Matrix-assisted laser desorption using a fast-atom bombardment ion source and a magnetic mass spectrometer.
A conventional fast-atom bombardment (FAB) ion source was used to achieve matrix-assisted laser desorption (MALD) in a high-mass, double-focusing, magnetic mass spectrometer. The pulsed ion signals generated by irradiation of a mixture of sample and matrix (2,5-dihydroxybenzoic acid) with either a XeF excimer laser (353 nm) or a nitrogen laser (337 nm) were recorded with a focal-plane detector. A resolution (full-width at half maximum) of 4500 was achieved at m/z 1347.7 (the peptide substance P), 2500 for CsI cluster ions at m/z 10,005.7, and 1250 for the isotope cluster of the small protein cytochrome c (horse) [M+H]+ = m/z 12,360 (average). Sensitivity is demonstrated with 11 fmol of substance P. A survey scan is taken to locate the m/z of the sample molecular ion. The segment that contains the sample can then be integrated for a longer time to produce a better signal-to-noise ratio. In addition to higher sensitivity and lower matrix interference, the advantage of MALD over FAB is the former's lower susceptibility to the presence of salts, and competition between hydrophobic and hydrophilic components of a mixture. This feature is demonstrated by the complete MALD spectrum of a crude partial tryptic digest of sperm-whale apomyoglobin, containing 24 peptides, representing the entire sequence of this protein.
Annan
R S
RS
Department of Chemistry, Massachusetts Institute of Technology, Cambridge 02139.
Köchling
H J
HJ
Hill
J A
JA
Biemann
K
K
D000818:Animals; D001059:Apoproteins / Q000032:analysis; D002417:Cattle; D003574:Cytochrome c Group / Q000032:analysis; D006736:Horses; D007834:Lasers; D008280:Magnetics; D013058:Mass Spectrometry / Q000295:instrumentation*; D009211:Myoglobin / Q000032:analysis; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013373:Substance P / Q000032:analysis; D014452:Ubiquitins / Q000032:analysis; D014907:Whales
D001059:Apoproteins; D003574:Cytochrome c Group; D009211:Myoglobin; D014452:Ubiquitins; C010857:apomyoglobin; D013373:Substance P