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A Strategy for Sensitive, Large Scale Quantitative Metabolomics
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Metabolite Spectral Accuracy on Orbitraps.

Xiaoyang Su1, Wenyun Lu1, Joshua D Rabinowitz1

  • 1Lewis Sigler Institute for Integrative Genomics and Department of Chemistry, Princeton University , Washington Road, Princeton, New Jersey 08544, United States.

Analytical Chemistry
|May 5, 2017
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Summary
This summary is machine-generated.

Orbitrap mass spectrometers often under-measure heavier isotopes in metabolites. Adjusting scan ranges and using the AccuCor algorithm improve spectral accuracy for better metabolic flux analysis.

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Area of Science:

  • Analytical Chemistry
  • Metabolomics
  • Mass Spectrometry

Background:

  • Orbitrap mass spectrometers are crucial for high-resolution metabolomics.
  • Spectral accuracy, particularly for isotopic peak abundances, is less understood than mass accuracy.
  • Existing methods may misrepresent natural isotope abundances in complex samples.

Purpose of the Study:

  • To investigate and address the systematic under-representation of heavier natural isotopes in Orbitrap mass spectrometry.
  • To improve the accuracy of metabolite labeling measurements and metabolic flux analysis.
  • To develop advanced algorithms for natural isotope correction in high-resolution metabolomics.

Main Methods:

  • Analysis of unlabeled metabolite spectra to identify isotopic under-representation.
  • Optimization of Orbitrap scan parameters by dividing the mass range into multiple scan events.
  • Development and validation of a novel natural isotope correction algorithm (AccuCor) accounting for isotopologue resolution.

Main Results:

  • A systematic under-measurement of heavier natural isotopes was observed, particularly for high molecular weight metabolites (up to 20% at m/z 600).
  • Dividing the mass scan range into separate events for lower and higher molecular weight metabolites restored spectral fidelity.
  • The AccuCor algorithm accurately corrects for natural isotopes, considering resolved isotopologues at high resolution.

Conclusions:

  • Optimizing Orbitrap scan parameters enhances spectral accuracy for metabolite analysis.
  • Advanced algorithms like AccuCor are necessary for precise natural isotope correction in high-resolution metabolomics.
  • These advancements lead to more accurate metabolite labeling quantification and metabolic flux determination.