Annual banned‐substance review: Analytical approaches in human sports drug testing

Higenamine (Norcoclaurine) is a very popular substance in Chinese medicine and is present in many plants. The substance may be also found in supplements or nutrients, consumption of which may result in violation of anti-doping rules. Higenamine is prohibited in sport at all times and included in Class S3 (β-2-agonists) of the World Anti-Doping Agency (WADA) 2017 Prohibited List. The presence of higenamine in urine samples at concentrations greater than or equal to 10 ng/mL constitutes an adverse analytical finding (AAF). This work presents a new metabolite of higenamine in urine sample which was identified by means of ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Samples were prepared according to 2 protocols - a Dilute and Shoot (DaS) approach and a method involving acid hydrolysis and double liquid-liquid extraction (LLE). To meet the requirements typical for a confirmatory analysis, the screening procedure was further developed. In samples prepared by the DaS method, 2 peaks were observed; the earlier one was specific for higenamine and the later one unknown. MS scan analysis showed mass about 80 Da higher than that of higenamine. In turn, in samples prepared in accordance with the protocol involving hydrolysis, an increase in the area under peak for higenamine was observed, while the second peak was absent. It seems that the described strategy of detection of higenamine in urine avoids false negative results.

Section: Discussionmentioning

confidence: 99%

Analysis for higenamine in urine by means of ultra‐high‐performance liquid chromatography–tandem mass spectrometry: Interpretation of results

Grucza

Kwiatkowska

Kowalczyk

et al. 2017

“…11,12 USG has been extensively used as an alternative renal dilution factor to creatinine in toxicological applications, as well as in the field of doping control. 13 Indeed, in doping laboratories, this value is used to adjust concentrations of endogenous and exogenous substances in urine samples, [14][15][16][17][18][19][20] and critically, this value is mandatory by the World Anti-Doping Agency (WADA) in its accredited laboratories. Importantly, urine density, which is a useful measure of total urine (g/L), is based not only on the quantity, but also on the molecular mass of solute particles.…”

Section: Introductionmentioning

confidence: 99%

Development of in‐house materials for the verification of specific gravity measurements: Homogeneity, stability, and proficiency studies

Velasco-Bejarano

Mercado

Camacho

et al. 2019

We developed and evaluated the properties of in‐house urine reference materials for the verification of laboratory refractometers, which are frequently used in clinical chemistry and doping testing laboratories. Urine was gathered from 26 healthy volunteers (16 male 30 ± 5 years old and 10 female 29 ± 4 years old), from which two urine batches were obtained: one with a low specific gravity (1.012± 0.003) and the other with a high specific gravity (1.027 ± 0.003). Homogeneity studies were conducted over 20 consecutive days. For short‐term stability studies, aliquots of both urine batches were stored at −20 ± 2°C; 3 ± 2°C; 20 ± 2°C; 45 ± 2°C for 0, 2, 7, 14 and 35 days, under both light and dark conditions. Similarly, another study was conducted to measure the long‐term stability of urine at −20 ± 2°C, over a 24‐month evaluation period. Our data showed that the urine was homogeneous and stable at −20 ± 2°C, 3 ± 2°C, 20 ± 2°C, and 45 ± 2°C under both light and dark conditions. In all cases, the urine was evaluated by specific gravity and no statistically significant differences (p ≤ 0.05) were recorded. Additionally, a proficiency test was conducted in collaboration with 15 ISO/IEC 17025 accredited laboratories, and z‐scores and performance factors were evaluated. These data indicate that this material could be used for the verification of refractometers.

“…[2][3][4][5][6] The black market for AAS is expanding due to a higher consumer demand. 7 The products sold on the black market are often counterfeit and adulterated. In many cases, they contain different active pharmaceutical ingredients in lower or higher doses than declared on the label.…”

Section: Introductionmentioning

confidence: 99%

Development of an ultra‐high performance liquid chromatography–tandem mass spectrometry method for the determination of anabolic steroids currently available on the black market in the Czech Republic and Slovakia

Tirčová

Bosáková

Kozlík

2018

Over the past decade, the use of anabolic androgenic steroids (AAS) by recreational bodybuilders and regular gym members has been increasing exponentially. However, there is a lack of knowledge on AAS products sold in the Czech Republic and Slovakia. The aim of this study was to assess the quantity of active substances in AAS products obtained from AAS users in the Czech Republic and Slovakia. In addition, the study also examines the current trends in counterfeit AAS products used by recreational bodybuilders and regular gym members. For this purpose, the authors developed and validated a universal ultra‐high performance liquid chromatography with tandem mass spectrometry method to determine the most popular doping substances in different pharmaceutical formulations (oil‐based injectables, tablets, and capsules). This method was successfully utilized for the analysis of 358 AAS products. Our results showed that 58.9% AAS products analyzed contained the declared active substances at the declared concentrations, 15.9% contained no active substance, 16.8% were under‐concentrated, 4.5% contained active substances different from those declared on the label, and 3.6% products were over‐concentrated. Alarmingly, the results demonstrated that over 40% of the AAS analyzed failed to meet label claims and therefore may pose serious health risks to consumers. This study also highlighted that the availability of AAS should be more rigorously restricted and their quality closely monitored in order to protect AAS users. In conclusion, the authors have developed a precise, accurate, sensitive, selective, and robust method for the routine screening of products containing anabolic androgenic steroids.