Immature and mature collagen crosslinks quantification using high-performance liquid chromatography and high-resolution mass spectrometry in orbitrap™

Diana R. Hernandez, Maria del Carmen Piqueras, Alejandro E. Macias, Laisel Martinez, Roberto Vazquez-Padron, Sanjoy K. Bhattacharya

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Different methodologies for collagen quantification have been described in the past. Introduction of mass spectrometry combined with high-performance liquid chromatography (HPLC) is a high-resolution tool, which has generated novel applications in biomedical research. In this study, HPLC coupled to electrospray ionization (ESI) tandem mass spectrometry (HPLC-ESI-MS/MS) was used to characterize tissue samples from AVFs done in rats. These findings helped create a protocol for identifying and quantifying components of immature and mature collagen crosslink moieties. Two different internal standards were used: epinephrine and pyridoxine. Quantification curves were drawn by means of these standards. The goal of the experiment was to achieve accurate quantification with the minimum amount of sample. Time and cost of experiment were considerably minimized. Up to date, this method has not been tested for crosslinking quantification.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages101-111
Number of pages11
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1996
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Amino acids
  • Arteriovenous fistula
  • Collagen
  • Crosslinking
  • Orbitrap
  • Q exactive

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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