Journal
CLINICA CHIMICA ACTA
Volume 331, Issue 1-2, Pages 19-23Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0009-8981(03)00076-7
Keywords
homocysteine; serum; plasma; HPLC; reversed phase
Categories
Ask authors/readers for more resources
Background: Determination of plasma homocysteine has gained increasing interest during the past few years. Several HPLC methods for determination of homocysteine are available. Based on these methods, we developed a new HPLC assay for rapid and sensitive measurement of total plasma homocysteine. Methods: As a reducing reagent tris-(2-carboxylethyl)-phosphine is used, ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate serves as the derivatization agent. Separation is performed by reversed-phase HPLC using a precolumn and a 55-mm RP(18) cartridge; mobile phase: 0.1 mol/l KH(2)PO(4) with 5% methanol, adjusted to pH 2.7 with ortho-phosphoric acid, flow-rate 1.1 ml/min. Results: Homocysteine is clearly separated from other thiols, the retention time being 2.2 min, total analysis time is 6 min. Tests for linearity, recovery and precision are satisfactory, as well as the comparison with a commercial available assay method. Detection limit of the method is 0.5 mumol/l, it could be further enhanced for measurements of even lower homocysteine concentrations in, e.g., cell culture supernatants. Conclusions: The described method is well suited for analysis of thiols in blood specimens. It is more convenient and more rapid than methods described earlier. (C) 2003 Elsevier Science B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available