title: A peptide preconcentration approach for nano-high-performance liquid chromatography to diminish memory effects.
abstract: An advanced method has been developed for the analysis of proteolytic digests of complex protein mixtures by reversed phase high-performance liquid chromatography coupled to mass spectrometry. The occurrence of memory effects was prevented by a parallel set of two precolumns employed for simultaneous separation and washing procedures. The system was tested extensively, and tryptic digests of three single proteins were analyzed. In addition, different solvent systems were evaluated for effective washing of the employed precolumns. Using the analytical strategy presented, a reliable identification of proteins in complex mixtures was obtained and not hampered by the occurrence of memory effects.
authors.affiliation: Medical Proteom-Center, Ruhr-University of Bochum, Bochum, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Schaefer
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Chervet
authors.forename: Christian
authors.initials: C
authors.lastname: Bunse
authors.forename: Cornelia
authors.initials: C
authors.lastname: Joppich
authors.forename: Helmut E
authors.initials: HE
authors.lastname: Meyer
authors.forename: Katrin
authors.initials: K
authors.lastname: Marcus
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D010455:Peptides / Q000032:analysis*
chemical_list: D010455:Peptides