Abstract
A colorimetric procedure was developed for the sequential determination of nonenzymatically glycated IgG (GIgG) and albumin (GA). IgG and then albumin were separated from serum or plasma using a single DEAE Cibacron blue F3GA Affi-Gel column. The stable ketoamine linkages present in GIgG and GA reduced a tetrazolium salt to its colored formazan. The method was linear over the range 2.0–25.0% GIgG and 4.0–18.0% GA. For both GIgG and GA, the within- and between-run CV's were <4.5 and 8.5%, respectively, and recoveries were quantitative. The labile aldimine fraction, free glucose, and ascorbate did not affect the results. Nondiabetic reference intervals were 13.2–16.8% GIgG and 6.2–9.7% GA. Nondiabetic and diabetic populations can be clearly discriminated ( P < 0.005). When compared with the thiobarbituric acid assay, the correlation coefficients for GIgG and GA were r = 0.98 and 0.97, respectively. In a mixed group of diabetic and nondiabetic subjects, glycated hemoglobin levels were compared with those of GIgG and GA, and r = 0.68 and 0.35, respectively.
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