Improving the Determination of Carbon Isotope Ratios of Endogenous Steroids Found in Human Serum
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View abstract on PubMed
Summary
This summary is machine-generated.This study introduces a validated method for analyzing serum testosterone and 4-androstenedione isotope ratios, aiding in the detection of doping in athletes. The method enhances the Athlete Biological Passport by identifying exogenous steroid use.
Area Of Science
- Sports Science
- Analytical Chemistry
- Biochemistry
Background
- The Athlete Biological Passport (ABP) now includes serum testosterone (T) and 4-androstenedione (A4) to detect low-dose T misuse, particularly in female athletes.
- Unlike urinary steroid profiles, confounding factors for serum T and A4 are less understood, necessitating methods to confirm exogenous sources via isotope ratio mass spectrometry (IRMS).
Purpose Of The Study
- To develop and validate a comprehensive method for determining carbon isotope ratios (CIRs) in serum steroids.
- To establish robust protocols for doping control laboratories to identify exogenous steroid administration.
Main Methods
- A high-performance liquid chromatography (HPLC) cleanup method was developed and validated for serum steroids.
- Two-dimensional gas chromatography coupled with isotope ratio mass spectrometry (2D-GC/IRMS) was employed for CIR determination.
- Analyte purification strategies were enhanced to include cholesterol, dehydroepiandrosterone sulfate, androsterone sulfate, epiandrosterone sulfate, pregnenolone sulfate, and 5-androstene-3β,17β-diol sulfate.
Main Results
- Robust CIR determinations were achieved for serum steroid concentrations as low as 25 ng/mL.
- A reference population of 124 male and female athletes was analyzed.
- Population-based thresholds for steroid combinations were calculated, supporting the interpretation of CIR data.
Conclusions
- The validated HPLC-2D-GC/IRMS method provides a reliable tool for doping control laboratories to assess serum steroid CIRs.
- This approach enhances the detection capabilities of the ABP for exogenous steroid administration, complementing existing urinary analyses.
- The establishment of population-based thresholds aids in the accurate interpretation of longitudinal steroid profiles.
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