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Automated sample preparation for the detection and confirmation of hypoxia‐inducible factor stabilizers in urine
Author(s) -
De Wilde Laurie,
Roels Kris,
Deventer Koen,
Van Eenoo Peter
Publication year - 2021
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4970
Subject(s) - chemistry , chromatography , sample preparation , solid phase extraction , urine , sample (material) , mass spectrometry , biochemistry
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.