Proposing a validation scheme for ¹³C metabolite tracer studies in high-resolution mass spectrometry.

ABSTRACT: ¹³C metabolite tracer and metabolic flux analyses require upfront experimental planning and validation tools. Here, we present a validation scheme including a comparison of different LC methods that allow for customization of analytical strategies for tracer studies with regard to the targeted metabolites. As the measurement of significant changes in labeling patterns depends on the spectral accuracy, we investigate this aspect comprehensively for high-resolution orbitrap mass spectrometry combined with reversed-phase chromatography, hydrophilic interaction liquid chromatography, or anion-exchange chromatography. Moreover, we propose a quality control protocol based on (1) a metabolite containing selenium to assess the instrument performance and on (2) in vivo synthesized isotop

SUBMITTER: Schwaiger-Haber M 

PROVIDER: S-EPMC6526147 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature