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| Home > Publications database > Analysis of amino acids without derivatization in barley extracts by LC-MS-MS |
| Journal Article | PreJuSER-62860 |
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2008
Springer
Berlin
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Please use a persistent id in citations: doi:10.1007/s00216-008-2167-9
Abstract: A method has been developed for quantification of 20 amino acids as well as 13 (15)N-labeled amino acids in barley plants. The amino acids were extracted from plant tissues using aqueous HCl-ethanol and directly analyzed without further purification. Analysis of the underivatized amino acids was performed by liquid chromatography (LC)-electrospray ionization (ESI) tandem mass spectrometry (MS-MS) in the positive ESI mode. Separation was achieved on a strong cation exchange column (Luna 5micro SCX 100A) with 30 mM ammonium acetate in water (solvent A) and 5% acetic acid in water (solvent B). Quantification was accomplished using d (2)-Phe as an internal standard. Calibration curves were linear over the range 0.5-50 microM, and limits of detection were estimated to be 0.1-3.0 microM. The mass-spectrometric technique was employed to study the regulation of amino acid levels in barley plants grown at 15 degrees C uniform root temperature (RT) and 20-10 degrees C vertical RT gradient (RTG). The LC-MS-MS results demonstrated enhanced concentration of free amino acids in shoots at 20-10 degrees C RTG, while total free amino acid concentration in roots was similarly low for both RT treatments. (15)NO(3) (-) labeling experiments showed lower (15)N/(14)N ratios for Glu, Ser, Ala and Val in plants grown at 20-10 degrees C RTG compared with those grown at 15 degrees C RT.
Keyword(s): Amino Acids: analysis (MeSH) ; Calibration (MeSH) ; Cations: chemistry (MeSH) ; Chromatography, Ion Exchange: methods (MeSH) ; Hordeum: chemistry (MeSH) ; Hordeum: metabolism (MeSH) ; Nitrogen Isotopes (MeSH) ; Plant Extracts: analysis (MeSH) ; Plant Leaves: chemistry (MeSH) ; Plant Leaves: metabolism (MeSH) ; Plant Roots: chemistry (MeSH) ; Plant Roots: metabolism (MeSH) ; Plant Shoots: chemistry (MeSH) ; Plant Shoots: metabolism (MeSH) ; Reproducibility of Results (MeSH) ; Spectrometry, Mass, Electrospray Ionization: methods (MeSH) ; Tandem Mass Spectrometry: methods (MeSH) ; Amino Acids ; Cations ; Nitrogen Isotopes ; Plant Extracts ; J ; liquid chromatography-mass spectrometry (auto) ; amino acids (auto) ; strong cation exchange column (auto) ; plant metabolism (auto)
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