Plasma total homocysteine quantification: an improvement of the classical high-performance liquid chromatographic method with fluorescence detection of the thiol-SBD derivatives

A rapid, shorter, isocratic high-performance liquid chromatographic method is described for the determination of plasma total homocysteine. In this method the sample preparation was modified for reduction of the time of the thiolic reduction from 30 at room temperature to 10 min at 37 °C with tris-(2-carboxyethyl) phosphine (TCEP), reduction the time of derivatization from 60 to 10 min at 60 °C and elution of the SBD-thiols derivative by a shorter HPLC-column which is commercially available. The SBD–homocysteine derivative was eluted at 3.7 min. The method was equally precise and faster for quantification of tHcy in plasma as other previously described method and should be very useful for epidemiological studies in which large numbers of samples have to be analyzed