Packed capillary reversed-phase liquid chromatography with high-performance electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry for proteomics

被引：208

作者：

Shen, YF ^{[1
]}

Zhao, R ^{[1
]}

Belov, ME ^{[1
]}

Conrads, TP ^{[1
]}

Anderson, GA ^{[1
]}

Tang, KQ ^{[1
]}

Pasa-Tolic, L ^{[1
]}

Veenstra, TD ^{[1
]}

Lipton, MS ^{[1
]}

Udseth, HR ^{[1
]}

Smith, RD ^{[1
]}

机构：

[1] Pacific NW Natl Lab, Environm Mol Sci Lab, Richland, WA 99352 USA

来源：

ANALYTICAL CHEMISTRY | 2001年 / 73卷 / 08期

关键词：

D O I：

10.1021/ac0011336

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

In this study, high-efficiency packed capillary reversed-phase liquid chromatography (RPLC) coupled on-line with high-performance Fourier transform ion cyclotron resonance (FTICR) mass spectrometry has been investigated for the characterization of complex cellular proteolytic digests. Long capillary columns (80-cm) packed with small (3-mum) C18 bonded particles provided a total peak capacity of similar to 1000 for cellular proteolytic polypeptides when interfaced with an ESI-FTICR mass spectrometer under composition gradient conditions at a pressure of 10 000 psi, Large quantities of cellular proteolytic digests (e.g,, 500 mug) could be loaded onto packed capillaries of 150-mum inner diameter without a significant loss of separation efficiency. Precolumns with suitable inner diameters were found useful for improving the elution reproducibility without a significant loss of separation quality. Porous particle packed capillaries were found to provide better results than those containing nonporous particles because of their higher sample capacity. Two-dimensional analyses from the combination of packed capillary RPLC with high-resolution FTICR yield a combined capacity for separations of >1 million polypeptide components and simultaneously provided information for the identification of the separated components based upon the accurate mass tag concept previously described.

引用

页码：1766 / 1775

页数：10

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