Rapid quantification of glutaminase 2 (GLS2)-related metabolites by HILIC-MS/MS

Guan-Yuan Chen; Hsi-Chun Chao; Hsiao-Wei Liao; I-Lin Tsai; Ching-Hua Kuo

doi:10.1016/j.ab.2017.10.002

Rapid quantification of glutaminase 2 (GLS2)-related metabolites by HILIC-MS/MS

Anal Biochem. 2017 Dec 15:539:39-44. doi: 10.1016/j.ab.2017.10.002. Epub 2017 Oct 6.

Authors

Guan-Yuan Chen¹, Hsi-Chun Chao¹, Hsiao-Wei Liao¹, I-Lin Tsai², Ching-Hua Kuo³

Affiliations

¹ School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan; The Metabolomics Core Laboratory, Center of Genomic Medicine, National Taiwan University, Taipei, Taiwan.
² Department of Biochemistry and Molecular Cell Biology, College of Medicine, Taipei Medical University, Taipei, Taiwan; Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
³ School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan; The Metabolomics Core Laboratory, Center of Genomic Medicine, National Taiwan University, Taipei, Taiwan; Department of Pharmacy, National Taiwan University Hospital, Taipei, Taiwan. Electronic address: kuoch@ntu.edu.tw.

PMID: 28993139
DOI: 10.1016/j.ab.2017.10.002

Abstract

Glutamine, glutamate and glutathione are key modulators of excessive oxidative stress in tumor cells. In this study, we developed a rapid and accurate HILIC-MS/MS method to simultaneously determine concentrations of cellular glutamine, glutamate and glutathione. A bared silica HILIC column was employed to analyze these polar metabolites. The LC-MS parameters were optimized to achieve high sensitivity and selectivity. The analysis can be completed within 4 min under optimal conditions. The method was validated in terms of accuracy, precision, and linearity. Intra-day (n = 9) precision was within 2.68-6.24% among QCs. Inter-day precision (n = 3) was below 12.4%. The method accuracy was evaluated by the recovery test, and the accuracy for three analytes were between 91.6 and 110%. The developed method was applied to study antioxidant function of GLS2 in non-small cell lung cancer cells. Changes in concentrations of glutamine, glutamate and glutathione revealed that the overexpression of GLS2 could effectively decrease oxidative stress. In summary, this study developed a rapid HILIC-MS/MS method for quantification of GLS2-related metabolites that could facilitate elucidation of the role of GLS2 in tumor development.

Keywords: Cellular metabolites; Glutaminase 2; HILIC-MS/MS; Oxidative stress; Quantification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line, Tumor
Chromatography, High Pressure Liquid*
Glutamic Acid / analysis*
Glutamic Acid / isolation & purification
Glutaminase / metabolism*
Glutamine / analysis*
Glutamine / isolation & purification
Glutathione / analysis*
Glutathione / isolation & purification
Humans
Hydrophobic and Hydrophilic Interactions
Limit of Detection
Tandem Mass Spectrometry*

Substances

Glutamine
Glutamic Acid
Glutaminase
Glutathione