Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1063, Issue 1-2, Pages 57-69Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2004.11.084
Keywords
proteomics; two-dimensional high performance liquid chromatography (2D-HPLC); electrospray ionization (ESI); mass spectrometry (MS)
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A one- or two-dimensional high performance liquid chromatography system for electrospray ionization mass spectrometers has been developed that is optimized for ion exchange and reversed phase separations. A unique and simple valve configuration permits the use of a variety of non-volatile salts; ammonium sulfate was used in an example of strong cation exchange separations. The system was designed and evaluated for both micro- and nanoflow chromatography. The peptide detection limit was similar to100 fmol for micro- and 20 fmol for nanoflow, demonstrating the concentration and mass sensitivity improvements expected with nanoelectrospray ionization. The 1D/2D-HPLC MS system is fully automated for routine peptide analyses, compatible with direct injection of proteolytic digests, and exhibits chromatographic reproducibility and sensitivity. Software permits operator selection of either a 1D or 2D configuration with corresponding system parameters as required for individual samples. The hardware elements and resulting performance are described in this paper. Published by Elsevier B.V.
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