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重组人钙调磷酸酶B亚基的毛细管区带电泳分析方法

Determination of Recombinant Human Calcineurin B Subunit by Capillary Zone Electrophoresis

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【作者】 高锦黄晨西魏群

【Author】 GAO Jin, HUANG Chen - xi, WEI Qun ( Department of Biochemistry and Molecular Biology,Beijing Normal University,Beijing 100875 ,China; Institute for Drug and Instrument Control of Shenyang Military Command of PLA, Shenyang 110026, China)

【机构】 北京师范大学生化与分子生物学系

【摘要】 目的:建立毛细管区带电泳(CZE)分离分析重组人钙调磷酸酶B亚基的新方法。方法:用毛细管区带电泳分离模式, 采用未涂层熔融石英毛细管50μm×57 cm;电泳缓冲液:0.2 mol·L-1 2-吗啉代乙磺酸-0.05 mol·L-1柠檬酸盐-乙腈 (5:5:1,pH 2.5);分离电压:15 kV;柱温:25℃;压力进样5s;检测波长:214 nm,进行纯度分析。结果:所建立的CZE分析方法能够将重组人钙调磷酸酶B亚基与未知杂质有效分离,主峰的迁移时间及峰面积的RSD分别为0.31%和1.5%。结论:所建立的毛细管区带电泳方法可以用于重组人钙调磷酸酶B亚基的纯度分析,它具有高效、快速、简便及样品和试剂消耗少等优点。

【Abstract】 Objective:To establish a new capillary zone electrophoresis(CZE) method for the analysis of recombinant human calcineurin B subunit(rhCNB). Method:Based on CZE mode to determinate purity. On uncoated fused silica capillary 50μm×57 cm, using 0. 2 mol·L-1 2 - morpholinoethanesulfonic acid monohydrate - 0. 05 mol·L-1 citrate - acetonitrile(5:5:1 ,pH 2.5) as electrolytes. Electrophoresis was performed at 15 kV and 25℃. The sample was introduced into the capillary by pressure injection for 5 s. The wavelength of detection was 214 nm. Re-sults:rhCNB and unknown impurity were effectively separated by the established method. The RSD of migration time and peak area were 0. 31% and 1. 5% respectively. Conclusion: The established CZE method can be utilized for the purity analysis of rhCNB. It has the advantages of accuracy, fastness, convenience, low sample and reagent consumption.

【基金】 国家863计划(2002AA214131)资助项目国家自然科学基金(39930060)资助项目
  • 【文献出处】 药物分析杂志 ,Chinese Journal of Pharmaceutical Analysis , 编辑部邮箱 ,2005年07期
  • 【分类号】R917
  • 【被引频次】1
  • 【下载频次】65
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