Cargando…

Axial spatial distribution focusing: improving MALDI-TOF/RTOF mass spectrometric performance for high-energy collision-induced dissociation of biomolecules

RATIONALE: For the last two decades, curved field reflectron technology has been used in matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometers, assisting in the generation of post-source-decay (PSD) or collision-induced dissociation (CID) without decelerating prec...

Descripción completa

Detalles Bibliográficos
Autores principales: Belgacem, O, Pittenauer, E, Openshaw, M E, Hart, P J, Bowdler, A, Allmaier, G
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Ltd 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4699192/
https://www.ncbi.nlm.nih.gov/pubmed/26754126
http://dx.doi.org/10.1002/rcm.7458
_version_ 1782408152952602624
author Belgacem, O
Pittenauer, E
Openshaw, M E
Hart, P J
Bowdler, A
Allmaier, G
author_facet Belgacem, O
Pittenauer, E
Openshaw, M E
Hart, P J
Bowdler, A
Allmaier, G
author_sort Belgacem, O
collection PubMed
description RATIONALE: For the last two decades, curved field reflectron technology has been used in matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometers, assisting in the generation of post-source-decay (PSD) or collision-induced dissociation (CID) without decelerating precursor ions, producing true high-energy CID spectra. The result was the generation of product ion mass spectra with product ions typical of high-energy (10 keV and beyond) collision processes. The disadvantage of this approach was the lack of resolution in CID spectra resulting from the excess laser energy deposition used to generate those MS/MS spectra. The work presented in this study overcomes this limitation and includes comprehensive examples of high-energy and high-resolution CID MALDI-MS/MS spectra of biomolecules. METHODS: The devices used in this study are TOF/RTOF instruments equipped with a high-vacuum MALDI ion source. High-resolution and high-energy CID spectra result from the use of axial spatial distribution focusing (ASDF) in combination with curved field reflectron technology. RESULTS: A CID spectrum of the P(14)R(1) peptide exhibits product ion resolution in excess of 10,000 (FWHM) but at the same time yields typical high-energy product ions such as w- and [y–2]-type ion series. High-energy CID spectra of lipids, exemplified by a glycerophospholipid and triglyceride, demonstrate C–C backbone fragmentation elucidating the presence of a hydroxyl group in addition to double-bond positioning. A complex high mannose carbohydrate (Man)(8)(GlcNAc)(2) was also studied at 20 keV collision energy and revealed further high-energy product ions with very high resolution, allowing unambiguous detection and characterization of cross-ring cleavage-related ions. CONCLUSIONS: This is the first comprehensive study using a MALDI-TOF/RTOF instrument equipped with a curved field reflectron and an ASDF device prior to the reflectron. © 2015 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.
format Online
Article
Text
id pubmed-4699192
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher John Wiley & Sons, Ltd
record_format MEDLINE/PubMed
spelling pubmed-46991922016-01-08 Axial spatial distribution focusing: improving MALDI-TOF/RTOF mass spectrometric performance for high-energy collision-induced dissociation of biomolecules Belgacem, O Pittenauer, E Openshaw, M E Hart, P J Bowdler, A Allmaier, G Rapid Commun Mass Spectrom Research Articles RATIONALE: For the last two decades, curved field reflectron technology has been used in matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometers, assisting in the generation of post-source-decay (PSD) or collision-induced dissociation (CID) without decelerating precursor ions, producing true high-energy CID spectra. The result was the generation of product ion mass spectra with product ions typical of high-energy (10 keV and beyond) collision processes. The disadvantage of this approach was the lack of resolution in CID spectra resulting from the excess laser energy deposition used to generate those MS/MS spectra. The work presented in this study overcomes this limitation and includes comprehensive examples of high-energy and high-resolution CID MALDI-MS/MS spectra of biomolecules. METHODS: The devices used in this study are TOF/RTOF instruments equipped with a high-vacuum MALDI ion source. High-resolution and high-energy CID spectra result from the use of axial spatial distribution focusing (ASDF) in combination with curved field reflectron technology. RESULTS: A CID spectrum of the P(14)R(1) peptide exhibits product ion resolution in excess of 10,000 (FWHM) but at the same time yields typical high-energy product ions such as w- and [y–2]-type ion series. High-energy CID spectra of lipids, exemplified by a glycerophospholipid and triglyceride, demonstrate C–C backbone fragmentation elucidating the presence of a hydroxyl group in addition to double-bond positioning. A complex high mannose carbohydrate (Man)(8)(GlcNAc)(2) was also studied at 20 keV collision energy and revealed further high-energy product ions with very high resolution, allowing unambiguous detection and characterization of cross-ring cleavage-related ions. CONCLUSIONS: This is the first comprehensive study using a MALDI-TOF/RTOF instrument equipped with a curved field reflectron and an ASDF device prior to the reflectron. © 2015 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd. John Wiley & Sons, Ltd 2016-02-15 2015-12-29 /pmc/articles/PMC4699192/ /pubmed/26754126 http://dx.doi.org/10.1002/rcm.7458 Text en © 2015 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Belgacem, O
Pittenauer, E
Openshaw, M E
Hart, P J
Bowdler, A
Allmaier, G
Axial spatial distribution focusing: improving MALDI-TOF/RTOF mass spectrometric performance for high-energy collision-induced dissociation of biomolecules
title Axial spatial distribution focusing: improving MALDI-TOF/RTOF mass spectrometric performance for high-energy collision-induced dissociation of biomolecules
title_full Axial spatial distribution focusing: improving MALDI-TOF/RTOF mass spectrometric performance for high-energy collision-induced dissociation of biomolecules
title_fullStr Axial spatial distribution focusing: improving MALDI-TOF/RTOF mass spectrometric performance for high-energy collision-induced dissociation of biomolecules
title_full_unstemmed Axial spatial distribution focusing: improving MALDI-TOF/RTOF mass spectrometric performance for high-energy collision-induced dissociation of biomolecules
title_short Axial spatial distribution focusing: improving MALDI-TOF/RTOF mass spectrometric performance for high-energy collision-induced dissociation of biomolecules
title_sort axial spatial distribution focusing: improving maldi-tof/rtof mass spectrometric performance for high-energy collision-induced dissociation of biomolecules
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4699192/
https://www.ncbi.nlm.nih.gov/pubmed/26754126
http://dx.doi.org/10.1002/rcm.7458
work_keys_str_mv AT belgacemo axialspatialdistributionfocusingimprovingmalditofrtofmassspectrometricperformanceforhighenergycollisioninduceddissociationofbiomolecules
AT pittenauere axialspatialdistributionfocusingimprovingmalditofrtofmassspectrometricperformanceforhighenergycollisioninduceddissociationofbiomolecules
AT openshawme axialspatialdistributionfocusingimprovingmalditofrtofmassspectrometricperformanceforhighenergycollisioninduceddissociationofbiomolecules
AT hartpj axialspatialdistributionfocusingimprovingmalditofrtofmassspectrometricperformanceforhighenergycollisioninduceddissociationofbiomolecules
AT bowdlera axialspatialdistributionfocusingimprovingmalditofrtofmassspectrometricperformanceforhighenergycollisioninduceddissociationofbiomolecules
AT allmaierg axialspatialdistributionfocusingimprovingmalditofrtofmassspectrometricperformanceforhighenergycollisioninduceddissociationofbiomolecules