Two-Dimensional SEC/RPLC Coupled to Mass Spectrometry for the Analysis of Peptides
Citations Over TimeTop 10% of 1997 papers
Abstract
A two-dimensional liquid chromatography system is described here which uses size exclusion liquid chromatography (SEC) followed by reversed phase liquid chromatography (RPLC) to separate the mixture of peptides resulting from the enzymatic digestion of a protein. A novel LC/LC interface, using two RPLC columns in parallel rather than storage loops, joins the two chromatographic dimensions. This new interface design permits the use of conventional analytical diameter HPLC columns, 7.8 mm for SEC and 4.6 mm for RPLC, making construction and maintenance of this system very easy. The reversed phase chromatography utilizes 1.5 microns diameter, nonporous C-18 modified silica particles, which produce fast and efficient analyses. Following the high-resolution two-dimensional chromatographic separation, an electrospray mass spectrometer detects the peptide fragments. The mass spectrometer scans a 2000 m/z range to identify the analytes from their molecular weights. The analyses of tryptic digests of ovalbumin and serum albumin are each described.
Related Papers
- → Applications of Tandem Mass Spectrometry (MS/MS) in Protein Analysis for Biomedical Research(2022)129 cited
- → Electrospray Mass Spectrometry and Tandem Mass Spectrometry of Synthetic Multicomponent Peptide Mixtures: Determination of Composition and Purity(1994)127 cited
- → 23rd Sanibel Conference on Mass Spectrometry: From Fragmentation Mechanisms to Sequencing: Tandem Mass Spectrometry Based Peptide and Protein Identification(2012)2 cited
- → Application of electrospray and fast atom bombardment mass spectrometry to the identification of post-translational and other chemical modifications of proteins and peptides(1994)14 cited
- Analysis of the Primary Structure of Modified and Unknown Peptides by Nanoelectrospray Tandem Mass Spectrometry(2007)