High-Resolution MRT-TOF Mass Spectrometry Enables Simultaneous Carbon and Nitrogen Tracing in Stable Isotope Resolved Metabolomics
4 affiliations
- 1School of Medicine
- 2University of Louisville
- 3UofL Health-Brown Cancer Center, University of Louisville
- 4Waters Corporation
Abstract
Stable isotope-resolved metabolomics (SIRM) using dual-isotope tracers, such as [U-13C5,15N2]glutamine, provides complementary insight into carbon and nitrogen metabolism and their integration across interconnected metabolic pathways. However, distinguishing closely spaced 13C- and 15N-containing isotopologues requires resolving power that conventional quadrupole time-of-flight mass spectrometers (QTOF MS) may not provide. In this study, we evaluated the performance of a multi-reflecting time-of-flight (MRT) mass spectrometer for dual-isotope SIRM using [U-13C5,15N2]glutamine-infused patient-derived tumor xenografts and compared its performance with that of a conventional SYNAPT™ XS QTOF Platform operated at a resolving power of approximately 15,000. A total of 113 metabolites were detected on both platforms, and dual-isotope labeling patterns were evaluated for 29 carbon- and nitrogen-containing metabolites. The Xevo™ MRT Mass Spectrometer, operating at a resolving power of approximately 100,000, resolved closely spaced 13C- and 15N isotopologues that remained unresolved on the SYNAPT™ XS Instrument under the acquisition conditions used. This separation enabled composition-specific isotopologue assignment, independent peak integration, natural-abundance correction, and isotopologue-specific fractional-enrichment analysis. The higher sensitivity of Xevo™ MRT Mass Spectrometer improved detection of low-abundance isotopologues, increasing isotopologue coverage when compared to SYNAPT™ XS MS Instrument. Together, these results demonstrate that MRT-TOF mass spectrometry provides a practical high-resolution platform for dual-isotope SIRM and enables simultaneous assessment of carbon incorporation and nitrogen-transfer reactions within a single experiment.
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BibTeX
@misc{silva2026highresolution,
title = {{High-Resolution MRT-TOF Mass Spectrometry Enables Simultaneous Carbon and Nitrogen Tracing in Stable Isotope Resolved Metabolomics}},
author = {Maleesha De Silva and Yoannis Imbert‑Fernandez and Brian F. Clem and Steven Lai and Pawel Lorkiewicz},
howpublished = {ChemRxiv},
year = {2026},
month = oct,
doi = {10.26434/chemrxiv.15010430/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15010430/v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - High-Resolution MRT-TOF Mass Spectrometry Enables Simultaneous Carbon and Nitrogen Tracing in Stable Isotope Resolved Metabolomics
AU - Maleesha De Silva
AU - Yoannis Imbert‑Fernandez
AU - Brian F. Clem
AU - Steven Lai
AU - Pawel Lorkiewicz
PY - 2026
DA - 2026/10/13
PB - ChemRxiv
DO - 10.26434/chemrxiv.15010430/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15010430/v1
AB - Stable isotope-resolved metabolomics (SIRM) using dual-isotope tracers, such as [U-13C5,15N2]glutamine, provides complementary insight into carbon and nitrogen metabolism and their integration across interconnected metabolic pathways. However, distinguishing closely spaced 13C- and 15N-containing isotopologues requires resolving power that conventional quadrupole time-of-flight mass spectrometers (QTOF MS) may not provide. In this study, we evaluated the performance of a multi-reflecting time-of-flight (MRT) mass spectrometer for dual-isotope SIRM using [U-13C5,15N2]glutamine-infused patient-derived tumor xenografts and compared its performance with that of a conventional SYNAPT™ XS QTOF Platform operated at a resolving power of approximately 15,000. A total of 113 metabolites were detected on both platforms, and dual-isotope labeling patterns were evaluated for 29 carbon- and nitrogen-containing metabolites. The Xevo™ MRT Mass Spectrometer, operating at a resolving power of approximately 100,000, resolved closely spaced 13C- and 15N isotopologues that remained unresolved on the SYNAPT™ XS Instrument under the acquisition conditions used. This separation enabled composition-specific isotopologue assignment, independent peak integration, natural-abundance correction, and isotopologue-specific fractional-enrichment analysis. The higher sensitivity of Xevo™ MRT Mass Spectrometer improved detection of low-abundance isotopologues, increasing isotopologue coverage when compared to SYNAPT™ XS MS Instrument. Together, these results demonstrate that MRT-TOF mass spectrometry provides a practical high-resolution platform for dual-isotope SIRM and enables simultaneous assessment of carbon incorporation and nitrogen-transfer reactions within a single experiment.
ER -