Peptide enrichment by microfluidic electrocapture for online analysis by electrospray mass spectrometry.
Identification of peptides from a complex mixture can be difficult because of the wide concentration range and the different ionization efficiencies of peptides during analysis by electrospray ionization (ESI) mass spectrometry (MS). Preconcentration methods are necessary to allow low-abundance and low-intensity peptides to reach the ionization threshold of the mass spectrometer. Here we demonstrate peptide enrichment based on electroimmobilization. Peptides are immobilized without the use of solid support or chemical binding by application of an electric field along a microflow stream in an electrocapture cell. Once enriched/preconcentrated inside the cell, they are released by removal of the electric field and via an interface with an electrospray emitter are submitted to online mass spectrometric analysis. Tandem mass spectrometric analysis of a peptide mixture containing hemoglobin, myoglobin, bovine serum albumin (BSA), and cytochrome c was successful. Amplification factors up to 16-fold were achieved with improvement of the signal-to-noise values for the preconcentrated sample. The limit of detection for one of the preconcentrated peptides was 3.6 fmol.
Vollmer
Susanne
S
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-171 77 Stockholm, Sweden.
Astorga-Wells
Juan
J
Alvelius
Gunvor
G
Bergman
Tomas
T
Jörnvall
Hans
H
D046210:Microfluidic Analytical Techniques / Q000379:methods*; D010455:Peptides / Q000302:isolation & purification*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D010455:Peptides