Quantitative method for biomarkers of collagen degradation using liquid chromatography tandem mass spectrometry

Anal Biochem. 2000 Jul 15;283(1):71-6. doi: 10.1006/abio.2000.4625.

Abstract

Preclinical efficacy testing commonly involves studies that require considerable resources and time. One valuable tool in this endeavor is the characterization of relevant biomarkers. A method has been developed for the simultaneous determination of collagen biomarker candidates as an instrument in screening compounds for efficacy. Two potential candidates, the 3-hydroxypyridinium crosslinks pyridinoline and deoxypyridinoline, were selected for analysis in collagen degradation models. Tissue or urine samples were collected, prepared, and quantitated for the biomarkers using spiked calibration curves and liquid chromatography tandem mass spectrometry. The development of a quick and simple assay method would allow us to increase the chances for success in efficacy screening by eliminating compounds with poor biomarker profiles. The method proposed here appears to be more selective, convenient, precise (generally <10% RSD), accurate (generally <10% RE), and sensitive relative to previously established methodology.

MeSH terms

  • Amino Acids / chemistry
  • Amino Acids / metabolism
  • Amino Acids / urine
  • Animals
  • Biomarkers*
  • Collagen / analysis*
  • Collagen / metabolism*
  • Collagen / urine
  • Gas Chromatography-Mass Spectrometry / methods*
  • Male
  • Matrix Metalloproteinase Inhibitors
  • Pyridines / chemistry
  • Rabbits
  • Rats
  • Rats, Wistar
  • Time Factors

Substances

  • Amino Acids
  • Biomarkers
  • Matrix Metalloproteinase Inhibitors
  • Pyridines
  • 3-hydroxypyridine
  • pyridinoline
  • deoxypyridinoline
  • Collagen