A stable-isotope HPLC–MS/MS method to simplify storage of human whole blood samples for glutathione assay
Author(s)
Norris, RLG
Paul, M
George, R
Moore, A
Pinkerton, R
Haywood, A
Charles, B
Griffith University Author(s)
Year published
2012
Metadata
Show full item recordAbstract
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 ...
View more >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 ?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.
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View more >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 ?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.
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Journal Title
Journal of Chromatography B
Volume
898
Subject
Analytical chemistry
Biochemistry and cell biology
Pharmacology and pharmaceutical sciences
Pharmaceutical sciences