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Urinary human chorionic gonadotropin isoform concentrations in doping control samples.

Anthony W Butch1, Getachew A Woldemariam1

  • 1UCLA Olympic Analytical Laboratory, Department of Pathology & Laboratory Medicine, Geffen School of Medicine, UCLA Health System, Los Angeles, CA, USA.

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|September 6, 2016
PubMed
Summary
This summary is machine-generated.

Anti-doping labs can get false positives for human chorionic gonadotropin (hCG) using total hCG immunoassays. Intact hCG immunoassays are better but can still overestimate; LC-MS/MS is the most accurate method for detecting hCG isoforms.

Keywords:
LC-MS/MSchorionic gonadotropinimmunoassayimmunoextractionurine

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Area of Science:

  • Clinical Chemistry
  • Biochemistry
  • Analytical Chemistry

Background:

  • Anti-doping laboratories rely on immunoassays to detect human chorionic gonadotropin (hCG) in urine.
  • Immunoassays measuring total hCG can yield false positive results due to inactive hCG isoforms like free β-subunit (hCGβ) and β-subunit core fragment (hCGβcf).
  • Intact hCG immunoassays are preferred to minimize discrepancies and false positives from these isoforms.

Purpose of the Study:

  • To determine the distribution of intact hCG, hCGβ, and hCGβcf in male urine samples.
  • To compare concentrations of hCG isoforms measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with immunoassay results.
  • To evaluate the accuracy of total and intact hCG immunoassays in identifying hCG use and atypical cases.

Main Methods:

  • Urine samples were analyzed using sequential immunoextraction with antibody-conjugated magnetic beads.
  • Concentrations of intact hCG, hCGβ, and hCGβcf were quantified using unique tryptic peptides via LC-MS/MS.
  • Results were correlated with total hCG concentrations determined by immunoassay.

Main Results:

  • Negative samples with low total hCG (<3.5 pmol/L) showed minimal intact hCG, hCGβ, and hCGβcf by LC-MS/MS.
  • Athlete urine samples after hCG administration showed high intact hCG (18.8–57.6 pmol/L) with low hCGβ but elevated hCGβcf (94–243% of intact hCG).
  • Atypical samples with high total hCG (16.7–412.7 pmol/L) by immunoassay had low intact hCG (<3.8 pmol/L) and hCGβ (<6.2 pmol/L) by LC-MS/MS, with variable hCGβcf.

Conclusions:

  • Total hCG immunoassays significantly overestimate hCG concentrations and can lead to false positives.
  • Intact hCG immunoassays offer improvement but may still slightly overestimate concentrations compared to LC-MS/MS.
  • LC-MS/MS is crucial for accurately distinguishing between legitimate hCG use and atypical cases with elevated total hCG.