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

Norris, R. L. G., Paul, M., George, R., Moore, A., Pinkerton, R., Haywood, A. and Charles, B. (2012) A stable-isotope HPLC-MS/MS method to simplify storage of human whole blood samples for glutathione assay. Journal of Chromatography B, 898 136-140. doi:10.1016/j.jchromb.2012.04.003

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Author Norris, R. L. G.
Paul, M.
George, R.
Moore, A.
Pinkerton, R.
Haywood, A.
Charles, B.
Title A stable-isotope HPLC-MS/MS method to simplify storage of human whole blood samples for glutathione assay
Journal name Journal of Chromatography B   Check publisher's open access policy
ISSN 1570-0232
1873-376X
Publication date 2012-06-01
Sub-type Article (original research)
DOI 10.1016/j.jchromb.2012.04.003
Volume 898
Start page 136
End page 140
Total pages 5
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2013
Language eng
Formatted abstract 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.

Highlights: ► A n HPLC–MS/MS assay for determination of reduced glutathione is presented. ► Deuterated internal standard with precipitation at sample collection allows storage for 99 days. ► Improved storage compared to EDTA/perchloric acid/serine borate pre-treatment.
Keyword Glutathione
GSH
Assay
Stability
HPLC–MS/MS
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Pharmacy Publications
 
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Created: Thu, 24 May 2012, 14:45:37 EST by Assoc Prof Bruce Charles on behalf of School of Pharmacy