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目的:建立快速测定糖尿病大鼠Alc型糖基化血红蛋白的FPLC方法。方法:大鼠血样经洗涤,离心,甲苯破膜,稀释后;用丙二酸钠溶液(A液)和丙二酸钠-氯化钾溶液(B液)为基本流动相,采用梯度洗脱程序在Mono S HR 5/5阳离子交换柱上进行分离测定,检测波长为415nm。结果:在所给色谱条件下,大鼠Alc型糖基化血红蛋白能很好地从总血红蛋白中分离出来。糖尿病大鼠中HbAlc含量为3.6％±0.6％(n=7)显著高于正常大鼠的1.4％±0.4％(n=7),P<0.01。测定一个样品所需的总时间约为27min。结论:本方法简便,快速,准确,重现性好,可用于大鼠糖尿病模型中糖基化血红蛋白含量的研究和糖尿病治疗药物的筛选。
Objective: To establish a rapid FPLC method for the determination of A1c glycosylated hemoglobin in diabetic rats. Methods: The blood samples of rats were washed, centrifuged and broken through the membrane of toluene. After dilution, sodium molybdate solution (A solution) and sodium malonate solution - potassium chloride solution (B solution) The program was run on a Mono S HR 5/5 cation exchange column with a detection wavelength of 415 nm. Results: Rat Alc glycated hemoglobin was well separated from the total hemoglobin under the chromatographic conditions. The HbA1c level in diabetic rats was 3.6% ± 0.6% (n = 7), significantly higher than that in normal rats (1.4% ± 0.4%, n = 7) (P <0.01). The total time required to determine a sample is about 27 min. Conclusion: The method is simple, rapid, accurate and reproducible. It can be used to study the content of glycosylated hemoglobin in diabetic model of rats and the screening of diabetes therapeutic drugs.