Enhanced sequence coverage in tryptic fragment analysis by two-dimensional HPLC/MS using a monolithic silica capillary column |
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| Authors: | Morisaka Hironobu Hata Koji Mima Joji Tanigawa Tetsuya Furuno Masahiro Ishizuka Norio Tanaka Nobuo Ueda Mitsuyoshi |
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| Affiliation: | Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitasshirakawa, Kyoto, Japan. |
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| Abstract: | The HPLC/MS system, in which a monolithic silica capillary column is directly connected to an electronspray-ionization mass spectrometer, showed superior performance at high mobile phase linear velocity. A two-dimensional (2D) HPLC/MS system was established, using an ion-exchange particle-packed capillary column at the first dimension and a monolithic silica capillary column at the second dimension. In an analysis of tryptic fragments from bovine serum albumin, an 81% sequence coverage, obtained by the 2D-HPLC/MS system, increased by 23% as compared to a 1D-HPLC/MS system. This 2D-HPLC/MS system using a monolithic silica capillary column should be useful for enhancing sequence coverage of tryptic fragments in proteomics. |
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