Simultaneous determination of 24 free amino acids in MGC803 cells by hydrophilic interaction liquid chromatography with tandem mass spectrometry.

Abstract

Amino acids play key roles in cellular protein biosynthesis and energy metabolism pathways. In this study, a simple, rapid and sensitive method was developed for the simultaneous determination of 24 free amino acids in cell samples using hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry (HILIC-MS/MS). Cell samples were deproteinized with methanol/H2O (80:20, v/v) without intricate derivatization process. The analytes were separated on a Waters BEH Amide column (2.1 mm × 100 mm, 1.7 μm), and accomplished within 5 min at a flow rate of 0.2 mL/min. The good linearity was obtained for all analytes (r2 > 0.99) with the limits of quantification from 0.1 to 25 ng/mL. The intra- and inter-day precision ranged from 0.35 to 10.36% and from 2.22 to 9.93%, respectively. The recoveries of most analytes were between 80% and 120% with RSD less than 10.0%. The developed method was then applied to the direct analysis of 24 underivatized amino acids in human gastric cancer cell line MGC803 treated with the antitumor candidate drug J3, and significant differences in the concentration levels of amino acids were also assessed.

• Authors: Juan Li, Qing-Li Wang, Ying Liu, Yu Ke, Qi-Qi Fan, Piao Zhou, Meng-Chen An, Hong-Min Liu
• Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (J Chromatogr B Analyt Technol Biomed Life Sci) Vol. 1132 Pg. 121792 (Nov 15 2019) ISSN: 1873-376X [Electronic] Netherlands
• PMID: 31677481 (Publication Type: Journal Article)
• Copyright: Copyright © 2019 Elsevier B.V. All rights reserved.

Chemical References

• Amino Acids

Topics

• Amino Acids (analysis)
• Cell Line, Tumor
• Chromatography, Liquid (methods)
• Humans
• Hydrophobic and Hydrophilic Interactions
• Linear Models
• Reproducibility of Results
• Sensitivity and Specificity
• Tandem Mass Spectrometry (methods)