Matrix-assisted laser desorption ionization is used to obtain Fourier-transform ion cyclotron resonance mass spectra of model peptides (e.g., gramicidin S, angiotensin I, renin substrate, melittin, and bovine insulin). Matrix-assisted laser desorption ionization yields ions having appreciable kinetic energies. Two methods for trapping the high kinetic energy ions are described: (') the ion signal for [M + H]+ ions is shown to increase with increasing trapping voltages, and (ii) collisional relaxation is used for the detection of [M + H]+ ions of bovine insulin.Ionization methods such as californium plasma desorption (1), matrix-assisted laser desorption (MALD) (2), and electrospray ionization (ESI) (3) have greatly expanded the role of mass spectrometry of high mass biomolecules. Biomolecules having molecular masses >100,000 Da can be ionized to yield a single-charged (e.g., [M + H]+) or multiple-charged (e.g., [M + nH]n+) ion. Although most of the MALD and ESI work has utilized time-of-flight, quadrupole mass spectrometers and/or quadrupole ion traps, MALD (4) and ESI are adaptable to Fourier-transform ion cyclotron resonance (FT-ICR). The advantage of FTICR (and the quadrupole ion trap) is that tandem mass spectrometry experiments [e.g., collision-induced dissociation and photodissociation (5)] and ion molecule reactions can be implemented for structural characterization. The best high mass performance [-60,000 mass resolution at m/z 6000 (6)] of FTICR has been achieved with laser desorption ionization of nonpolar polymers. McLafferty and coworkers (7) have made considerable progress with the development of ESI for FTICR, and a highsensitivity ESI/FTICR system was recently described.* Several groups (9, 10) have suggested that the performance of FTICR with biological molecules (samples that have relatively low ionization probabilities) and molecular masses greater than -2500 Da may be adversely affected by inefficient trapping of the ions or inherent limitations of the ion detection. For example, the mass resolution for samples with molecular masses greater than -2500 Da is far less than the theoretically predicted values (11). On the basis of our experience with desorption ionization and high mass FTICR, we suggested that the major problem with high-resolution FTICR at high mass is due to the high kinetic energies of desorbed ions (9). We have recently evaluated two experimental methods for improving the trapping efficiency of high mass ions: (i) the effects of increasing the electric trapping field and (ii) the effects of collisional relaxation of the laser-desorbed species prior to trapping the ions in the ion cyclotron resonance (ICR) cell. In this article we present the results from both experimental methods. The results show that increasing the trapping potentials improves the ion trapping efficiency, but detection of [M + H]+ ions of bovine insulin (m/z = 5734) requires collisional relaxation of ions prior to their introduction into the ion cell.
MATERIALS AND METHODSFTICR Mass Spectrometer. The...