Efficient and highly reproducible capillary electrophoresis-mass spectrometry of peptides using Polybrene-poly(vinyl sulfonate)-coated capillaries.
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).
Catai
Jonatan R
JR
Department of Biomedical Analysis, Utrecht University, The Netherlands.
Toraño
Javier Sastre
JS
de Jong
Gerhardus J
GJ
Somsen
Govert W
GW
D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D006583:Hexadimethrine Bromide / Q000737:chemistry*; D013058:Mass Spectrometry / Q000295:instrumentation*; D010455:Peptides / Q000032:analysis*; D011145:Polyvinyls / Q000737:chemistry*; D015203:Reproducibility of Results; D013451:Sulfonic Acids / Q000737:chemistry*
D010455:Peptides; D011145:Polyvinyls; D013451:Sulfonic Acids; C008959:lyapolate; D006583:Hexadimethrine Bromide