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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

ISSN号：1000-8713
期刊名称：《色谱》
时间：0
分类：O658[理学—分析化学;理学—化学]
作者机构：[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
相关基金：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）

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

关键词：毛细管高效液相色谱-串联质谱, 蛋白质组学, 电加热装置, 柱压, 柱效, capillary high performance liquid chromatography-tandem mass spectrometry（cHPLC-MS/MS） , proteomics , electric heating apparatus , column pressure , column efficiency

中文摘要：

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3μm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。

英文摘要：

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 perform-ance liquid chromatography-tandem mass spectrometry system（ cHPLC-MS/MS）,and evalua-ted in the terms of chromatographic column back pressure and chromatographic column effi-ciency 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.