Microscale analysis of amino acids using gas chromatography-mass spectrometry after methyl chloroformate derivatization

Wen Ping Chen, Xiao Yuan Yang, Adrian D. Hegeman, William M. Gray, Jerry D. Cohen

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46 Scopus citations


To conduct studies of stable isotope incorporation and dilution in growing plants, a rapid microscale method for determination of amino acid profiles from minute amounts of plant samples was developed. The method involves solid-phase ion exchange followed by derivatization and analysis by gas chromatography-mass spectrometry (GC-MS). The procedure allowed the eluent to be derivatized directly with methyl chloroformate without sample lyophilization or other evaporation procedures. Sample extraction and derivatization required only ca. 30min and quantification of the 19 amino acids eluted from the cation exchange solid-phase extraction step from a single cotyledon (0.4mg fresh weight) or three etiolated 7-day-old Arabidopsis seedlings (0.1mg fresh weight) was easily accomplished in the selected ion monitoring mode. This method was especially useful for monitoring mass isotopic distribution of amino acids as illustrated by Arabidopsis seedlings that had been labeled with deuterium oxide and 15N salts. Sample preparation was facile, rapid, economical, and the method is easily modified for integration into robotic systems for analysis with large numbers of samples.

Original languageEnglish (US)
Pages (from-to)2199-2208
Number of pages10
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Issue number24
StatePublished - Aug 2010

Bibliographical note

Funding Information:
We thank Dr. Michael MacCoss at the University of Washington for sharing IDCal for calculation of theoretical amino acid isotopomer distributions. This work was supported by the U.S. National Science Foundation, Plant Genome Program, grants DBI-0606666 and IOS-0923960, and by funds from the Gordon and Margaret Bailey Endowment for Environmental Horticulture.


  • Alkyl chloroformate
  • Arabidopsis
  • GC-MS
  • Metabolite profiling
  • SPE column
  • Stable isotope


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