Original Article

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

Authors: Hui Ge, Xiao-Ming Li, Zhen-Chuan Miao, Wei Song, Shu-Sen Liu


To develop a fast protein liquid chromatography (FPLC) method for the assay of Alc-type glycosylated hemoglobin in diabetic rats.
Venous blood was collected from rats. After the erythrocytes were washed and sedimented, the hemolysate was prepared and diluted with isotonic saline as a sample for the assay. Samples were then separated on Mono_S_HR_5/5 cation exchange column by a lithium chloride gradient elution system. The Alc-type glycosylated hemoglobin was monitored by measuring the absorbance at 415 nm.
The Alc-type glycosylated hemoglobin was well separated from total hemoglobin. The average HbAlc amount in diabetic rats was determined to be (3.6% +/- 0.6%, n = 7), while the corresponding figure in normal rats was (1.4% +/- 0.4%, n = 7, P < 0.01).
This FPLC method is easy, rapid and reproducible, and can be used for an assay of diabetic rat HbAlc to evaluate and screen new drugs for diabetes mellitus therapy."

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