Stable isotope-resolved metabolomics (SIRM) is increasingly used among researchers for metabolic studies including amino acid metabolism. However, the classical GC- or HPLC-based methods for amino acid quantification do not meet the needs for multiplexed stable isotope-enriched analysis by ultrahigh-resolution Fourier transform mass spectrometry (UHR-FTMS). This is due to insufficient acquisition time during chromatographic separations and large dynamic range in concentrations of analytes, which compromises detection and quantification of the numerous metabolite isotopologues present in crude extracts. This chapter discusses a modified ethyl chloroformate derivatization method to enable rapid quantitative analysis of stable isotope-enriched amino acids using direct infusion ion introduction coupled with UHR-FTMS.
|Title of host publication||Methods in Molecular Biology|
|Number of pages||12|
|State||Published - 2019|
|Name||Methods in Molecular Biology|
Bibliographical noteFunding Information:
This work was supported in part by the National Institutes of Health grants [5R01ES22191-04, 3R01ES022191-04S1, 1U24DK097215-01A1, and P01 CA163223-01A1].
© Springer Science+Business Media, LLC, part of Springer Nature 2019.
- Amino acids
- Chloroformate derivatization
- Direct infusion nano-electrospray
- Stable isotope-resolved metabolomics
- Ultrahigh-resolution Fourier transform mass spectrometry
ASJC Scopus subject areas
- Molecular Biology