Fluorescence enhancement of PLP for application to HPLC.

Abstract

In an HPLC analysis of vitamin B6 derivatives, the use of a reversed-phase ODS column in conjunction with a fluorescence detector has given promising results. However, PLP was rather insensitive to the analysis, making it difficult to obtain accurate results, especially in biological materials. Subsequently, a pre-column treatment that involved extracting the sample solution with 1 M perchloric acid, heating the extract for 3 hr at 50°C in the presence of 5mM KCN at pH 7.5, and then allowing it to stand for 24hr at room temperature served to overcome difficulties in the analysis. The fluorescence enhancement of PLP was over 10-fold and produced a single peak that was applicable to the quantification of PLP in biological materials.