Determination of (13)C/(12)C ratios of urinary excreted boldenone and its main metabolite 5beta-androst-1-en-17beta-ol-3-one.

The research article discusses the use of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) to analyze the carbon isotope ratios in the anabolic steroid Boldenone and its main metabolite, with the aim of discerning whether the substances originated from within the body or were introduced externally. The findings suggest that some athletes may naturally produce the banned substance.

Background and Aim of the Research

• The researchers aim to tackle an ongoing controversy in the field of sports science and doping control. Boldenone, an anabolic steroid not clinically approved for humans, has been detected in several doping control samples. The significant question at hand is whether Boldenone can be endogenously produced (i.e. produced naturally within the human body) or if its detection is purely indicative of exogenous doping (introducing the substance from outside the body).
• The team proposes a method by which the origin of Boldenone (and its main metabolite, BM1) can be traced accurately. The method involves measuring the ratio of two carbon isotopes, (13)C and (12)C, using GC/C/IRMS.

Method Development

• The researchers developed a method to determine the (13)C/(12)C ratios of Boldenone, BM1, and a few other substances from a single urine sample. This method is essentially built to distinguish the endogenous or exogenous source of these substances.
• To ensure the validity of the method, repeated processing of urine fortified with various concentrations of Boldenone and BM1 was performed. Specificity of the method was ensured through gas chromatography/mass spectrometry determinations.

Findings from the Research

• The newly developed method was applied to 23 samples collected over a four-year period. Findings from the isotope analysis suggested that in 11 out of these 23 samples, the Boldenone or its metabolite BM1 had carbon isotope ratios inconsistent with an exogenous origin.
• Two of these samples were collected from the same athlete within a one-month interval, strongly indicating the possibility of endogenous Boldenone production in this particular athlete.

Implications of the Findings

• The findings of this study have significant implications for doping control in sports. If Boldenone or its metabolite BM1 can be endogenously produced, even in few individuals, it would mean the current methods of doping detection could generate false positives.
• The developed method could be used by doping control agencies to accurately determine the source of the detected Boldenone or BM1, hence ensuring fair and accurate doping prosecution.