This title appears in the Scientific Report :
2008
Please use the identifier:
http://dx.doi.org/10.1007/s00216-008-2167-9 in citations.
Analysis of amino acids without derivatization in barley extracts by LC-MS-MS
Analysis of amino acids without derivatization in barley extracts by LC-MS-MS
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 per...
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Personal Name(s): | Thiele, B. |
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Füllner, K. / Stein, N. / Oldiges, M. / Kuhn, A. J. / Hofmann, D. | |
Contributing Institute: |
Zentralabteilung für Chemische Analysen; ZCH Biotechnologie 2; IBT-2 Phytosphäre; ICG-3 |
Published in: | Analytical and bioanalytical chemistry, 391 (2008) S. 2663 - 2672 |
Imprint: |
Berlin
Springer
2008
|
Physical Description: |
2663 - 2672 |
PubMed ID: |
18506428 |
DOI: |
10.1007/s00216-008-2167-9 |
Document Type: |
Journal Article |
Research Program: |
Terrestrische Umwelt |
Series Title: |
Analytical and Bioanalytical Chemistry
391 |
Subject (ZB): | |
Publikationsportal JuSER |
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. |