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Open AccessArticle

Quantification of Glutamate and Aspartate by Ultra-High Performance Liquid Chromatography

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Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Liesel-Beckmann-Straße 1, 85354 Freising, Germany
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Chair of Plant Nutrition, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Emil-Ramann-Straße 2, 85350 Freising, Germany
*
Author to whom correspondence should be addressed.
Molecules 2018, 23(6), 1389; https://doi.org/10.3390/molecules23061389
Received: 22 May 2018 / Revised: 5 June 2018 / Accepted: 5 June 2018 / Published: 8 June 2018
(This article belongs to the Section Analytical Chemistry)
Glutamic and aspartic acid fulfil numerous functions in organisms. They are proteinogenic amino acids, they function as neurotransmitters, and glutamic acid links the citrate cycle with amino acid metabolism. In addition, glutamic acid is a precursor for many bioactive molecules like γ-aminobutyric acid (GABA). In tomatoes, glutamic acid accumulates in ripening fruits. Here we present a simple and rapid method for quantification of glutamate and aspartate in tomatoes. A cleared extract is prepared and 2-aminoadipic acid added as internal standard. Subsequently, the amino acids are derivatised with 2,4-dinitro-1-fluorobenzene under alkaline conditions. The derivatives are separated by ultra-high performance liquid chromatography using a phenyl-hexyl column and 50 mM N-methylmorpholine/acetate buffer pH 7.4 containing 12% acetonitrile as eluent and detected by UV absorption at 363 nm. The whole analysis time including separation and column equilibration takes less than 2.8 min with a flow rate of 1 mL/min and less than 1.6 min with a flow rate of 2 mL/min, making this method suitable for high-throughput applications. The method shows excellent reproducibility with intra- and inter-day SDs of approximately 4% for both aspartic and glutamic acid. Using this method we show that the glutamate/aspartate ratio changes significantly during fruit ripening. View Full-Text
Keywords: aspartic acid; 2,4-dinitro-1-fluorobenzene; glutamic acid; ripening; tomato; ultra-high performance liquid chromatography aspartic acid; 2,4-dinitro-1-fluorobenzene; glutamic acid; ripening; tomato; ultra-high performance liquid chromatography
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MDPI and ACS Style

Agius, C.; Von Tucher, S.; Poppenberger, B.; Rozhon, W. Quantification of Glutamate and Aspartate by Ultra-High Performance Liquid Chromatography. Molecules 2018, 23, 1389.

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