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.
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Acknowledgment
The authors like to thank Bernd Kastenholz for his assistance in the optimization of the leaf extraction procedure.
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Thiele, B., Hupert, M., Santiago-Schübel, B., Oldiges, M., Hofmann, D. (2019). Direct Analysis of Underivatized Amino Acids in Plant Extracts by LC-MS/MS (Improved Method). In: Alterman, M. (eds) Amino Acid Analysis. Methods in Molecular Biology, vol 2030. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9639-1_30
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DOI: https://doi.org/10.1007/978-1-4939-9639-1_30
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