*subcutaneous, 250 μg Ovidrel® (choriogonadotropin alfa, Sereno Inc.) injections every 3–4 days for approximately 2 weeks
2.2 | Participant selection and demographics
The study participants consisted of 10 male, healthy, and recreationally active individuals, ages 18–55. The average age of the participantswas 39 ± 5 years. The average weight of the participants was 83.6± 12.5 kg. Half of the participants were instructed to take a 20 mg/day biotin supplement and the other half was instructed to take a placebo, but no other dietary advice was given. All participants were administered hCG.
2.3 | Study drug and design
This study also monitored hCG levels of individuals supplementing with 20 mg/day of biotin or a placebo. The results of this are shownin Goodrum et al. (2023). As such, half of the individuals in this studywere taking a biotin supplement; however, we do not anticipate anyeffect of biotin supplementation on steroid profiles. An overview ofthe study design has been previously reported.6 Briefly, before beginning hCG administration, each individual provided three baseline urine and two baseline blood collections spaced apart by at least a week. Immediately after the final baseline urine and blood collections, participants began subcutaneous, 250 μg Ovidrel® (choriogonadotropin alfa, Sereno Inc.) injections every 3–4 days for approximately 2 weeks. Seven total injections were administered.Urine samples were collected daily and blood samples were collected every 3–4 days throughout the administration period and for 14 days after the final injection. On days in which an hCG injection was administered, all urine and blood collections were performed before the injection. All urine and blood collections were assigned a collection window of 24 h.
2.5 | Serum steroid profile measurements
All serum steroid profile measurements were taken using the validated liquid chromatography-mass spectometry (LC–MS) methodology compliant with the World Anti-Doping Agency (WADA) ABP Operating Guidelines8 for anti-doping analysis at the WADA accredited laboratory in Salt Lake City.
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