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Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is...
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QuEChERS as alternative extraction procedure in doping analyses.

Daniel Derwand1,2, Sebastian Rzeppa1, Sven C Voss1

  • 1Institute of Doping Analysis and Sports Biochemistry Dresden, Kreischa, Germany.

Drug Testing and Analysis
|December 3, 2023
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Summary
This summary is machine-generated.

A new QuEChERS method for urine sample preparation in doping control efficiently extracts 312 compounds. This cost-effective technique, using ammonium acetate and formic acid, meets World Anti-Doping Agency requirements for detecting prohibited substances.

Keywords:
HIF activating agentdispersive solid phase extractionstanozolol‐glucuronides

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Area of Science:

  • Analytical Chemistry
  • Forensic Science
  • Biochemistry

Background:

  • QuEChERS (Quick, Easy, Cheap, Rugged, and Safe) is a widely used sample preparation technique in food analysis for detecting residues like pesticides and mycotoxins.
  • Traditional methods for doping control sample preparation can be time-consuming and costly.
  • The World Anti-Doping Agency (WADA) sets stringent performance requirements for analytical methods used in doping control.

Purpose of the Study:

  • To evaluate a QuEChERS-based method for preparing urine samples for doping control.
  • To assess the efficiency of a specific QuEChERS protocol (ammonium acetate/formic acid in acetonitrile) for extracting a broad range of doping agents.
  • To validate the method according to WADA requirements for confirmation procedures.

Main Methods:

  • A QuEChERS method utilizing 10 M ammonium acetate and 3% formic acid in acetonitrile was developed for urine sample preparation.
  • The method was tested on urine samples with varying pH and specific gravity.
  • Extraction efficiency was assessed for 342 compounds, with a focus on nine selected analytes representing six WADA Prohibited List categories.

Main Results:

  • The optimized QuEChERS method successfully extracted 312 out of 342 tested compounds at their minimum required performance levels.
  • The method demonstrated high efficiency for extracting nine selected analytes, meeting WADA's criteria for confirmation procedures.
  • Specific attention was given to the extraction of intact stanozolol-glucuronides, showing promising results for high-resolution mass spectrometry analysis.

Conclusions:

  • The developed QuEChERS method offers a time- and cost-effective alternative for urine sample preparation in doping control.
  • This approach can be a valuable tool for the confirmation analysis of non-threshold prohibited substances.
  • The method's efficiency and compliance with WADA standards make it a viable option for anti-doping laboratories.