried blood spot (DBS) in anti-doping can be advantageous in terms of collection, transportation, and storage compared with the traditional anti-doping testing matrices urine and venous blood. There could, nonetheless, be disadvantages such as shorter detection windows for some substances compared with urine, but real-life comparison of the detectability of prohibited substances in DBS and urine is lacking. Herein, we present a liquid chromatography-high resolution mass spectrometry (LC-HRMS)-based screening method for simultaneous detection of 19 target analytes from the doping substance categories S1-S5 in a single spot. Ninety-eight urine and upper-arm DBS (Tasso-M20) sample pairs were collected from fitness centers customers notified for doping control by Anti Doping Denmark, and three sample pairs were collected from active steroid users undergoing clinical evaluation and treatment at a Danish hospital. The analytical findings were cross compared to evaluate the applicability of the developed DBS testing menu in terms of feasibility and analytical performance. To our knowledge, this is the first study to compare the detectability of prohibited substances in DBS and urine samples collected in a doping control setting. Twenty-seven of the urine samples and 23 DBS samples were positive, and we observed a very high concordance (95%) in the overall analytical results (i.e., positive or negative samples for both urine and DBS). Collectively, these results are very promising, and DBS seems suitable as a stand-alone matrix in doping control in fitness centers likely because of the high analyte concentration levels in these samples.

Routine application of SFC-MS in doping control: Analysis of 3 × 1000 urine samples using three different SFC-MS instruments.

Supercritical fluid chromatography-mass spectrometry (SFC-MS) has proved to be a beneficial tool for sample analysis for a wide variety of compounds and, as such, has recently gained the attention of the anti-doping community. We have tested the applicability of SFC-MS for routine doping control analysing approximately 3 × 1000 identical anti-doping samples utilising SFC-MS instruments from three different vendors: Agilent Technologies, Waters Corporation and Shimadzu Corporation. A 'dilute and inject' approach either without or after hydrolysis of glucuronide metabolites was applied. Most of the compounds included in our study demonstrated excellent chromatography, whereas some showed co-elution with endogenous interferences requiring MS discrimination. Retention times typically were very stable within batches (%CV ≤ 0.5%), although this appeared to be analyte and column dependent. Chromatographic peak shape was good (symmetrical) and stable over the period of the testing without any change of column. Our results suggest that SFC-MS is a sensitive, reproducible and robust analytical tool ready to be used in anti-doping laboratories alongside the currently applied techniques such as gas and liquid chromatography coupled to mass spectrometry. Even if instruments are designed slightly differently, all three setups demonstrated their fitness for the purpose in anti-doping testing.

Application of micro capillary blood sampling in an anti-doping setting.

Blood collection is an important facet of anti-doping testing, forming the basis of the Athlete Biological Passport (ABP). Traditional blood collection via venipuncture can be uncomfortable for athletes, especially those who are tested frequently or close to competition. Athletes may also have negative perceptions of venipuncture due to past experiences or the risks of adverse health events such as bruising, hematomas, syncope, and general discomfort that has the potential to affect performance. Advances in capillary whole blood collection technology now affords the ability to collect micro-volumetric capillary whole blood from the upper arm (or other suitable vascular location such as the abdomen) that is "needle-free" and virtually painless using devices such as the Tasso+. The present study extends previous work, by collecting microliter capillary whole blood samples via the Tasso+ EDTA device in an official anti-doping setting prior to competition, as well as requiring temperature-monitored cold chain shipping by air to the laboratory before analysis. Fifty-eight matched capillary and venous blood samples were collected under official doping control conditions by certified Doping Control Officers. No impact of sample shipment by air under cool conditions was observed on sample integrity. Provided that no visible clots were identified prior to analysis, capillary and venous blood samples showed excellent laboratory agreement for all CBC parameters, with the exception of platelets. Micro capillary blood collection provides a valid alternative to venous blood collection for ABP purposes, with the advantage of providing a more athlete-friendly experience, particularly close to competition.

The association between subjective anti-doping knowledge and objective knowledge among Japanese university athletes: a cross-sectional study.

