Growth hormone abuse in the horse: preliminary assessment of a mass spectrometric procedure for IGF-1 identification and quantitation

Rapid Commun Mass Spectrom. 2001;15(14):1191-7. doi: 10.1002/rcm.363.

Abstract

Previous studies have shown that insulin-like growth factor 1 (IGF-1) is a promising marker for the detection of growth hormone (GH) abuse in the horse. The significant increases observed with GH administration in comparison to natural levels imply the possibility of setting a threshold level for IGF-1 that would be indicative of GH abuse. Although an immunoradiometric assay (IRMA) has been identified as a reliable screening method, a more specific IGF-1 quantification method needs to be developed for the prosecution of GH abuse by horseracing authorities. This study describes such an HPLC electrospray mass spectrometry (LC/ESI-MS) method that was developed and then assessed for the specific analysis of IGF-1 at the low levels encountered in serum. The structural identity of IGF-1 was confirmed by endoproteinase Asp-N digestion followed by LC/MS and LC/MS/MS characterisation. This was followed by quantification of IGF-1 as the intact molecule against an internal standard.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biomarkers / analysis
  • Chromatography, High Pressure Liquid
  • Endopeptidases / metabolism
  • Growth Hormone / blood*
  • Growth Hormone / pharmacology
  • Horses*
  • Insulin-Like Growth Factor I / analysis
  • Insulin-Like Growth Factor I / chemistry*
  • Metalloendopeptidases
  • Molecular Sequence Data
  • Recombinant Proteins / analysis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Serine Endopeptidases / metabolism
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Substance Abuse Detection / methods*

Substances

  • Biomarkers
  • Recombinant Proteins
  • Insulin-Like Growth Factor I
  • Growth Hormone
  • Endopeptidases
  • Serine Endopeptidases
  • glutamyl endopeptidase
  • Metalloendopeptidases
  • endoproteinase Asp-N