Abstract
We have revised current two-dimensional RP-RP approaches and developed a new robust 2-D RP-RP platform. This platform was implemented on an Agilent 1100 2-D liquid chromatography system and is based on high pressure switching between two high-resolution RP columns. An independent binary gradient was implemented for each dimension. The powerful combination of dual analytical columns with independent gradient elution achieves high analyte purity, effectively eliminates matrix effects, and maximizes MS sensitivity in Q1 SIM comparable to the sensitivity enhancements of MS/MS-based methods. Implementation of dual simultaneous gradient profiles (overlapped gradients) reduces 2-D method run-time to the scale of 1-D method run-times. This robust and sensitive approach is particularly suitable for hydrophobic peptides and small proteins and can be used as a routine standard technique for enhanced on-line peptide purification coupled with mass spectrometric detection.
Original language | English (US) |
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Pages (from-to) | 538-546 |
Number of pages | 9 |
Journal | Journal of Separation Science |
Volume | 29 |
Issue number | 4 |
DOIs | |
State | Published - Mar 2006 |
Keywords
- Carryover
- Column switching
- Mass spectrometry
- Matrix effects
- Peak focusing
ASJC Scopus subject areas
- Analytical Chemistry
- Filtration and Separation