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

Laurie De Wilde (UGent) , Kris Roels (UGent) , Koen Deventer (UGent) and Peter Van Eenoo (UGent)
(2021) BIOMEDICAL CHROMATOGRAPHY. 35(2).
Author
Organization
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.
Keywords
Analytical Chemistry, Clinical Biochemistry, Biochemistry, Molecular Biology, Pharmacology, Drug Discovery, General Medicine, doping, hypoxia-inducible factor stabilizers, LC-MS, MS, turbulent flow online SPE, urine, DILUTE-AND-SHOOT, CHROMATOGRAPHY-MASS-SPECTROMETRY, TURBULENT-FLOW CHROMATOGRAPHY, SOLID-PHASE EXTRACTION, DOPING CONTROL, ONLINE CLEANUP, LC-MS/MS, AGENTS, METABOLITES, STIMULANTS

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Citation

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MLA
De Wilde, Laurie, et al. “Automated Sample Preparation for the Detection and Confirmation of Hypoxia‐inducible Factor Stabilizers in Urine.” BIOMEDICAL CHROMATOGRAPHY, vol. 35, no. 2, 2021, doi:10.1002/bmc.4970.
APA
De Wilde, L., Roels, K., Deventer, K., & Van Eenoo, P. (2021). Automated sample preparation for the detection and confirmation of hypoxia‐inducible factor stabilizers in urine. BIOMEDICAL CHROMATOGRAPHY, 35(2). https://doi.org/10.1002/bmc.4970
Chicago author-date
De Wilde, Laurie, Kris Roels, Koen Deventer, and Peter Van Eenoo. 2021. “Automated Sample Preparation for the Detection and Confirmation of Hypoxia‐inducible Factor Stabilizers in Urine.” BIOMEDICAL CHROMATOGRAPHY 35 (2). https://doi.org/10.1002/bmc.4970.
Chicago author-date (all authors)
De Wilde, Laurie, Kris Roels, Koen Deventer, and Peter Van Eenoo. 2021. “Automated Sample Preparation for the Detection and Confirmation of Hypoxia‐inducible Factor Stabilizers in Urine.” BIOMEDICAL CHROMATOGRAPHY 35 (2). doi:10.1002/bmc.4970.
Vancouver
1.
De Wilde L, Roels K, Deventer K, Van Eenoo P. Automated sample preparation for the detection and confirmation of hypoxia‐inducible factor stabilizers in urine. BIOMEDICAL CHROMATOGRAPHY. 2021;35(2).
IEEE
[1]
L. De Wilde, K. Roels, K. Deventer, and P. Van Eenoo, “Automated sample preparation for the detection and confirmation of hypoxia‐inducible factor stabilizers in urine,” BIOMEDICAL CHROMATOGRAPHY, vol. 35, no. 2, 2021.
@article{8673888,
  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.}},
  articleno    = {{e4970}},
  author       = {{De Wilde, Laurie and Roels, Kris and Deventer, Koen and Van Eenoo, Peter}},
  issn         = {{0269-3879}},
  journal      = {{BIOMEDICAL CHROMATOGRAPHY}},
  keywords     = {{Analytical Chemistry,Clinical Biochemistry,Biochemistry,Molecular Biology,Pharmacology,Drug Discovery,General Medicine,doping,hypoxia-inducible factor stabilizers,LC-MS,MS,turbulent flow online SPE,urine,DILUTE-AND-SHOOT,CHROMATOGRAPHY-MASS-SPECTROMETRY,TURBULENT-FLOW CHROMATOGRAPHY,SOLID-PHASE EXTRACTION,DOPING CONTROL,ONLINE CLEANUP,LC-MS/MS,AGENTS,METABOLITES,STIMULANTS}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{10}},
  title        = {{Automated sample preparation for the detection and confirmation of hypoxia‐inducible factor stabilizers in urine}},
  url          = {{http://dx.doi.org/10.1002/bmc.4970}},
  volume       = {{35}},
  year         = {{2021}},
}
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