madman
Super Moderator
Abstract
The Athlete Biological Passport (ABP) is a longitudinal tool used in anti-doping to monitor biological parameters known to change with performance-enhancing drug use. The ABP consists of multiple modules, including two aimed at detecting the use of endogenous anabolic androgenic steroids: the urinary and serum steroid modules. Human chorionic gonadotropin (hCG) is a protein hormone potentially abused by male athletes to increase the production of endogenous testosterone. To date, no studies have investigated the impact of extended hCG administration on the urinary and serum steroid modules of the ABP. The goal of this study was to identify the impact of multiple hCG administrations on the parameters tracked as part of the urinary and serum steroid modules of the ABP. Ten recreationally active, healthy male individuals self-administered seven 250 μg hCG injections over 3 weeks. Serum and urine samples were collected before, during, and 2 weeks following the final injection. All ABP parameters were quantified in the respective matrix, and steroid profiles were created with Anti-Doping Administration and Management System adaptive model upper and lower limits for both matrices. In both serum and urine profiles, testosterone increased; however, the testosterone/epitestosterone ratio in urine and the testosterone/androstenedione ratio in serum showed minimal changes. Additionally, serum luteinizing hormone (LH) was quantified using an immunoassay, and a serum testosterone/LH ratio was generated. Serum LH values decreased during administration causing large increases in the serum T/LH ratio, indicating this ratio may be a more sensitive parameter for detecting hCG abuse than urinary testosterone/epitestosterone or serum testosterone/androstenedione.
3.2 | Median serum steroid profile changes
The median percent change and interquartile ranges (IQR) observed among all participants for the three serum steroidal module parameters are shown in Figure 1. After administration of hCG, T increased over 100% and remained elevated throughout the administration period before decreasing approximately 5 days after the final hCG dose and falling below baseline at the final collection, about 2 weeks after the final dose (Figure 1a). A4 follows a similar trend to T with identical timing and a similar magnitude of increase and decrease(Figure 1b); however, the T/A4 ratio remained stable with relatively minimal changes (Figure 1c).
3.3 | Individual serum steroid profile changes
To put these changes into the context of the ABP, individual profiles were generated for all participants with upper and lower limits generated by the ADAMS adaptive model. Three representative individuals are shown in Figure 2. Nine out of 10 participants had at least one T value that exceeded the upper limit during the administration period, as seen in HCG1, HCG5, and HCG10. Five out of 10 participants had at least one T value below the lower limit during the washout phase, as seen in HCG1 and HCG5. For the T/A4 ratio, only two individuals had a single value below the lower limit, observed during the washout phase, and two participants had at least one value above the upper limit (HCG10; Figure 2c) observed during the administration period.
3.6 | Changes in serum LH and the serumT/LH ratio
To investigate serum LH as a potential biomarker of endogenous anabolic androgenic steroids and/or hCG abuse, serum LH values were quantified using the Roche Cobas e411 immunoanalyzer, and the median percent change and IQR for all participants can be found in Figure 5a. LH values significantly decreased after the first hCG injection and remained suppressed throughout the entire administration period before beginning to recover approximately 2 weeks after the final hCG administration. T values from the serum steroidal module were used to create a T/LH ratio. The median percent changes with IQR of the T/LH ratio for all participants are shown in Figure 5b. This ratio becomes elevated after the first hCG injection before showing a substantial increase in all participants through the administration period and recovering to baseline at the final collection 2 weeks after the final dose. Individual profiles of the serum T/LH ratio for three representative individuals are shown in Figure 5c.
One notable limitation of the study design is the length of timewash out samples were collected. The final collection in both urine and serum was performed 11–14 days after the last hCG injection was administered and it is clear that a number of parameters did not return to baseline in this time frame, despite hCG levels falling below the positivity criteria by the final collection.6 These parameters include serum T, A4, and the T/A4 ratio (Figure 1), serum LH(Figure 5a), and the urinary T/E, A/T, and 5α-Adiol/E ratios(Figure 3). In fact, it appears some of the most dramatic changes appear to have happened in the last few collections (Figure 3b,e),and as such, the timing of the return to baseline for these parameters would have been useful to observe. An additional limitation is the presence of high levels of biotin in the odd-numbered participants urine. While we do not anticipate any effects on the steroid profiles of these individuals, these biotin levels prohibited accurate immunoassay testing for urinary LH and intact hCG using the RocheCobas immunoanalyzer. However, total urinary hCG levels measured with a biotin-independent assay can be seen in Goodrum et al.(2023).6
5 | CONCLUSIONS
Extended hCG administration induces major changes in serum T and A4 levels, with minor changes to the T/A4 ratio tracked in the serum steroid module of the ABP. For the urinary steroid module, the T/E ratio showed a modest increase during the administration period and the 5α-Adiol/E and A/T ratios showed major changes during the washout period, while the 5α-Adiol/5β-Adiol and A/Etio ratios did not show any changes. Major changes in the serum T/LH ratio were observed throughout the administration period and up to 9 days after, indicating this parameter may be a worthwhile additionto the serum steroidal module in the future as a sensitive biomarkerfor hCG use.
