Abstract

Homocysteine (Hcy), increasingly being recognized as a risk factor for vascular disease (1), is found primarily in plasma in the form of homocystine and mixed disulfides, both protein-bound and unbound; total Hcy (tHcy) is the sum of all Hcy species obtained after quantitative reduction. Several methods are currently used to measure tHcy in plasma, including GC/MS, ion-exchange chromatography, and HPLC (2). In one of the most popular HPLC techniques, any homocystine and homocysteine-mixed disulfides present are reduced with tri-butyl phosphine (TBP) to Hcy, which is subsequently derivatized with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F) to produce a fluorescent product (3)(4). TBP has a highly disagreeable odor and is poorly soluble in water so that it must be dissolved in dimethylformamide for use, both of which properties hinder the routine use of TBP in a clinical laboratory. Here, we call attention to a newer phosphine reagent, tris(2-carboxylethyl) …

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