Rapid quantification of glutaminase 2 (GLS2)-related metabolites by HILIC-MS/MS
Journal
Analytical Biochemistry
Journal Volume
539
Pages
39-44
Date Issued
2017
Author(s)
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. ? 2017
SDGs
Other Subjects
glutamic acid; glutaminase; glutaminase 2; glutamine; glutathione; unclassified drug; glutamic acid; glutaminase; glutamine; glutathione; accuracy; Article; controlled study; human; human cell; hydrophilic interaction chromatography; liquid chromatography-mass spectrometry; lung non-small cell carcinoma cell line; mass spectrometry; metabolite; oxidative stress; priority journal; protein expression; quantitative analysis; chemical phenomena; high performance liquid chromatography; isolation and purification; limit of detection; metabolism; tandem mass spectrometry; tumor cell line; Cell Line, Tumor; Chromatography, High Pressure Liquid; Glutamic Acid; Glutaminase; Glutamine; Glutathione; Humans; Hydrophobic and Hydrophilic Interactions; Limit of Detection; Tandem Mass Spectrometry
Type
journal article