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

J Chromatogr B Biomed Sci Appl. 1998 Nov 20;719(1-2):31-6. doi: 10.1016/s0378-4347(98)00405-8.


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 micromol/l, while post-methionine load gave tHcy levels of 19.9+/-5.5 and 26.8+/-5.5 micromol/l, for women and men, respectively (n=40).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Female
  • Fluorescent Dyes
  • Fluorobenzenes
  • Homocysteine / blood*
  • Humans
  • Male
  • Reference Values
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence


  • Fluorescent Dyes
  • Fluorobenzenes
  • Homocysteine
  • 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate