超高效液相色谱法测定门冬胰岛素含量

Content Determination of Insulin Aspart by UPLC

目的建立测定门冬胰岛素含量的超高效液相色谱法。方法色谱柱为Shim-pack GISS C_(18)柱(100 mm×2.1 mm,1.9μm);流动相A为0.2 mmol/L Na_2SO_4溶液(加85%磷酸溶液2.7 mL,用乙醇胺调pH至3.6)-乙腈(72∶18,V/V),流动相B为50%乙腈,梯度洗脱;流速为0.25 m L/min,检测波长为214 nm,柱温为40℃,进样量为2μL。结果门冬胰岛素质量浓度在0.05～1.30 g/L范围内与峰面积线性关系良好,精密度、稳定性、重复性试验的RSD均小于1%,加样回收率为100.42%,RSD为0.41%(n=9)。结论该方法专属性强,精密度、稳定性、重复性好,可用于门冬胰岛素的含量测定。

Objective To establish a UPLC method for the content determination of insulin aspart.Methods The chromatographic column was Shim-pack GISS C18 column(100 mm×2.1 mm,1.9μm),the mobile phase A was 0.2 mmol/L Na2SO4 solution-acetonitrile(72∶18,V/V),2.7 mL 85%phosphoric acid solution was added,pH was adjusted to 3.6 with ethanolamine,the mobile phase B was 50%acetonitrile with gradient elution,the flow rate was 0.25 mL/min,the detection wavelength was 214 nm,the column temperature was 40℃,and the sample size was 2μL.Results The insulin aspart showed a good linear relationship in the range of 0.05-1.30 g/L.RSDs of precision,stability and repeatability tests were all less than 1%,the average recovery rate was 100.42%,and RSD was 0.41%(n=9).Conclusion The method is specific,accurate,stable and reproducible,which can be used for the content determination of insulin aspart.