A stable-isotope HPLC-MS/MS method to simplify storage of human whole blood samples for glutathione assay

作者：Norris R L G^*; Paul M; George R; Moore A; Pinkerton R; Haywood A; Charles B

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2012, 898: 136-140.

DOI：10.1016/j.jchromb.2012.04.003

• Glutathione
• GSH
• Assay
• Stability
• HPLC-MS/MS

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. %26lt;br%26gt;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). %26lt;br%26gt;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. %26lt;br%26gt;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.

• 出版日期2012-6-1