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Title: Metabolite Spectral Accuracy on Orbitraps

Abstract

Orbitraps are high-resolution ion-trap mass spectrometers that are widely used in metabolomics. While the mass accuracy and resolving power of orbitraps have been extensively documented, their spectral accuracy, i.e., accuracy in measuring the abundances of isotopic peaks, remains less studied. In analyzing spectra of unlabeled metabolites, we discovered a systematic under representation of heavier natural isotopic species, especially for high molecular weight metabolites (~20% under-measurement of [M + 1]/[M + 0] ratio at m/z 600). We hypothesize that these discrepancies arise for metabolites far from the lower limit of the mass scan range, due to the weaker containment in the C-trap that results in suboptimal trajectories inside the Orbitrap analyzer. Consistent with this, spectral fidelity was restored by dividing the mass scan range (initially 75 m/z to 1000 m/z) into two scan events, one for lower molecular weight and the other for higher molecular weight metabolites. Having thus obtained accurate mass spectra at high resolution, we found that natural isotope correction for high-resolution labeling data requires more sophisticated algorithms than typically employed: the correction algorithm must take into account whether isotopologues with the same nominal mass are resolved. We present an algorithm and associated open-source code, named AccuCor, for thismore » purpose. Together, these improvements in instrument parameters and natural isotope correction enable more accurate measurement of metabolite labeling and thus metabolic flux.« less

Authors:
ORCiD logo [1];  [1];  [1]
  1. Princeton Univ., NJ (United States). Lewis Sigler Inst. for Integrative Genomics. Dept. of Chemistry
Publication Date:
Research Org.:
Princeton Univ., NJ (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Inst. of Health (NIH) (United States)
OSTI Identifier:
1534413
Grant/Contract Number:  
SC0012461; R01 CA163591; P30DK019525; CA211437
Resource Type:
Accepted Manuscript
Journal Name:
Analytical Chemistry
Additional Journal Information:
Journal Volume: 89; Journal Issue: 11; Journal ID: ISSN 0003-2700
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
isotopes; metabolism; labeling; ions

Citation Formats

Su, Xiaoyang, Lu, Wenyun, and Rabinowitz, Joshua D. Metabolite Spectral Accuracy on Orbitraps. United States: N. p., 2017. Web. doi:10.1021/acs.analchem.7b00396.
Su, Xiaoyang, Lu, Wenyun, & Rabinowitz, Joshua D. Metabolite Spectral Accuracy on Orbitraps. United States. doi:10.1021/acs.analchem.7b00396.
Su, Xiaoyang, Lu, Wenyun, and Rabinowitz, Joshua D. Thu . "Metabolite Spectral Accuracy on Orbitraps". United States. doi:10.1021/acs.analchem.7b00396. https://www.osti.gov/servlets/purl/1534413.
@article{osti_1534413,
title = {Metabolite Spectral Accuracy on Orbitraps},
author = {Su, Xiaoyang and Lu, Wenyun and Rabinowitz, Joshua D.},
abstractNote = {Orbitraps are high-resolution ion-trap mass spectrometers that are widely used in metabolomics. While the mass accuracy and resolving power of orbitraps have been extensively documented, their spectral accuracy, i.e., accuracy in measuring the abundances of isotopic peaks, remains less studied. In analyzing spectra of unlabeled metabolites, we discovered a systematic under representation of heavier natural isotopic species, especially for high molecular weight metabolites (~20% under-measurement of [M + 1]/[M + 0] ratio at m/z 600). We hypothesize that these discrepancies arise for metabolites far from the lower limit of the mass scan range, due to the weaker containment in the C-trap that results in suboptimal trajectories inside the Orbitrap analyzer. Consistent with this, spectral fidelity was restored by dividing the mass scan range (initially 75 m/z to 1000 m/z) into two scan events, one for lower molecular weight and the other for higher molecular weight metabolites. Having thus obtained accurate mass spectra at high resolution, we found that natural isotope correction for high-resolution labeling data requires more sophisticated algorithms than typically employed: the correction algorithm must take into account whether isotopologues with the same nominal mass are resolved. We present an algorithm and associated open-source code, named AccuCor, for this purpose. Together, these improvements in instrument parameters and natural isotope correction enable more accurate measurement of metabolite labeling and thus metabolic flux.},
doi = {10.1021/acs.analchem.7b00396},
journal = {Analytical Chemistry},
number = 11,
volume = 89,
place = {United States},
year = {2017},
month = {5}
}

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