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

Fingerprint

Glutaminase
Metabolites
Glutamine
Glutathione
Glutamic Acid
Oxidative stress
Cells
Tumors
Oxidative Stress
Silicon Dioxide
Modulators
Non-Small Cell Lung Carcinoma
Antioxidants
Neoplasms
Recovery

Keywords

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

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Rapid quantification of glutaminase 2 (GLS2)-related metabolites by HILIC-MS/MS. / Chen, Guan Yuan; Chao, Hsi Chun; Liao, Hsiao Wei; Tsai, I. Lin; Kuo, Ching Hua.

In: Analytical Biochemistry, Vol. 539, 15.12.2017, p. 39-44.

Research output: Contribution to journalArticle

Chen, Guan Yuan ; Chao, Hsi Chun ; Liao, Hsiao Wei ; Tsai, I. Lin ; Kuo, Ching Hua. / Rapid quantification of glutaminase 2 (GLS2)-related metabolites by HILIC-MS/MS. In: Analytical Biochemistry. 2017 ; Vol. 539. pp. 39-44.
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