Profiling of Endogenous Metabolites Using TOF LC/MS with Ion-Pair Reverse Phase Chromatography

A high performance ion-pair reverse-phase (IP-RP) chromatographic method
has been developed for use with a time-of-flight (TOF) mass spectrometer (MS).
This negative ionization method provides comprehensive coverage of many
endogenous metabolite classes including amino acids, organic acids, sugars and
sugar phosphates, nucleosides and nucleotides, energy and redox metabolites, and
coenzyme A derivatives. Optimization of mobile phase pH and the LC gradient was
critical in obtaining good chromatographic separation of isomers such as citric
acid and isocitric acid, and impacted peak shape and optimal signal response.
The IP-RP LC/TOF MS method was evaluated with 19 representative metabolite
standards covering the entire chromatographic region. All metabolites showed
good retention time reproducibility

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