高效液相色谱法测定胰岛素及其制剂中胰岛素的效价

目的建立用高效液相色谱法测定胰岛素及其制剂的效价,并将之与已有的小鼠血糖法比较。方法色谱柱:C18柱(岛津VP-ODS,4.6 mm×150 mm,5μm);流动相:0.2 mol/L硫酸盐缓冲液(用乙醇胺调pH至2.3)-乙腈(73∶27);检测波长:214 nm;流速:1 mL/min。结果胰岛素在10~50 u/mL范围内线性关系良好,回归方程为:Y=0.001 4X+0.222 7,r=0.999 8;精密度、重现性良好。结论此法灵敏度高、专属性强、准确、可靠,较生物测定法方便,适于胰岛素及其制剂中胰岛素的效价测定。

Determination of the insulin potency in insulin and its preparations by HPLC

Purpose To develop a HPLC method for the determination of insulin potency and to compare it with the method to assay the potency based on the glucose content in mice′s blood.Methods The separation was performed on a VP-ODS column(4.6 mm×150 mm,5 μm).The mobile phase was composed of 0.2 mol/L sodium sulfate(pH was adjusted to 2.3 with ethanolamine)-acetonitrile(73∶27) with flow rate 1 mL/min,detected at 214 nm. Results The calibration curve was linear in the range of 10-50 u/mL.The regression equation of insulin was Y=0.001 4 X+0.222 7,r=0.999 8(n=5).Conclusion The HPLC method was simple,sensitive and reliable for the determination of insulin.