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Automatic internal calibration in liquid chromatography/Fourier transform ion cyclotron resonance mass spectrometry of protein digests
Author(s) -
Palmblad Magnus,
Bindschedler Laurence V.,
Gibson Trevor M.,
Cramer Rainer
Publication year - 2006
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2707
Subject(s) - fourier transform ion cyclotron resonance , chemistry , mass spectrometry , chromatography , analytical chemistry (journal) , top down proteomics , protein mass spectrometry , tandem mass spectrometry , calibration , ion cyclotron resonance , selected ion monitoring , liquid chromatography–mass spectrometry , selected reaction monitoring , hybrid mass spectrometer , ion , gas chromatography–mass spectrometry , cyclotron , physics , organic chemistry , quantum mechanics
Accurately measured peptide masses can be used for large‐scale protein identification from bacterial whole‐cell digests as an alternative to tandem mass spectrometry (MS/MS) provided mass measurement errors of a few parts‐per‐million (ppm) are obtained. Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) routinely achieves such mass accuracy either with internal calibration or by regulating the charge in the analyzer cell. We have developed a novel and automated method for internal calibration of liquid chromatography (LC)/FTICR data from whole‐cell digests using peptides in the sample identified by concurrent MS/MS together with ambient polydimethylcyclosiloxanes as internal calibrants in the mass spectra. The method reduced mass measurement error from 4.3 ± 3.7 ppm to 0.3 ± 2.3 ppm in an E. coli LC/FTICR dataset of 1000 MS and MS/MS spectra and is applicable to all analyses of complex protein digests by FTICRMS. Copyright © 2006 John Wiley & Sons, Ltd.