Sun, Xueni and Heinrich, Paul and Berger, Raffaela S. and Oefner, Peter J. and Dettmer, Katja (2019) Quantification and C-13-Tracer analysis of total reduced glutathione by HPLC-QTOFMS/MS. ANALYTICA CHIMICA ACTA, 1080. pp. 127-137. ISSN 0003-2670, 1873-4324
Full text not available from this repository. (Request a copy)Abstract
Glutathione is an essential intra- and extracellular antioxidant. The level of glutathione in the body is highly related to different disease states and is a useful indicator of disease risk and oxidative stress status. We have developed a sensitive, selective, and comprehensive LC-MS/MS method for the absolute quantification and C-13-tracer analysis of total glutathione using dithiothreitol for the reduction of glutathione disulfide. The limit of detection (LOD) was 0.01 mu M, while the lower limit of quantification (LLOQ) was 0.78 mu M, with the linear (R = 0.9997) range extending up to 100 mu M. The intra-run and inter-run coefficients of variation of 2.49% and 2.04%, respectively, attest to high repeatability. Mean (+/- SD) recoveries of three different concentrations (low, medium, high) of GSH spiked into aliquots of HCT116 cells prior to cell extraction were 108.9% (+/- 2.1), 100.8% (+/- 8.3), and 99.9% (+/- 7.1), respectively. Finally, using a 20 Da wide Q1 window in MRM mode, we were able to detect and relatively quantify all isotopic labeling states of GSH extracted from HCT116 cells fed with either C-13-labeled glucose or glutamine. (C) 2019 Elsevier B.V. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PERFORMANCE LIQUID-CHROMATOGRAPHY; LC-MS/MS METHOD; ISOTOPOMER DISTRIBUTIONS; OXIDIZED GLUTATHIONE; TOTAL HOMOCYSTEINE; HIGH-RESOLUTION; HUMAN PLASMA; QUANTITATION; CYSTEINE; ASSAY; Total glutathione quantification; Reduction; Dithiothreitol; Glutathione isotope labeling pattern; HPLC; Tandem mass spectrometry |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Institut für Funktionelle Genomik > Lehrstuhl für Funktionelle Genomik (Prof. Oefner) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 24 Mar 2020 12:27 |
| Last Modified: | 24 Mar 2020 12:27 |
| URI: | https://pred.uni-regensburg.de/id/eprint/25849 |
Actions (login required)
 |
View Item |
