Abstract
We investigated the impact of one dimension (single reverse phase (RP) column) and two dimension (two different RP columns) chromatographic methods on SIM (MS) and multiple reaction monitoring (MRM; MS/MS) performance from human plasma. We find that MRM analysis is clearly preferable for 1-D applications; however, implementation of SIM detection in conjunction with 2-D separation technique resulted in an over 60-fold increase in analyte peak area and improved S/N compared to MRM for our analyte, human C-peptide. Implementation of a 2-D RP-RP technique with SIM detection is capable of eliminating matrix effects and greatly increases signal response and data quality. For two large peptides in complex biological samples, we found that a 2-D approach performed better than high quality sample preparation together with 1-D chromatography and MRM, even on a high-end mass spectrometer.
Original language | English (US) |
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Pages (from-to) | 226-233 |
Number of pages | 8 |
Journal | Journal of Separation Science |
Volume | 30 |
Issue number | 2 |
DOIs | |
State | Published - Feb 2007 |
Keywords
- C-peptide
- Column switching
- Ionization efficiency
- Mass spectrometry
- Matrix effects
ASJC Scopus subject areas
- Analytical Chemistry
- Filtration and Separation