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

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)39-44
Number of pages6
JournalAnalytical Biochemistry
Volume539
DOIs
Publication statusPublished - Dec 15 2017

Keywords

  • Cellular metabolites
  • Glutaminase 2
  • HILIC-MS/MS
  • Oxidative stress
  • Quantification

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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