Optimization of experimental parameters for electron capture dissociation of peptides in a Fourier transform mass spectrometer.
This paper describes our effort in optimizing the experimental parameters for electron capture dissociation (ECD) of peptides in a commercially available Fourier-transform mass spectrometer. Using a built-in electrically heated filament electron gun, it was demonstrated that good quality ECD spectra of peptides (MW < 2500) could be obtained by irradiating the isolated peptide molecule-ions with a short pulse (50 ms) of low-energy (3-6 eV) electrons. In addition, we have also demonstrated that pulsing of inert cooling gas (argon) could further improve the intensity of the ECD-induced fragment ions. Due presumably to the influence of the strong magnetic field on the trajectories of electrons, the distance between the electron gun and the trapped-ion cell (i.e., 108 mm versus 20 mm) was found to have little influence on the efficiency of the ECD process(es). From a systematic study on the impact of the filament heating current, filament bias voltage, and electron irradiation time on the intensities of precursor ions and various fragment ions, it was postulated that subsequent capture of electrons by the fragment ions, i.e., neutralization of the fragment ions, might be a significant event for limiting the intensity of the fragment ions.
Chan
T W Dominic
TW
Department of Chemistry, The Chinese University of Hong Kong, Shatin, NT, Hong Kong. twdchan@cuhk.edu.hk
Ip
W H Herman
WH
D000595:Amino Acid Sequence; D017357:Cyclotrons; D004583:Electrons; D005583:Fourier Analysis; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
D010455:Peptides