Combination of liquid-chromatography tandem mass spectrometry in different scan modes with human and chimeric mouse urine for the study of steroid metabolism

Oscar J Pozo, Leen Lootens, Peter Van Eenoo, Koen Deventer, Philip Meuleman, Geert Leroux-Roels, Maria K Parr, Wilhelm Schänzer, Frans T Delbeke

Publication: Contribution to journalJournal articlesResearch

Abstract

Anabolic steroids are among the most frequently detected compounds in doping analysis. They are extensively metabolized and therefore an in-depth knowledge about steroid metabolism is needed. In this study, a liquid chromatography tandem mass spectometry (LC-MS/MS) method based on a precursor ion scan with a uPA-SCID mouse with humanized liver (a chimeric mouse) was explored for the detection of steroid metabolism. Methandienone was used as a model compound. The application of the precursor ion scan method in positive human samples and chimeric mice samples after methandienone administration allowed the detection of most steroid metabolites without any structural restriction. Three hitherto unreported metabolites were found using this approach. These metabolites were characterized using LC-MS/MS and feasible structures were proposed. The structure of one of them, 6-ene-epimethandienone, was confirmed by the synthesis of the reference compound. A selected reaction monitoring (SRM) method for the specific detection of all these metabolites has been developed. The application of this method to several human and chimeric mouse samples confirmed that more than 80% of the steroid metabolites were found in both samples. Only metabolites that are poorly detectable by LC-MS/MS were not detected in some urine samples. The metabolic nature of the unreported metabolites was also confirmed. A global strategy for the detection of steroid metabolites combining both human and chimeric mouse urine is proposed.

Original languageEnglish
JournalDrug testing and analysis
Volume1
Issue number11-12
Pages (from-to)554-567
Number of pages14
DOIs
Publication statusPublished - 01.11.2009

Research areas and keywords

  • Anabolic Agents
  • Animals
  • Chromatography, Liquid
  • Doping in Sports
  • Hepatocytes
  • Humans
  • Methandrostenolone
  • Mice
  • Mice, SCID
  • Molecular Structure
  • Specimen Handling
  • Substance Abuse Detection
  • Tandem Mass Spectrometry
  • Transplantation Chimera

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