Determination of free and total homocysteine in human plasma by high-performance liquid chromatography with fluorescence detection

https://doi.org/10.1016/0378-4347(87)80438-3Get rights and content

Abstract

A simple, sensitive and precise method for the determination of both free and total homocysteine in human plasma is presented. The total homocysteine in plasma consists of free homocysteine (i.e. reduced plus oxidized homocysteine in the non-protein fraction of plasma) and protein-bound homocysteine. The thiol compounds in plasma, which are reduced or liberated from plasma proteins with tri-n-butylphosphine, are derivatized with a thiol-specific fluorogenic reagent, ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate. The derivatives are separated by reversed-phase high-performance liquid chromatography. The concentrations (mean ±S.D.) of free and total homocysteine in plasma from 35 normal subjects were 1.94±0.46 and 6.18±1.19 nmol/ml, respectively.

References (27)

  • R.T. Wall et al.

    Thromb. Res.

    (1980)
  • R. Saetre et al.

    Anal. Biochem.

    (1978)
  • G.L. Newton et al.

    Anal. Biochem.

    (1981)
  • K. Imai et al.

    Anal. Biochem.

    (1983)
  • T. Toyo'oka et al.

    J. Chromatogr.

    (1983)
  • T. Toyo'oka et al.

    J. Pharm. Biomed. Anal.

    (1984)
  • L.A. Smolin et al.

    J. Nutr.

    (1982)
  • A.M. Bond et al.

    Anal. Chim. Acta

    (1984)
  • S.B. Thomson et al.

    J. Chromatogr.

    (1985)
  • A. Svardal et al.

    J. Biol. Chem.

    (1986)
  • G.W. Barber et al.

    J. Pediatr.

    (1969)
  • A.J.L. Cooper

    Ann. Rev. Biochem.

    (1983)
  • K.S. McCully

    Am. J. Pathol.

    (1969)
  • Cited by (1016)

    • Plasma Homocysteine and Glutathione Are Independently Associated With Estimated Glomerular Filtration Rates in Patients With Renal Transplants

      2019, Transplantation Proceedings
      Citation Excerpt :

      Plasma malondialdehyde (MDA) concentration was determined with thiobarbituric acid-reactive substances to indicate the level of oxidative stress [9]. Plasma PLP [10], homocysteine, and cysteine [11] were quantified by high-performance liquid chromatography using a fluorescence detector. Hyperhomocysteinemia was defined as plasma homocysteine concentrations ≥ 14 mmol/L [12].

    View all citing articles on Scopus
    View full text