小样本量预混精蛋白重组人胰岛素注射液的效价测定

目的: 建立小样本量(<1 mL)预混精蛋白重组人胰岛素注射液的效价检测方法,并进行相关的方法学验证。 方法: 采用C₁₈柱(Phenomenex,4.6 mm×250 mm,5 μm);流动相为0.2 mol·L^-1硫酸盐缓冲液-乙腈(73∶ 27);检测波长为214 nm;流速1.0 mL·min^-1;柱温40 ℃。 结果: 重组人胰岛素溶液在6 h内稳定性良好,低、中、高浓度组的加样回收率分别为96.7%,97.0%,96.7%,小样本量法与大样本量法(即中国药典规定方法)检测同批次预混精蛋白重组人胰岛素注射液效价,结果无显著区别。 结论: 本文建立的小样本量法适用于评价单只预混精蛋白重组人胰岛素笔芯及使用中笔芯的实际效价,对检测具有重要的实用价值。

Objective: An HPLC method for the determination of pre-mixed isophane protamine recombinant human insulin injection with small sample size (<1 mL) was established and the methodology validation was carried out at the same time. Method: C₁₈ column (Phenomenex,4.6 mm×250 mm,5 μm) was used.The mobile phase was sulfate buffer (0.2 mol·L^-1 )-acetonitrile (73∶ 27) and the flow rate was 1.0 mL·min^-1,the detection wavelength at 214 nm. Results: The solution of recombinant human insulin was stable within 6 hours.The results for the average recovery rates of low,middle and high concentration groups were 96.7%,97.0% and 96.7%.The same batch of pre-mixed isophane protamine recombinant human insulin injection were tested,the results of small sample size method and the large sample size method (ChP method) showed no significant difference. Conclusion: Small sample size method is suitable for the evaluation of single cartridge and the actual titer of single using cartridge.