Dataset Information

An optimization method for untargeted MS-based isotopic tracing investigations of metabolism

ABSTRACT:

Introduction

Stable isotope tracer studies are increasingly applied to explore metabolism from the detailed analysis of tracer incorporation into metabolites. Untargeted LC/MS approaches have recently emerged and provide potent methods for expanding the dimension and complexity of the metabolic networks that can be investigated. A number of software tools have been developed to process the highly complex MS data collected in such studies; however, a method to optimize the extraction of valuable isotopic data is lacking.

Objectives

To develop and validate a method to optimize automated data processing for untargeted MS-based isotopic tracing investigations of metabolism.

Methods

The method is based on the application of a suitable reference material to rationally perform parameter optimization throughout the complete data processing workflow. It was applied in the context of 13C-labelling experiments and with two different software, namely geoRge and X13CMS. It was illustrated with the study of a E. coli mutant impaired for central metabolism.

Results

The optimization methodology provided significant gain in the number and quality of extracted isotopic data, independently of the software considered. Pascal triangle samples are well suited for such purpose since they allow both the identification of analytical issues and optimization of data processing at the same time.

Conclusion

The proposed method maximizes the biological value of untargeted MS-based isotopic tracing investigations by revealing the full metabolic information that is encoded in the labelling patterns of metabolites.

Supplementary Information

The online version contains supplementary material available at 10.1007/s11306-022-01897-5.

SUBMITTER: Butin N

PROVIDER: S-EPMC9205802 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature

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