Abstract
The potential of capillaries noncovalently coated with a bilayer of oppositely charged polymers for the analysis of peptides by CE-MS was investigated. Bilayer coatings were produced by subsequently rinsing fused-silica capillaries with a solution of Polybrene (PB) and poly(vinyl sulfonate) (PVS). The PB-PVS coating showed to be fully compatible with MS detection causing no ionization suppression or background signals. The bilayer coating provided a considerable EOF at low pH, thereby facilitating the fast separation of peptides using a BGE of formic acid (pH 2.5). Under optimized CE-MS conditions, for enkephalin peptides high separation efficiencies were obtained with plate numbers in the range of 300 000-500 000. It is demonstrated that both the cancellation of the hydrodynamic capillary flow induced by the nebulizer gas and a sufficiently high-data acquisition rate are crucial for achieving these efficiencies. The overall performance of the CE-MS system using PB-PVS-coated capillaries was evaluated by the analysis of a tryptic digest of cytochrome c. The system provided an efficient separation of the peptide mixture, which could be effectively monitored by MS/MS detection allowing identification of at least 13 peptides within a time interval of 1.5 min. In addition, the PB-PVS coating proved to be very consistent yielding stable CE-MS patterns with highly favorable migration time reproducibilities (RSDs <1% over a 3-day period).
Original language | English |
---|---|
Pages (from-to) | 2091-2099 |
Number of pages | 9 |
Journal | Electrophoresis |
Volume | 27 |
Issue number | 11 |
DOIs | |
Publication status | Published - Jun 2006 |
Keywords
- capillary electrophoresis
- electrospray ionization
- mass spectrometry
- noncovalent bilayer coatings
- peptides
- BILAYER-COATED CAPILLARIES
- ELECTROSPRAY-IONIZATION
- BASIC-PROTEINS
- SEPARATION
- COATINGS
- CHROMATOGRAPHY
- ADSORPTION