An on-column derivatization method for the determination of homocysteine-thiolactone and protein N-linked homocysteine

Rafał Głowacki, Edward Bald, Hieronim Jakubowski

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

Homocysteine (Hcy) is incorporated into protein via a reaction of the thioester Hcy-thiolactone with ε-amino group of a protein lysine residue generating N-Hcy-protein. This reaction impairs and alters protein's function and has been implicated in atherothrombotic disease. Here, we describe new high-performance liquid chromatography assays for the determination of Hcy-thiolactone, protein N-linked Hcy, and Hcy based on an on-column derivatization with o-phthaldialdehyde and fluorescence detection. The on-column derivatization generates narrow peaks, which allows fast run times (3-5 min) and facilitates determination of N-linked Hcy directly from acid hydrolysates of plasma protein. Utility of these assays was demonstrated with human urine and plasma samples.

Original languageEnglish (US)
Pages (from-to)187-194
Number of pages8
JournalAmino Acids
Volume41
Issue number1
DOIs
StatePublished - Jun 1 2011

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Clinical Biochemistry
  • Organic Chemistry

Keywords

  • HPLC
  • Homocysteine-thiolactone
  • Human plasma
  • O-Phthaldialdehyde
  • Protein N-linked homocysteine
  • Urine

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