A stable-isotope HPLC-MS/MS method to simplify storage of human whole blood samples for glutathione assay
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Background: Glutathione is the principal non-protein tripeptide thiol present in most mammalian cells and plays an important role in the redox status of biological systems. The accurate assessment of reduced glutathione (GSH) status as a reliable index of oxidative stress is of research and clinical significance. GSH undergoes rapid oxidation after sample collection and this presents a challenge. Methods: Validation of an HPLC-MS/MS assay is reported. Storage stability using four variants of a methanolic precipitation with addition of stable isotope internal standard at collection is compared to L-serine borate/EDTA with perchloric acid precipitation (SBPE). Results: Precipitation with methanol and addition of stable isotope on sample collection, combined with storage in solution at -70 degrees C showed superior storage stability to SBPE and other variants of the methanolic precipitation method up to 99 days. Conclusions: The combination of stable isotope with methanolic precipitation at collection, with assay by HPLC-MS/MS provides superior results after storage of whole blood samples for at least 99 days. (C) 2012 Elsevier B.V. All rights reserved.
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JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2012, 898 pp. 136 - 140
ELSEVIER SCIENCE BV