lmplementation of the prolyl hydroxylase inhibitor Roxadustat (FG-4592) and its main metabolites into routine doping controls

lmplementation of the prolyl hydroxylase inhibitor Roxadustat (FG-4592) and its main metabolites into routine doping controls /  Daniel Eichner, Ryan M. Van Wagoner, Mitch Brenner, James Chou, Scott Leigh, Lee R. Wright, Lee A. Flippin, Michael Martinelli, Oliver Krug, Wilhelm Schänzer, Mario Thevis

  • Drug Testing and Analysis 9 (2017) 11-12 (November-December), p. 1768-1778)
  • Special Issue: 35th Cologne workshop: Advances in sports drug testing
  • PMID: 28378453
  • DOI: 10.1002/dta.2202


The utility of hypoxia-inducible factor (HIF) prolyl hydroxylase inhibitors as a therapeutic means of treating patients suffering from anaemia has been demonstrated for various clinical settings. However, besides this intended use, HIF stabilizers can be the subject of misuse in amateur and elite sports due to their erythropoietic properties, as recently proven by several cases of adverse analytical findings in doping control testing. Consequently, to allow for adequate and comprehensive test methods, knowledge of the drug candidates' metabolism and analytical options enabling appropriate detection windows in sports drug testing samples (i.e., blood and urine) is essential to doping control laboratories. In the present study, a novel HIF prolyl hydroxylase inhibitor referred to as Roxadustat (FG-4592) and main plasma- and urine-derived metabolites were investigated in the context of routine doping control analytical approaches. Liquid chromatography-mass spectrometry-based test methods were used to study the target analytes' dissociation pathways following electrospray ionization and collision-induced dissociation. Diagnostic precursor-product ion pairs were selected to enable the implementation of the intact drug Roxadustat and selected metabolites into multi-analyte initial testing procedures for plasma and urine specimens. The assays were validated in accordance to guidelines of the World Anti-Doping Agency (WADA) and results demonstrated the suitability (fitness-for-purpose) of the employed analytical methods with detection limits ranging from 0.05 to 1 ng/mL and 1 to 5 ng/mL for urine and plasma, respectively. Subsequently, elimination study plasma and urine samples collected up to 167 h post-administration were analyzed using the validated methods, which suggested the use of different target analytes for blood and urine analyses with FG-4592 and its glucuronide, respectively, for optimal detection windows. Additionally, a light-induced rearrangement product (photoisomer) of Roxadustat resulted in the formation of an additional compound of identical mass.

Original document


Research / Study
31 May 2017
Brenner, Mitch
Chou, James
Eichner, Daniel
Flippin, Lee A.
Krug, Oliver
Leigh, Scott
Martinelli, Michael
Schänzer, Wilhelm
Thevis, Mario
Van Wagoner, Ryan M.
Wright, Lee R.
United States of America
Other organisations
Deutsche Sporthochschule Köln (DSHS) - German Sport University Cologne
European Monitoring Center for Emerging Doping Agents (EuMoCEDA)
Salt Lake City, USA: The Sports Medicine Research and Testing Laboratory (SMRTL)
Analytical aspects
Mass spectrometry analysis
Testing method development
Doping classes
S2. Peptide Hormones, Growth Factors
Hypoxia-inducible factor (HIF) activating agents
Roxadustat (FG-4592)
Document category
Scientific article
Document type
Pdf file
Date generated
1 November 2022
Date of last modification
16 November 2022
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