Premium
Efficient and highly reproducible capillary electrophoresis–mass spectrometry of peptides using Polybrene‐poly(vinyl sulfonate)‐coated capillaries
Author(s) -
Catai Jonatan R.,
Toraño Javier Sastre,
de Jong Gerhardus J.,
Somsen Govert W.
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500915
Subject(s) - chemistry , chromatography , capillary electrophoresis , formic acid , bilayer , coating , sulfonate , mass spectrometry , capillary electrophoresis–mass spectrometry , capillary action , polymer , analytical chemistry (journal) , electrospray ionization , materials science , membrane , biochemistry , organic chemistry , composite material , sodium
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).