This study aimed to assess the association between subjective anti-doping knowledge (subjective ADK) and objective anti-doping knowledge (objective ADK) among Japanese university athletes, framed within the context of the Theory of Planned Behavior (TPB). Eligible participants were 486 university athletes [320 men (65.8%), 166 women; mean age of 18.9 ± 1.0 years]. The participants categorized themselves in terms of the quality of their anti-doping knowledge. This assessment resulted in an independent variable coded as "(1) substantial lack of adequate knowledge," "(2) some lack of adequate knowledge," "(3) fair amount of knowledge" or "(4) good amount of knowledge." Objective ADK was assessed using the Athlete Learning Program about Health and Anti-Doping (ALPHA) test, a set of questions derived from the ALPHA-a former World Anti-Doping Agency e-learning program. The test comprises 12 questions (four choices each; passing index: ≧10 points or 80% correct answer rate). ANCOVA was conducted using subjective ADK as an independent variable and ALPHA scores as a dependent variable, adjusting for confounding factors (anti-doping experience). The ALPHA corrected answer rate across subjective ADK levels for the group were 73.10% for "(1) substantial lack of adequate knowledge," 71.97% for "(2) some lack of adequate knowledge," 75.18% for "(3) fair amount of knowledge" and 72.86% for "(4) good amount of knowledge." Comparison between different levels of subjective ADK revealed no significant differences in ALPHA score considering the main effects or any of their interactions. The present results revealed that Japanese university athletes' subjective ADK did not match their objective ADK. In the context of the TPB, there may be limitations in the perceived behavioral control in anti-doping knowledge. Even if athletes view doping as a wrongful act and have formed attitudes and subjective norms to comply with the rules, the results suggest that errors may occur in the composition of behavioral intentions due to a lack of knowledge. This could lead to the possibility of facing the risk of unintentional anti-doping rule violations. It highlights the need for targeted educational interventions to align subjective ADK of athletes with their objective ADK.

Determination of chlorphenesin in cosmetics, a preservative that can affect anti-doping test result interpretations.

Chlorphenesin is a legitimate preservative commonly used in cosmetics. It shares one urinary metabolite of 4-chlorophenoxyacetic acid with meclofenoxate, a prohibited stimulant in sports. Recently, there have been cases where athletes using chlorphenesin-containing products were falsely identified as users of meclofenoxate. This study developed and validated a liquid chromatography method with diode-array detection to determine the chlorphenesin content in 61 selected personal care products with various functions (e.g., facial care, body cleansing, sun protection, make-up, hairstyling, perfume, and oral cleaning). The analytical method demonstrated fit-for-purpose quantitation and provided good linearity, precision, accuracy, and recovery for analyzing different cosmetic matrices. Among the 27 cosmetics labeled with chlorphenesin, the chlorphenesin concentrations ranged from 0.10 to 2.67 mg/g, with three products showing no detection. None of the products exceeded the maximum limit of 3mg/g (0.3%) set by regulatory authorities. Among the 34 cosmetics not labeled with chlorphenesin, none of them contained chlorphenesin. This study confirmed the absence of undeclared chlorphenesin in the selected cosmetics, supporting the correctness of chlorphenesin labeling in cosmetics sold in Taiwan. Further investigations studying urinary excretion patterns after different types, doses, frequencies, and sites of cosmetics applications could contribute to strengthen current testing approaches in anti-doping.

Summer Olympic sports and female athletes: comparison of anti-doping collections and prohibited substances detected in Australia and New Zealand vs. France.

Like any athlete, female athletes may be tempted to use prohibited substances during competition or training to enhance their performance. Anti-doping tests performed on female athletes in summer Olympic sports from two geographical areas: Australia/ New Zealand, and France were compared. First, the distribution of sample collections across different sports disciplines, as well as the distribution of substances was investigated. Then the distribution of collections and substances detected in the five sports categories (Strength/Speed, Endurance, Mixed, Motor Skills with High Energy Expenditure, and Motor Skills with Low Energy Expenditure) were studied with consideration of therapeutic use exemptions obtained by the athlete. Australia/New Zealand and France were similar in their overall number of anti-doping collections performed. Likewise, both regions had the same sports disciplines (athletics, aquatics, cycling) and sport categories (Mixed and Endurance) as having the highest number of sample collections. The Motor Skills with High Energy Expenditure, and Motor Skills with Low Energy Expenditure categories had the lowest number of sample collections. However, the number of substances detected was significantly different (p < 0.05) with a greater number of substances found in the French data. There were a few substances in common between the two geographical areas, namely prednisone/prednisolone, carboxy-THC, terbutaline, vilanterol and methylphenidate, but most were different. In-competition tests were the category where most of the AAFs were found.

Risk of Central Serous Chorioretinopathy in Male Androgen Abusers.

