On-Column Sample Enrichment for Capillary Electrophoresis Sheathless Electrospray Ionization Mass Spectrometry: Evaluation for Peptide Analysis and Protein Identification
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- 作者：George M. Janini ; Ming Zhou ; Li-Rong Yu ; Josip Blonder ; Michelle Gignac ; Thomas P. Conrads ; Haleem J. Issaq ; and Timothy D. Veenstra
- 刊名：Analytical Chemistry
- 出版年：2003
- 出版时间：November 1, 2003
- 年：2003
- 卷：75
- 期：21
- 页码：5984 - 5993
- 全文大小：267K
- 年卷期：v.75,no.21(November 1, 2003)
- ISSN：1520-6882
文摘
Although several designs have been advanced for couplingsample enrichment devices to a sheathless electrosprayionization-mass spectrometry (MS) interface on a capillaryelectrophoresis (CE) column, most of these approachessuffer from difficulties in fabrication, and the CE separation efficiency is degraded as a result of the presence ofcoupling sleeves. We have developed a design that offerssignificant improvements in terms of ease of fabrication,durability, and maintenance of the integrity of the CE-separated analyte zones. Capillaries with different insideand outside diameters were evaluated to optimize theperformance of the CE-MS system, resulting in a masslimit of detection of 500 amol for tandem MS analysis ofa standard peptide using a 20-
m-i.d. capillary. Theimproved design incorporates an efficient method topreconcentrate a sample directly within the CE capillaryfollowed by its electrophoretic separation and detectionusing a true zero dead-volume sheathless CE-MS interface. Testing of this novel CE-MS system showed itsability to characterize proteomic samples such as proteindigests, in-gel-digested proteins, and hydrophobic peptides as well as to quantitate ICAT-labeled peptides.
m-i.d. capillary. Theimproved design incorporates an efficient method topreconcentrate a sample directly within the CE capillaryfollowed by its electrophoretic separation and detectionusing a true zero dead-volume sheathless CE-MS interface. Testing of this novel CE-MS system showed itsability to characterize proteomic samples such as proteindigests, in-gel-digested proteins, and hydrophobic peptides as well as to quantitate ICAT-labeled peptides.