High-performance liquid chromatographic assay of serum glycated albumin, with special reference to a comparison of its assay conditions with those of fructosamine.

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A method for the determination of serum glycated albumin (GA) by high-performance liquid chromatography is presented. The system involves anion exchange chromatography to separate albumin, and consecutive boronate affinity chromatography to separate glycated and nonglycated albumin. Various conditions for measurement with this assay system were examined in comparison with those of fructomamine (FA). The method is precise: CV for within-day assay ranged from 1.29 to 3.77%, and that for between-day assay was 4.94%. Assay of GA by this method was not influcenced by the protein concentration of the samples, or the presence of bilirubin, ascorbic acid or glucose, all of which interferred to some extent with the measurement of FA. Mean recovery, calculated as the percent of added GA recovered in the supplemented samples, was 104.2%(range 99.1-108.7%). Plasma GA concentrations were not significantly different from serum GA values, while plasma FA concentrations were lower than those of serum. The clinical reference ranges for GA and FA were 17.0-23.3% and 2.36-3.05mmol/l, respectively. These were calculated from the determination of 155 healthy subjects based on a probability plot. The GA and FA values of patients with diabetes mellitus were far beyond the respective clinical reference range. The proposed method for the determination of glycated albumin is reliable enough to be applied for clinical use.
一般社団法人日本糖尿病学会の論文