Male gender is an important risk factor of central serous chorioretinopathy (CSC), and studies have explored the pathophysiological role of androgens in CSC with conflicting results. In this study, we shed light on this hot topic by exploring the risk of CSC in a large cohort of male androgen abusers. This study included male androgen abusers identified through a nationwide anti-doping test program across Danish fitness centers from January 3 2006 to March 1 2018. For each case, we randomly sampled ten male controls using Danish nationwide registries. These controls were matched in age and date. Cases and controls were followed until May 16 2018. Data on diagnoses were extracted using the Danish National Registry of Patients using ICD-10 codes to identify cases with CSC. We included 1189 cases and 11,890 controls. Mean age at the time of doping sentence was 27.4 ± 6.9years, and mean length of follow-up was 15.8 ± 3.6years. We identified no cases of CSC in androgen abusers, and five cases of CSC in the control cohort. The difference between groups was not statistically significant (P = 1.0). Male androgen abusers were not at increased risk of CSC. Considering the lack of any signal in this large study, we speculate that if male androgen plays any direct role in the pathophysiology of CSC, its role may be subtle at best.

Anabolic-androgenic steroids: Considerations for forensic psychiatry, sports psychiatry, and the law

Anabolic-androgenic steroids: Considerations for forensic psychiatry, sports psychiatry, and the law

Higenamine, Anti-Doping, and Plant-Based Cuisine: A Legal Analysis of Higenamine in Sport Anti-Doping Systems

Current anti-doping policy seeks to protect honest athletes from biochemically overeducated colleagues. However, there is a question of whether the present policy has gone too far. This article illustrates the ambiguity of the anti-doping policy in the context of a particular plant-based substance (i.e., higenamine) by providing certain case studies. In such cases, the process of proof requires the continuous checking of suppositions since an athlete must establish how the prohibited substance could have entered his or her body. This obligation implies that an athlete and the defending team must have legal, medical, dietary, and biochemical knowledge. However, even with all precautions, it is still possible to fail an anti-doping test and be severely punished because it is almost impossible to trace all the sources that caused the prohibited substance to enter the athlete’s body.

Comparison of different approaches for direct coupling of solid-phase microextraction to mass spectrometry for drugs of abuse analysis in plasma

The direct coupling of solid-phase microextraction (SPME) to mass spectrometry (MS) (SPME-MS) has proven to be an effective method for the fast screening and quantitative analysis of compounds in complex matrices such as blood and plasma. In recent years, our lab has developed three novel SPME-MS techniques: SPME-microfluidic open interface-MS (SPME-MOI-MS), coated blade spray-MS (CBS-MS), and SPME-probe electrospray ionization-MS (SPME-PESI-MS). The fast and high-throughput nature of these SPME-MS technologies makes them attractive options for point-of-care analysis and anti-doping testing. However, all these three techniques utilize different SPME geometries and were tested with different MS instruments. Lack of comparative data makes it difficult to determine which of these methodologies is the best option for any given application. This work fills this gap by making a comprehensive comparison of these three technologies with different SPME devices including SPME fibers, CBS blades, and SPME-PESI probes and SPME-liquid chromatography-MS (SPME-LC-MS) for the analysis of drugs of abuse using the same MS instrument. Furthermore, for the first time, we developed different desorption chambers for MOI-MS for coupling with SPME fibers, CBS blades, and SPME-PESI probes, thus illustrating the universality of this approach. In total, eight analytical methods were developed, with the experimental data showing that all the SPME-based methods provided good analytical performance with R2 of linearities larger than 0.9925, accuracies between 81% and 118%, and good precision with an RSD% ≤ 13%.

Presence of higenamine in beetroot containing 'foodstuffs' and the implication for WADA-relevant anti-doping testing.

Higenamine is an alkaloid found within plant species including some that are used in traditional Asian and Chinese herbal medicines. Identified as having mixed mode adrenergic receptor activity, higenamine is present within some nutritional supplements marketed for stimulant and/or weight loss. Its inclusion within nutritional supplements can be via its natural presence within botanical ingredients or as a synthetic additive, often added in mg amounts. The World Anti-doping Agency (WADA) prohibited list has contained higenamine since 2017 as banned at all times in the beta-2 agonist (S3) category, with a reporting level of 10ng/ml for the free parent form in urine. In this study, an investigation into the content of beetroot or beetroot-containing foodstuffs and supplement products was conducted. Higenamine was confirmed as present within the majority of foodstuffs and supplements, with experimental evidence that higenamine can arise within beetroot extracts through heating. The results in this paper demonstrate the first reported evidence of a link between beetroot and this WADA prohibited substance. To investigate the link between intake and excretion, concentrated beetroot drinks were consumed by six individuals and higenamine quantified in their urine. Free higenamine was detected in the urine of all individuals, with maximum measured concentration in samples of less than 1% of the current WADA reporting limit. Although the risk of an inadvertent doping violation by consumption of the foodstuffs and products investigated in this study is low, beetroot as a source of higenamine should be considered by athletes.