Simultaneous Determination of Glycated Albumin and D-Glucose in Human Serum by High-Performance Liquid Chromatography with Postcolumn Fluorescence Derivatization

High-performance liquid chromatography with postcolumn fluorescence derivatization for the simultaneous determination of glycated albumin and D-glucose in human serum is described. Glycated albumin and D-glucose were separated along with other glycated proteins on a size exclusion column (TSKgel G2000SWXL) and converted into fluorescence derivatives by reaction with 4-methoxybenzamidine or meso-1, 2-bis(4-methoxyphenyl)ethylenediamine in an alkaline medium. The method required only 1μl of serum. The detection limits for glycated albumin and D-glucose (pmol on column) were 100 and 3 with the former reagent, and 150 and 5 with the latter, respectively.