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A peptide preconcentration approach for nano‐high‐performance liquid chromatography to diminish memory effects
Author(s) -
Schaefer Heike,
Chervet JeanPierre,
Bunse Christian,
Joppich Cornelia,
Meyer Helmut E.,
Marcus Katrin
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300801
Subject(s) - chromatography , chemistry , mass spectrometry , peptide , high performance liquid chromatography , biochemistry
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.