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. 2019:2030:57-68.
doi: 10.1007/978-1-4939-9639-1_6.

Quantification of Isotopologues of Amino Acids by Multiplexed Stable Isotope-Resolved Metabolomics Using Ultrahigh-Resolution Mass Spectrometry Coupled with Direct Infusion

Ye Yang  1 , Teresa W-M Fan  2   3 , Andrew N Lane  4   5 , Richard M Higashi  4   5
Affiliations

Quantification of Isotopologues of Amino Acids by Multiplexed Stable Isotope-Resolved Metabolomics Using Ultrahigh-Resolution Mass Spectrometry Coupled with Direct Infusion

Ye Yang et al. Methods Mol Biol. 2019.

Abstract

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.

Keywords: Amino acids; Chloroformate derivatization; Direct infusion nano-electrospray; Stable isotope-resolved metabolomics; Ultrahigh-resolution Fourier transform mass spectrometry.

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Figures

Figure 1.
Figure 1.
Reaction scheme of ECF derivatization of amino acids [35].
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