Koulich D M, Nazimov I V, Maltsev K V, Wulfson A N
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow.
J Chromatogr B Biomed Appl. 1994 Dec 9;662(2):357-62. doi: 10.1016/0378-4347(94)00219-3.
The optimization of the separation of recombinant human insulin (rhI) and recombinant human proinsulin (rhP) by free-solution capillary zone electrophoresis in uncoated fused-silica capillaries with ionic and zwitterionic buffers was carried out. The relationship between the selectivity and pH of the buffer was established. The effects of pH and ionic strength of the buffer on protein adsorption on the capillary walls and Taylor diffusion was investigated. The separation of rhI and rhP with an efficiency of 200,000 theoretical plates was achieved using 20 mM Na2HPO4-NaOH buffer (pH 11.2). The proposed method allows the simultaneously determination of rhP and some rhI degradation products (which are also included in pharmacopoeias) in pharmacopoeially limited amounts in rhI.
采用离子缓冲液和两性离子缓冲液，在未涂层熔融石英毛细管中通过自由溶液毛细管区带电泳对重组人胰岛素（rhI）和重组人胰岛素原（rhP）的分离进行了优化。建立了缓冲液选择性与pH值之间的关系。研究了缓冲液的pH值和离子强度对蛋白质在毛细管壁上的吸附以及泰勒扩散的影响。使用20 mM Na2HPO4-NaOH缓冲液（pH 11.2）实现了rhI和rhP的分离，分离效率达到200,000理论塔板数。所提出的方法能够同时测定rhI中符合药典限量的rhP和一些rhI降解产物（这些产物也包含在药典中）。
