Rapid assay of A1c-type glycosylated hemoglobin in blood of diabetic rats using fast protein liquid chromatography

AIM: To develop a fast protein liquid chromatography (FPLC) method for the assay of A1c-type glycosylated hemoglobin in diabetic rats. METHODS: Venous blood was collected from rats. After the erythrocytes were washed and sedi mented, the hemolysate was prepared and diluted with isotonic saline as a s ample for the assay. Samples were then separated on Mono S HR 5/5 cation ex change column by a lithium chloride gradient elution system. The A1c-type g lycosylated hemoglobin was monitored by measuring the absorbance at 415 nm. RESULTS: The A1c-type glycosylated hemoglobin was well separated from tota l hemoglobin. The average HbA1c amount in diabetic rats was determined to b e (3.6 % +/- 0.6 %, n = 7), while the corresponding figure in normal rats w as (1.4 % +/- 0.4 %, n = 7, P < 0.01). CONCLUSION: This FPLC method is easy , rapid and reproducible, and can be used for an assay of diabetic rat HbA1 c to evaluate and screen new drugs for diabetes mellitus therapy.