Determination of glycosylated hemoglobin by affinity chromatography

Conclusions

The described new affinity chromatography method for the determination of glycosylated hemoglobins is rapid, specific and precise and has several advantages over other methods. It is relatively insensitive to changes in temperature and pH and appears to be free of interferences by labile Glyc-Hb. The results are independent over a wide range of hemolysate concentrations. The method offers good stabilities of specimen, buffers and columns. The possibility of a 10-fold reuse and a simple sample preparation by whole blood hemolysis without washing or incubation of erythrocytes make this method technically less complicated and more suited for routine use in a clinical laboratory.

Correlation studies between affinity method and ion-exchange method show that as the level of glycosylation increases the affinity method gives progressively higher values (factor 1.3–1.4 in the pathological range). These results indicate that the affinity method measures not only theβ-terminal modified Hb molecules but all glycosylated hemoglobins, thus giving a better discrimination between normoglycemic and diabetic samples.