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. 2019:2030:403-414.
doi: 10.1007/978-1-4939-9639-1_30.

Direct Analysis of Underivatized Amino Acids in Plant Extracts by LC-MS/MS (Improved Method)

Björn Thiele  1   2 , Michelle Hupert  3 , Beatrix Santiago-Schübel  3 , Marco Oldiges  4 , Diana Hofmann  5
Affiliations

Direct Analysis of Underivatized Amino Acids in Plant Extracts by LC-MS/MS (Improved Method)

Björn Thiele et al. Methods Mol Biol. 2019.

Abstract

In this chapter we describe a method for quantification of 20 proteinogenic amino acids by liquid chromatography-mass spectrometry which affords neither derivatization nor the use of organic solvents. Analysis of the underivatized amino acids is performed by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) in the positive ESI mode. Separation is achieved on a strong cation exchange (SCX) column (Luna 5 μ SCX 100 Å) with 5% acetic acid in water (A) and 75 mM ammonium acetate in water (B). Quantification is accomplished by use of d2-phenylalanine as internal standard achieving limits of detection of 5-50 nM. The method was successfully applied for the determination of proteinogenic amino acids in plant extracts.

Keywords: Amino acids; LC-MS/MS; Plant metabolism; Strong cation exchange column.

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