Chromatographic Profiling of Steroid Hormone Variability Across Menstrual Cycle and Hormonal Contraceptive Use in a Female Athlete

Natural hormones like testosterone play a big role in building and strengthening muscles. However, little is known about how the normal changes in these hormones during a woman's menstrual cycle (MC), or while taking oral hormonal contraception (HC), affect athletes who plan their training around their cycles. A study conducted by researchers at Ruhr University Bochum, the Dresden University of Technology, and the University of Wuppertal (all in Germany) as well as the Institute of Doping Analysis and Sports Biochemistry Dresden (Kreischa, Germany) investigatedvariations in steroid hormone profiles under HC and during a natural MC in one recreational athlete to bridge this knowledge gap. Hormone analysis for the subject was conducted using gas chromatography coupled with mass spectrometry (GC-MS) for urine steroids and liquid chromatography coupled with mass spectrometry (LC-MS) for plasma steroids. A paper based on this study was published in Case Reports in Endocrinology.¹

The observational study covered in the article was conducted on a 25-year-old female recreational athlete (slightly less than 5’5” in height, weighing approximately 132 lbs. and participating in about six hours of training/exercise activity per week). Testosterone and androstenedione concentrations were measured in her urine and plasma every two to three days during three weeks of HC use (hormonal vaginal ring) and the one-week HC break and 11 months later during one natural MC. Throughout the MC, urinary total androgen and plasma androstenedione levels peaked around ovulation, followed by a decline in the luteal phase, suggesting phase-dependent variation in androgen precursor availability. Meanwhile, plasma testosterone remained unchanged throughout the cycle. HC use resulted in lower androgen concentrations with no notable fluctuations in plasma or urine.¹

The fact that the MC leads to a secretion of ovarian androgens in a cycle phase-dependent manner has long been known.² With more female athletes interested in adjusting their workouts based on their menstrual cycle, the need to understand how hormones change before they can start giving solid, science-backed advice on how to train is quite clear, especially since testosterone is a proven booster for both motivation and performance, helping an athlete push harder and do better when their body is in the right state.^3-6

“This single-case study,” write the authors of the paper,¹ “demonstrates intraindividual differences in urinary and plasma androgen profiles between hormonal contraceptive use and a natural MC. While it is not possible to draw causal or performance-related conclusions from the findings, they provide descriptive insight into hormonal patterns that may inform future, larger-scale investigations.”

The researchers note that, while they tracked the menstrual cycle very accurately by monitoring body temperature, there are several limitations to their work. As they only looked at one casual athlete, they cannot be sure the results apply to professional athletes who train and recover very differently.In addition, as hormone levels in blood and urine jump around based on everyday habits (such as diet, alcohol consumption, sleep patterns, and amount of exercise), while the researchers tried to test at the same time every morning, they were unable to control everything. A longer study watching multiple cycles and looking at extra hormone markers would, in their opinion, yield much stronger data.¹

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References

1. Nolte, J.; Voss, S. C.; Keiler, A. M. et al. Intraindividual Comparison of the Anabolic Steroid Hormone Profile Between Hormonal Contraceptive Use and the Phases of a Natural Menstrual Cycle in a Recreational Athlete: A Single-Case Observational Study. Case Rep Endocrinol. 2026, 2026, 9648747. DOI: 10.1155/crie/9648747
2. Krüger, T. H. C.; Leeners, B.; Tronci, E. et al. The Androgen System Across the Menstrual Cycle: Hormonal, (Epi-)genetic and Psychometric Alterations. Physiol Behav. 2023, 259, 114034. DOI: 10.1016/j.physbeh.2022.114034
3. Ekenros, L..; Von Rosen, P..Solli G. S., et al. The Perceived Influence of Menstrual Cycle and Hormonal Contraceptives on Training and Performance: Comparison Between Football, Handball, and Orienteering, Int. J. Sports Sci. Coach. 2024, 19 (4), 1560–1569, DOI: 10.1177/17479541231216403
4. Paludo, A. C.; Paravlic, A.; Dvořáková, K. et al. The Effect of Menstrual Cycle on Perceptual Responses in Athletes: A Systematic Review With Meta-Analysis. Front Psychol. 2022, 13, 926854. DOI: 10.3389/fpsyg.2022.926854
5. Ekenros, L.; von Rosen, P.; Norrbom, J. et al. Impact of Menstrual Cycle-Based Periodized Training on Aerobic Performance, a Clinical Trial Study Protocol-The IMPACT Study. Trials 2024, 25 (1), 93. DOI: 10.1186/s13063-024-07921-4
6. Cook, C. J.; Kilduff, L. P.; Crewther, B. T. Basal and Stress-Induced Salivary Testosterone Variation Across the Menstrual Cycle and Linkage to Motivation and Muscle Power. Scand J Med Sci Sports 2018, 28 (4), 1345-1353. DOI: 10.1111/sms.13041