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Fast quantification of amino acids by microchip electrophoresis–mass spectrometry

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Abstract

A fast microchip electrophoresis–nano-electrospray ionization-mass spectrometric method (MCE-nanoESI-MS) was developed for analysis of amino acids in biological samples. A glass/poly(dimethylsiloxane) hybrid microchip with a monolithic nanoESI emitter was used in the platform. The proposed MCE-nanoESI-MS analytical method showed high separation efficiency for amino acids. Baseline separation of an amino acid mixture containing Lys, Arg, Val, Tyr, and Glu was completed within 120 s with theoretical plate numbers of >7,500. The method was applied to study cellular release of excitatory amino acids (i.e., aspartic acid (Asp) and glutamic acid (Glu)) under chemical stimulations. Linear calibration curves were obtained for both Asp and Glu in a concentration range from 1.00 to 150.0 μM. Limits of detection were found to be 0.37 μM for Asp and 0.33 μM for Glu (S/N = 3). Assay repeatability (relative standard deviation, n = 6) was 4.2 and 4.5 %, for Asp and Glu at 5.0 μM, respectively. In the study of cellular release, PC-12 nerve cells were incubated with alcohol at various concentrations for 1 h. Both extra- and intracellular levels of Asp and Glu were measured by the proposed method. The results clearly indicated that ethanol promoted the release of both Asp and Glu from the cells.

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Acknowledgments

Financial support from US NIH (GM089557 to YML and G12MD007581-15 to PBT) is gratefully acknowledged.

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Authors and Affiliations

  1. College of Chemistry, Sichuan University, Chengdu, 610065, China

    Xiangtang Li & Dan Xiao

  2. Department of Biology, Jackson State University, 1400 Lynch St., Jackson, MS, 39217, USA

    Talia Sanders & Paul B. Tchounwou

  3. Department of Chemistry and Biochemistry, Jackson State University, 1400 Lynch St., Jackson, MS, 39217, USA

    Xiangtang Li & Yi-Ming Liu

Authors
  1. Xiangtang Li
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  2. Dan Xiao
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  3. Talia Sanders
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  4. Paul B. Tchounwou
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  5. Yi-Ming Liu
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Corresponding authors

Correspondence to Dan Xiao or Yi-Ming Liu.

Additional information

Published in the topical collection Amino Acid Analysis with guest editor Toshimasa Toyo'oka.

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Li, X., Xiao, D., Sanders, T. et al. Fast quantification of amino acids by microchip electrophoresis–mass spectrometry. Anal Bioanal Chem 405, 8131–8136 (2013). https://doi.org/10.1007/s00216-013-7260-z

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  • DOI: https://doi.org/10.1007/s00216-013-7260-z

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