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Determination of intracellular glutathione and cysteine using HPLC with a monolithic column after derivatization with monobromobimane
Author(s) -
Conlan Xavier A.,
Stupka Nicole,
McDermott Geoffrey P.,
Francis Paul S.,
Barnett Neil W.
Publication year - 2010
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1327
Subject(s) - derivatization , chemistry , glutathione , high performance liquid chromatography , chromatography , cysteine , quantitative analysis (chemistry) , biochemistry , enzyme
Abstract An optimized high‐performance liquid chromatography (HPLC) method is used to show that, as myoblasts differentiate into multinucleated muscle fibers, there is a shift to a more oxidized cell redox state. The HPLC method incorporated derivatization with monobromobimane for the determination of the reduced (GSH) and oxidized (GSSG) forms of glutathione and the reduced (Cys) and oxidized (CysSS) forms of cysteine. The derivatization was optimized to improve the sensitivity of the approach; the limits of detection for glutathione and cysteine were 3 × 10 −8 and 5 × 10 −8 M , respectively. Copyright © 2009 John Wiley & Sons, Ltd.