重组人C1酯酶抑制剂中rcc-rhC1INH和ox-rhC1INH含量检测

Determination of rcc-rhC1INH and ox-rhC1INH in the recombinant human C1 esterase inhibitor

目的:分析重组人C1酯酶抑制剂(rhC1INH)中相关蛋白rcc-rhC1INH和ox-rhC1INH的含量。方法:采用反相高效液相色谱法进行测定,对该方法进行专属性、准确性和精密度检验,并用该方法对4批样品中的相关蛋白进行检测。结果:低浓度下,rhC1INH对照品、rcc-rhC1INH对照品和ox-rhC1INH对照品能够较好分离。rcc-rhC1INH和ox-rhC1INH 2种对照品质量浓度为0.2、1和1.4 mg·mL^(-1)时的回收率均在85%～115%的范围内。对同1批3个待测样品进行测定,rcc-rhC1INH含量为(0.082±0.008)%,RSD为9.8%;ox-rhC1INH含量为(0.35±0.05)%,RSD为14.3%,显示该方法具有较好的专属性、准确性和精密度。4批测试样品中,rcc-rhC1INH的含量分别为0.083%、0.021%、0.033%、0.042%,ox-rhC1INH的含量分别为0.34%、0、0、0。结论:经方法学验证,RP-HPLC方法可用于rhC1INH中rcc-rhC1INH和ox-rhC1INH的含量检测。

Objective：To analyze the contents of related proteins rcc-C1 INH and ox-C1 INH in the recombinant human C1 esterase inhibitor（rhC1 INH）. Methods：The reversed phase high-performance liquid chromatography（RP-HPLC） was used to determine the relative proteins,the test was verified in specificity,accuracy and precision,and was used to detect the rcc-C1 INH and ox-C1 INH in 4 batches of samples. Results：RhC1 INH reference,rcc-rh C1 INH reference and ox-rhC1 INH reference could be well separated under the low concentrations.The recovery rates for the reference substances of rcc-C1 INH and ox-C1 INH at three concentrations（0.2,1 and 1.4 mg·mL-1） were all in the range of 85%～115%. Three groups from one batch were determined separately,the contents of rcc-C1 INH were（0.082±0.008）%,and the contents of ox-C1 INH were（0.35±0.05）%,with RSDs of 9.8% and 14.3%,respectively. This method showed good results in specificity,accuracy and precision. Four batches of rhC1 INH were determined,the contents of rcc-C1 INH were 0.083%,0.021%,0.033% and 0.042% for each,and ox-C1 INH were 0.34%,0,0,and 0,respectively. Conclusion：RP-HPLC can be used for the content determination of rcc-rhC1 INH and ox-rhC1 INH in rhC1 INH.