Metabolism of boldione in humans by mass spectrometric techniques: detection of pseudoendogenous metabolites

Drug Test Anal. Nov-Dec 2013;5(11-12):834-42. doi: 10.1002/dta.1567. Epub 2013 Nov 20.


Boldione is an anabolic androgenic steroid (AAS) related to boldenone, androstenedione, and testosterone bearing two double bonds in C1 and C4 positions. Boldione is rapidly transformed to the well-known AAS boldenone, being both compounds included in the list of prohibited substances and methods published yearly by the World Anti-Doping Agency (WADA). After the administration of boldione to a male volunteer, the already described urinary metabolites of boldenone produced after reduction in C4, oxydoreduction in C3 and C17, and hydroxylation have been detected. In addition, minor new metabolites have been detected and their structure postulated after mass spectrometric analyses. Finally, the reduction of the double bound in C1 produces metabolites identical to the endogenously produced ones. A method based on gas chromatography coupled to isotope ratio mass spectrometry (GC/C/IRMS) after a urine sample purification by high performance liquid chromatography (HPLC) permitted to confirm the main synthetic like boldione/boldenone metabolite (17β-hydroxy-5β-androst-1-en-3-one) and boldenone at trace levels (< 5 ng/mL) and then to establish its synthetic or endogenous origin, and to determine the exogenous origin of metabolites with the same chemical structure of the endogenous ones. The detection of pseudoendogenous androgens of synthetic origin partially overlapped boldenone and its main metabolite detection, being an additional proof of synthetic steroids misuse. By the use of IRMS, the correct evaluation of the modifications of the steroid profile after the administration of synthetic AAS that could be converted into endogenous like ones is possible.

Keywords: Boldione; IRMS; LC sample treatment; MS; anabolic steroids; androgens; anti-doping.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adult
  • Anabolic Agents / metabolism*
  • Anabolic Agents / urine*
  • Chromatography, High Pressure Liquid / methods
  • Gas Chromatography-Mass Spectrometry / methods*
  • Humans
  • Male
  • Mass Spectrometry / methods
  • Sensitivity and Specificity
  • Substance Abuse Detection / methods
  • Testosterone / analogs & derivatives*
  • Testosterone / metabolism
  • Testosterone / urine


  • Anabolic Agents
  • Testosterone
  • boldenone