| Evaluation and optimization of electron capture dissociation efficiency in fourier transform ion cyclotron resonance mass spectrometry. | |
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MedLine Citation:
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PMID: 15914017 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Electron capture dissociation (ECD) efficiency has typically been lower than for other dissociation techniques. Here we characterize experimental factors that limit ECD and seek to improve its efficiency. Efficiency of precursor to product ion conversion was measured for a range of peptide (approximately 15% efficiency) and protein (approximately 33% efficiency) ions of differing sizes and charge states. Conversion of precursor ions to products depends on electron irradiation period and maximizes at approximately 5-30 ms. The optimal irradiation period scales inversely with charge state. We demonstrate that reflection of electrons through the ICR cell is more efficient and robust than a single pass, because electrons can cool to the optimal energy for capture, which allows for a wide range of initial electron energy. Further, efficient ECD with reflected electrons requires only a short (approximately 500 micros) irradiation period followed by an appropriate delay for cooling and interaction. Reflection of the electron beam results in electrons trapped in or near the ICR cell and thus requires a brief (approximately 50 micros) purge for successful mass spectral acquisition. Further electron irradiation of refractory precursor ions did not result in further dissociation. Possibly the ion cloud and electron beam are misaligned radially, or the electron beam diameter may be smaller than that of the ion cloud such that remaining precursor ions do not overlap with the electron beam. Several ion manipulation techniques and use of a large, movable dispenser cathode reduce the possibility that misalignment of the ion and electron beams limits ECD efficiency. |
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Authors:
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Melinda A McFarland; Michael J Chalmers; John P Quinn; Christopher L Hendrickson; Alan G Marshall |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Journal of the American Society for Mass Spectrometry Volume: 16 ISSN: 1044-0305 ISO Abbreviation: J. Am. Soc. Mass Spectrom. Publication Date: 2005 Jul |
Date Detail:
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Created Date: 2005-06-24 Completed Date: 2005-08-30 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9010412 Medline TA: J Am Soc Mass Spectrom Country: United States |
Other Details:
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Languages: eng Pagination: 1060-6 Citation Subset: IM |
Affiliation:
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Ion Cyclotron Resonance Program, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida 32310-4005, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Cyclotrons* Glutamic Acid / chemistry Melitten / chemistry Peptides / chemistry* Spectrometry, Mass, Electrospray Ionization / methods Spectroscopy, Fourier Transform Infrared / methods* Substance P / chemistry Ubiquitin / chemistry |
| Chemical | |
Reg. No./Substance:
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0/Peptides; 0/Ubiquitin; 20449-79-0/Melitten; 33507-63-0/Substance P; 56-86-0/Glutamic Acid |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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