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Automated sample preparation for the detection and confirmation of hypoxia-inducible factor stabilizers in urine.

Authors
  • De Wilde, Laurie1
  • Roels, Kris1
  • Deventer, Koen1
  • Van Eenoo, Peter1
  • 1 Doping Control Laboratory, Ghent University, Department Diagnostic Sciences, Zwijnaarde, Belgium. , (Belgium)
Type
Published Article
Journal
Biomedical Chromatography
Publisher
Wiley (John Wiley & Sons)
Publication Date
Feb 01, 2021
Volume
35
Issue
2
Identifiers
DOI: 10.1002/bmc.4970
PMID: 32840903
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

As hypoxia-inducible factor stabilizers (HIFs) can artificially enhance an athlete's erythropoiesis, the World Anti-Doping Agency prohibits their use at all times. Every urine sample for doping control analysis has to be evaluated for the presence of HIFs and therefore sensitive methods that allow high sample throughput are needed. Samples suspicious for the presence of HIFs need to be confirmed following the identification criteria established by the World Anti-Doping Agency. Previous work has shown the advantages of using turbulent flow online solid-phase extraction (SPE) procedures to reduce matrix effects and retention time shifts. Furthermore, the use of online SPE allows for automation and high sample throughput. Both an initial testing procedure (ITP) and a confirmation method were developed and validated, using online SPE liquid chromatography-tandem mass spectrometry (LC-MS/MS), with limits of detection between 0.1 ng/ml (or possibly lower) and 4 ng/ml (or higher for GSK360a) and limits of identification between 0.1 ng/ml (or possibly lower) and 1.17 ng/ml. The ITP only takes 6.5 min per sample. To the best of our knowledge, these are the first ITP and confirmation methods that include more than three HIFs without the need for manual sample preparation. © 2020 John Wiley & Sons, Ltd.

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