色谱

色谱 ›› 2015, Vol. 33 ›› Issue (6): 563-570.DOI: 10.3724/SP.J.1123.2015.01036

• 研究论文 • 上一篇    下一篇

一种简单的毛细管色谱柱电加热装置的制作方法及其在液相色谱-质谱联用系统中的应用

金祖耀1,2, 吕雅瑶2, 周珊珊2, 郝斐然2, 付斌2, 应万涛2, 钱小红2, 张养军1,2   

  1. 1. 安徽医科大学, 安徽 合肥 230032;
    2. 蛋白质组学国家重点实验室, 北京蛋白质组研究中心, 军事医学科学院放射与辐射医学研究所, 北京 102206
  • 收稿日期:2015-01-27 出版日期:2015-06-08 发布日期:2015-05-26
  • 通讯作者: 张养军
  • 基金资助:

    国家重大科学计划项目(2012CB910603);国家高技术研究发展规划项目(2012AA020202);国家重大科学仪器设备开发专项项目(2011YQ09000504,2012YQ12004407,2011YQ030139,2011YQ06008408);国家自然科学基金项目(21275519,20735005,31100591).

A simple preparation method of an electric heating apparatus for heating capillary chromatographic columns and its application in liquid chromatography-mass spectrometry system

JIN Zuyao1,2, LÜ Yayao2, ZHOU Shanshan2, HAO Feiran2, FU Bin2, YING Wantao2, QIAN Xiaohong2, ZHANG Yangjun1,2   

  1. 1. Anhui Medical University, Hefei 230032, China;
    2. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China
  • Received:2015-01-27 Online:2015-06-08 Published:2015-05-26

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摘要：

为了实现蛋白质组的深度覆盖,特别是低丰度蛋白质的定性鉴定和定量分析,目前常用的方法是采用更长或装填更小粒径填料的毛细管色谱柱,但因此带来的问题是色谱柱反向柱压显著升高。针对以上问题发展了一种简单的毛细管色谱柱电加热装置制作方法,并将该装置安装于液相色谱-质谱联用系统,分别以牛血清白蛋白(BSA)酶切肽段混合物和酵母蛋白(yeast)酶切肽段混合物为样品,从柱压和柱效两方面对该装置的性能进行了评价。实验结果表明,所制作的毛细管柱电加热装置安装在装填粒径为3 μm反相色谱填料的毛细管柱上,在最佳电流(100 mA)下对BSA及yeast酶切肽段混合物进行分离时的柱压比不加电流时的柱压降低至少50%,柱效略有升高。这说明所制作的毛细管色谱柱电加热装置能显著降低柱压,为在较低的柱压条件下选择更小粒径色谱颗粒填料的毛细管色谱柱提供了一种有效的方法。

关键词: 蛋白质组学, 电加热装置, 毛细管高效液相色谱-串联质谱, 柱效, 柱压

Abstract:

For deep coverage of proteome, especially in performing qualitative identification and quantitative analysis of low-abundance proteins, the most commonly used method is the application of a longer capillary chromatographic column or a capillary column packed with smaller particle sizes. However, this causes another problem, the very high back pressure which results in liquid leaks in some connection parts in a liquid chromatograph. To solve this problem, an electric heating apparatus was developed to raise the temperature of a capillary column for reducing its back pressure, which was further applied in a capillary high performance liquid chromatography-tandem mass spectrometry system (cHPLC-MS/MS), and evaluated in the terms of chromatographic column back pressure and chromatographic column efficiency using bovine serum albumin (BSA) tryptic digests and yeast tryptic digests, separately. The results showed that at the optimum current, our electric heating apparatus could reduce the column pressure of a capillary column packed with 3 μm packing materials by at least 50% during the separation of BSA tryptic digestion and yeast tryptic digestion, compared with that without electric heating. The column efficiency was also increased slightly. This suggested that the electric heating apparatus can significantly reduce the column pressure, which provides an efficient way to use capillary chromatographic columns packed with smaller sizes of particles at a lower pressure.

Key words: capillary high performance liquid chromatography-tandem mass spectrometry (cHPLC-MS/MS), column efficiency, column pressure, electric heating apparatus, proteomics

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