色谱

色谱 ›› 2015, Vol. 33 ›› Issue (11): 1155-1162.DOI: 10.3724/SP.J.1123.2015.06021

• 研究论文 • 上一篇    下一篇

一种多通道匀浆装填毛细管色谱柱的新装置及应用

吕雅瑶, 郝斐然, 王欢欢, 付斌, 钱小红, 张养军   

  1. 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:2015-06-16 出版日期:2015-11-08 发布日期:2012-05-30
  • 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(2012YQ12004407,2011YQ030139,2011YQ06008408,2013YQ14040506);国家自然科学基金项目(21275519,20735005,31100591).

A novel multiple-channel apparatus for packing capillary chromatographic column and its application

LV Yayao, HAO Feiran, WANG Huanhuan, FU Bin, QIAN Xiaohong, ZHANG Yangjun   

  1. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China
  • Received:2015-06-16 Online:2015-11-08 Published:2012-05-30

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摘要：

针对目前毛细管色谱柱装柱效率低、不同批次装填的毛细管色谱柱之间性能差异大的问题,我们发展了一种多通道匀浆装填毛细管色谱柱的新装置。该装置以液相色谱泵提供压力、采用磁力搅拌保持匀浆液均匀分散,一次可装填多达6根毛细管色谱柱。以牛血清白蛋白(BSA)的胰蛋白酶酶切肽段混合物为样本,选择峰容量、蛋白覆盖率、3个特定离子的保留时间以及毛细管色谱柱柱压为指标,在毛细管液相色谱-质谱联用系统上对装填的反相毛细管色谱柱的性能进行了评价。分别考察了一次装填的6根毛细管色谱柱、两次装填的12根毛细管色谱柱以及一次装填1根与一次装填6根毛细管色谱柱的性能及稳定性。实验结果表明:同一批次装填的6根毛细管色谱柱的性能相近;不同批次装填的12根毛细管色谱柱的峰容量和覆盖率没有明显的区别,但保留时间和毛细管色谱柱柱压的稳定性较差;一次装填1根和一次装填6根毛细管色谱柱柱性能的稳定性与两次分别装填6根毛细管色谱柱的稳定性相近,即采用本装置可显著提高毛细管色谱柱的装填效率且每次装填毛细管色谱柱的数量不会对柱性能产生影响。

关键词: 多通道, 毛细管色谱柱, 毛细管液相色谱-质谱联用, 装柱装置

Abstract:

A novel multiple-channel apparatus for packing capillary chromatographic column was designed and manufactured for packing six capillary chromatographic columns with close column efficiency at the same time. Briefly, it consists of a magnetic stirrer, a liquid chromatographic pump and a multiple-channel can. The reagents used for preparing ODS (C18) slurry and stirring condition of the magnetic stirrer were optimized in the study. Two batches of capillary chromatographic columns were packed under the optimum condition, and these packed capillary chromatographic columns were evaluated in the terms of peak capacity, sequence coverage, retention times of three peptide ions and column pressure using the tryptic digest of a bovine serum albumin (BSA) and detected by LC-MS in electrospray ionization (ESI) mode. The experimental results showed that the six capillary chromatographic columns packed at the same time had close column efficiencies, however, the column efficiencies of twelve capillary chromatographic columns packed at two times were significantly different. In addition, there was no significant column efficiency difference when packing one or six capillary chromatographic columns at the same time. The multiple-channel apparatus designed by us is simple, time-saving, and can be applied to pack capillary chromatographic columns with similar column efficiencies, thus it is of evident advantage over traditional one-channel apparatus.

Key words: capillary chromatographic column, capillary liquid chromatography-mass spectrometry (CLC-MS), multiple-channel, packing column apparatus

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