High-performance liquid chromatographic method for measuring total plasma homocysteine levels

Isabella Fermo, Cinzia Arcelloni, Giuseppina Mazzola, Armando D'Angelo, Rita Paroni

Research output: Contribution to journalArticlepeer-review

Abstract

We have modified a high-performance liquid chromatographic (HPLC) procedure based on SBD-F (ammonium-7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate) pre-column derivatization to obtain an assay that is useful for routine clinical total plasma homocysteine (tHcy) analysis. The introduction of easily handled sodium borohydride instead of the traditional tri-n-butylphosphine in dimethylformamide as a reductant and a 14-min run-time using basic isocratic HPLC equipment are the more notable advantages. The addition of mercaptopropionylglycine as an internal standard contributed to improvements in the reproducibility of the assay, yielding within- and between-run precisions of 1.9 and 4% (C.V.), respectively. Reference values for fasting tHcy were 7.65±2.3 and 8.9±2.4 μmol/l, while post-methionine load gave tHcy levels of 19.9±5.5 and 26.8±5.5 μmol/l, for women and men, respectively (n=40). Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)31-36
Number of pages6
JournalJournal of Chromatography B: Biomedical Applications
Volume719
Issue number1-2
DOIs
Publication statusPublished - Nov 20 1998

Keywords

  • Amino acids
  • Derivatization, LC
  • Homocysteine
  • Methionine

ASJC Scopus subject areas

  • Chemistry(all)

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