Improved segmented-scan spectral stitching for stable isotope resolved metabolomics (SIRM) by ultra-high-resolution Fourier transform mass spectrometry

Anal Chim Acta. 2019 Nov 8:1080:104-115. doi: 10.1016/j.aca.2019.06.019. Epub 2019 Jun 11.

Abstract

We have implemented a linear ion trap (LIT)-based SIM-stitching method for ultra-high-resolution Fourier transform mass spectrometry (FTMS) that increases the S/N over a wide m/z range compared to non-segmented wide full-scan (WFS) spectra. Here we described an improved segmented spectral scan stitching method that was based on quadrupole mass filter (QMF)-SIM, which overcame previous limitations of ion signal loss in LIT. This allowed for accurate representation of isotopologue distributions, both at natural abundance and in stable isotope-resolved metabolomics (SIRM)-based experiments. We also introduced a new spectral binning method that provided more precise and resolution-independent bins for irreversibly noise-suppressed FTMS spectra. We demonstrated a substantial improvement in S/N and sensitivity (typically > 10-fold) for 13C labeled lipid extracts of human macrophages grown as three-dimensional (3D) cell culture, with detection of an increased number of 13C isotopologue ions. The method also enabled analysis of extracts from very limited biological samples.

Keywords: Spectral binning; Spectral stitching; Stable isotope-resolved metabolomics (SIRM); Ultra-high-resolution Fourier transform mass spectrometry.

MeSH terms

  • Carbon Isotopes / chemistry
  • Fourier Analysis
  • Glucose / chemistry
  • Glucose / metabolism
  • Humans
  • Isotope Labeling
  • Lipids / analysis*
  • Macrophages / chemistry*
  • Macrophages / metabolism
  • Mass Spectrometry / methods
  • Metabolomics / methods
  • Spheroids, Cellular / chemistry
  • Spheroids, Cellular / metabolism

Substances

  • Carbon Isotopes
  • Lipids
  • Carbon-13
  • Glucose