title: Analysis of a tryptic digest of pig hemoglobin using ESI-FAIMS-MS.
abstract: The continuous gas-phase ion separation and atmospheric pressure focusing properties of high-field asymmetric waveform ion mobility spectrometry (FAIMS) offer significant advantages for the mass spectrometric analysis of tryptic digests of proteins. In this study, tryptic peptides of pig hemoglobin were examined by ESI-FAIMS-MS using a newly designed FAIMS device. The new, hemispherical geometry of the inner electrode served to deliver the ions, via the gas flows, to the center axis of the FAIMS analyzer, improving the sensitivity relative to previous prototypes. Mass spectra collected using this new FAIMS showed significantly less chemical background noise than conventional ESI-MS, while maintaining approximately the same absolute sensitivity as that observed with ESI-MS. As a consequence of the ion separation in FAIMS, the identification of the tryptic fragments was simplified and some peptides, such as the triply protonated WAGVANALAHK3+, that were obscured by the intense background of ESI-MS, were readily detected using ESI-FAIMS-MS. In addition, the FAIMS device was shown to separate isobaric ions at m/z 532.4. Correlations between CV and mass-to-charge ratio, as well as CV and ionic collision cross section, were evaluated for 38 peptide ions identified in the tryptic digest. The correlation between the CV of the peptide and the mass-to-charge ratio is very poor, indicating good orthogonality between the separation by FAIMS and the separation by mass spectrometry.
authors.affiliation: Institute for National Measurement Standards, National Research Council of Canada, Ottawa. roger.guevremont@nrc.ca
authors.forename: R
authors.initials: R
authors.lastname: Guevremont
authors.forename: D A
authors.initials: DA
authors.lastname: Barnett
authors.forename: R W
authors.initials: RW
authors.lastname: Purves
authors.forename: J
authors.initials: J
authors.lastname: Vandermey
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006454:Hemoglobins / Q000737:chemistry*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D013552:Swine; D014357:Trypsin / Q000494:pharmacology
chemical_list: D006454:Hemoglobins; D014357:Trypsin