Capillary high-performance liquid chromatography-electrospray ionization mass spectrometry using monolithic columns and carbon fiber electrospray ionization emitters.

Kyung Won Ro; Jian Liu; Mark Busman; Daniel R Knapp

doi:10.1016/j.chroma.2004.06.115

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Capillary high-performance liquid chromatography-electrospray ionization mass spectrometry using monolithic columns and carbon fiber electrospray ionization emitters.

Kyung Won Ro·Jian Liu·Mark Busman·Daniel R Knapp

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Abstract

Monolithic columns having long hydrocarbon chains were prepared by in-situ polymerization in capillary fused silica tubing. The capillary columns were coupled with a newly developed carbon fiber electrospray ionization (ESI) emitter for proteomic analysis using sheathless capillary HPLC-ESI mass spectrometry (MS). The sample loading capacity and chromatographic performance of the styrene-based monolithic column, which was prepared by photo-polymerization of octylstyrene (OS) and divinylbenzene (DVB) were compared with that of the methacrylate-based monolithic column composed of lauryl methacrylate (LMA) and ethylene dimethacrylate (EDMA). The sample loading ability of tryptic digested protein in poly-OS (POS)-DVB column was higher than that of poly-LMA (PLMA)-EDMA column, possibly due to the irregular and rugluous surface offering a greater surface area of POS-DVB stationary phase. The POS-DVB column also provided better separation efficiency in the separation of high concentration (10 microg) of tryptic digested albumin bovine serum (BSA). Due to the successful interface of a highly efficient monolithic column and a stable, durable carbon fiber emitter, low femtomole levels of peptides were successfully separated and identified in the presence of large amounts of tryptic digested protein.

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Amino Acid Sequence Carbon Chromatography, High Pressure Liquid Microscopy, Electron, Scanning Molecular Sequence Data Myoglobin Sensitivity and Specificity Spectrometry, Mass, Electrospray Ionization Trypsin Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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Kyung Won Ro, Jian Liu, Mark Busman, Daniel R Knapp. 2004-08-20. Capillary high-performance liquid chromatography-electrospray ionization mass spectrometry using monolithic columns and carbon fiber electrospray ionization emitters.. https://doi.org/10.1016/j.chroma.2004.06.115

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