Accurate mass measurements using MALDI-TOF with delayed extraction.
Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry is now an essential tool in biopolymer analysis. Sensitivity and mass range are unsurpassed, but mass measurement accuracy and resolution have been limited. With delayed extraction and a reflecting analyzer, mass measurements using MALDI-TOF can be made with an accuracy of a few parts per million (ppm). It is possible to distinguish Lys from Gln in peptides, and to determine the elemental composition of smaller molecules (mass 100-500). In database searching strategies, a smaller mass window, resulting from an increase in mass accuracy, greatly decreases the number of possible candidates. Mass measurement accuracy with errors less than 5 ppm is demonstrated on a mixture of 12 peptides ranging in mass from ca. 900 to 3700 Da. Mass measurements on 13 peaks in an unseparated tryptic digest of myoglobin gave results with an overall average error less than 3.5 ppm, with a maximum error of 7 ppm.
Takach
E J
EJ
PerSeptive Biosystems, Inc., Framingham, Massachusetts 01701, USA.
Hines
W M
WM
Patterson
D H
DH
Juhasz
P
P
Falick
A M
AM
Vestal
M L
ML
Martin
S A
SA
D000324:Adrenocorticotropic Hormone / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D001059:Apoproteins / Q000032:analysis; D001920:Bradykinin / Q000031:analogs & derivatives / Q000032:analysis; D053497:Corticotropin-Like Intermediate Lobe Peptide; D005344:Fibrinopeptide A / Q000032:analysis; D006736:Horses; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009211:Myoglobin / Q000032:analysis; D009842:Oligopeptides / Q000032:analysis; D010446:Peptide Fragments / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D012015:Reference Standards; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D001059:Apoproteins; D009211:Myoglobin; D009842:Oligopeptides; D010446:Peptide Fragments; D011506:Proteins; C010857:apomyoglobin; C046722:bradykinin, des-Arg(1)-; D005344:Fibrinopeptide A; D053497:Corticotropin-Like Intermediate Lobe Peptide; D000324:Adrenocorticotropic Hormone; D001920:Bradykinin