Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7615790 | Journal of Chromatography B | 2017 | 8 Pages |
Abstract
A simple “one-pot” derivatization and liquid-liquid extraction (LLE) procedure was developed for GC-MS analysis of reduced glutathione (GSH) analysis in erythrocytes. The metabolite was extracted by 5% (w/v) TCA, the supernatant treated with ECF and ethanol-pyridine media, the derivative separated and detected by gas chromatography-mass spectrometry using a short non-polar capillary GC column at a high column-head pressure. Total analysis time was 11 min. The process was optimized by a Design of Experiment. The method was validated showing a good linearity over the 25.4-813.4 μM concentration range, providing satisfactory results in terms of intra-day and inter-day precision as well as an optimal accuracy. The new method was evaluated in a pilot study involving patients with severe protein malnutrition. Comparison of this group with a group of healthy subjects revealed significantly lower GSH concentrations in erythrocytes in the former, thus proving that the described GC-MS method could be employed for fast and simple GSH analysis in clinical studies.
Related Topics
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Analytical Chemistry
Authors
Alessandra Manca, Eugenio Alladio, Paola Massarenti, M. Paola Puccinelli, Antonella De Francesco, Erika Del Grosso, Giulio Mengozzi, Marco Pazzi, Marco Vincenti,