色谱

色谱

• 研究论文 • 上一篇    下一篇

反相毛细管整体柱的制备及其在多肽混合物分离中的应用

谢晶鑫1,毕开顺1,钱小红2,张养军2*   

  1. 1.Shenyang Pharmaceutical University, Shenyang 110015, China; 2.Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China
  • 收稿日期:2008-09-10 修回日期:2008-11-10 出版日期:2009-03-30 发布日期:1982-12-25
  • 通讯作者: 张养军
  • 基金资助:
    国家重点基础研究规划项目(2006CB910803, 2007CB914104, 2004CB518707)、 国家高技术研究发展计划项目(2006AA02A308)和国家自然科学基金项目(30621063, 20635010, 20735005, 20875101)

Preparation of a reversed-phase capillary monolithic column and its application in the separation of polypeptide mixtures

XIE Jingxin1, BI Kaishun1, QIAN Xiaohong2, ZHANG Yangjun2*   

  1. 1.Shenyang Pharmaceutical University, Shenyang 110015, China; 2.Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China
  • Received:2008-09-10 Revised:2008-11-10 Online:2009-03-30 Published:1982-12-25
  • Contact: ZHANG Yangjun

PDF

1068

被引次数

7

摘要： 采用甲基丙烯酸月桂酯为基础功能单体,乙二醇二甲基丙烯酸酯为交联剂,正十二醇、1,4-丁二醇及二甲基亚砜为致孔剂,在内径为75 μm的石英毛细管内制备了具有良好机械性能及化学稳定性的反相毛细管整体柱。考察了致孔剂的种类、比例以及交联剂在单体混合物中的比例对柱压和分离效果的影响;以单体15%、交联剂15%、致孔剂70%(均为质量分数)作为优化配方,在70 ℃条件下反应24 h;并对所合成的毛细管整体柱进行了电镜表征,测试了流速、柱长与柱压的关系。结果表明，毛细管整体柱的通透性良好,可通过延长柱长的方法提高分离效果。将所制备的毛细管整体柱装于纳升级高效液相色谱仪上进行牛血清白蛋白及血浆样本的胰蛋白酶酶切液的分离,获得了比较理想的分离效果。

关键词: 多肽分离 , 毛细管整体柱, 纳升级液相色谱

Abstract: Capillary monolithic columns (75 μm i.d.) were prepared by the copolymerization of lauryl methacrylate as the basic monomer, ethylene dimethacrylate as the cross-linking agent and 1-dodecyl alcohol, 1,4-butanediol and dimethyl sulfoxide as the porogenic mixture. The synthetic stationary phases had better mechanical properties and chemical stabilities. A series of characterization and evaluations were performed on the capillary monolithic columns including the scanning electron microscope (SEM) images, the influences of pressure and the effects on the separation of peptide mixtures by changing the proportions of the porogen solution and cross-linking agent. The final prescription contained 15% (w/w) monomer, 15% (w/w) cross-linking agent, and 70% (w/w) porogenic agent. Then the solution was heated at 70 ℃ for 24 h. The test of relationship between column length and back pressure showed that the capillary monolithic columns prepared have superior permeability, so a longer column can be used to improve the effects of separation. The prepared capillary monolithic columns are fitted on the nano-scale high performance liquid chromatography for the separation of tryptic digests of bovine serum albumin (BSA) and human plasma samples, and better results have been obtained.

Key words: nano-scale high performance liquid chromatography (nano-HPLC), polypeptide separation , capillary monolithic column