HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (34) Citing Articles via Google Scholar Google Scholar Articles by Caussé, E. Articles by Couderc, F. Search for Related Content PubMed PubMed Citation Articles by Caussé, E. Articles by Couderc, F. Related Collections General Clinical Chemistry

Plasma Homocysteine Determined by Capillary Electrophoresis with Laser-induced Fluorescence Detection

¹ Laboratoire de Biochimie/Institut National de la Santé et de la Recherche Médicale, U466, Centre Hospitalier Rangueil, 1 avenue J. Poulhes, 31403 Toulouse Cedex 04, France;
² Laboratoire de Biochemie, Centre Hospitalier St Elio, 34295 Montpellier, France;
³ ZETA Technology, Parc Technologique du Canal, 31520 Ramonville, France;
⁴ Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, 205 Route de Narbonne, 31077 Toulouse, France;
^a author for correspondence: fax 33 5 61 322953

Homocysteine (Hcy) has emerged as another risk factor for the development of coronary heart disease (1). Genetic abnormalities of the enzymes cystathionine-ß-synthase and methylene tetrahydrofolate reductase (folic acid and vitamin B₆ and B₁₂ cofactors) can cause raised plasma Hcy concentrations. Deficiencies in folic acid and vitamins B₆ and B₁₂ may also contribute to this increased concentration (1)(2)(3). Consequently, patients with coronary heart disease have been treated successfully with folic acid, vitamin B₁₂, or vitamin B₆ (1), but the benefit of such treatment in reducing morbid cardiovascular endpoints is presently unknown (3).

A widely used technique for measuring total plasma Hcy is reversed-phase HPLC with fluorescence detection after derivatization of plasma thiols (4)(5)(6). Some studies use gas chromatography–mass spectrometry techniques (7) with various derivatization protocols (8).

The increasing demand for the determination of plasma total Hcy prompted us to develop a rapid, automated method based on capillary electrophoresis (CE) and laser-induced fluorescence detection (LIF) , in which all specific thiols are detected selectively.

Blood was collected by venipuncture into evacuated tubes containing EDTA from patients with episodes of deep-vein thrombosis or vascular disease. The plasma was separated by centrifugation and was stored at -20 °C until analysis.

Plasma samples (100 µL) were treated for 30 min at room temperature with 10 µL of a novel thiol-reducing agent [35 mmol/L tris(2-carboxyethyl)phosphine; Molecular Probes], as reported recently by Gilfix et al. (9). Compared with the widely employed tributyl phosphine, this new phosphine is nonvolatile, stable, soluble in water, and without disagreeable odor (5)(6). When we used this procedure, total Hcy (mixed and symmetric disulfides, including protein-bound Hcy) . . . [Full Text of this Article]

Acknowledgments

References

The following articles in journals at HighWire Press have cited this article:

				K. Gempel, K.-D. Gerbitz, B. Casetta, and M. F. Bauer Rapid Determination of Total Homocysteine in Blood Spots by Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry, Clin. Chem., January 1, 2000; 46(1): 122 - 123. [Full Text] [PDF]