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Updated: Jun 20, 2026

Rapid Quantification of Oxidized and Reduced Forms of Glutathione Using Ortho -phthalaldehyde in Cultured Mammalian Cells In Vitro
Published on: June 28, 2024
Determination of intracellular glutathione and cysteine using HPLC with a monolithic column after derivatization with
Xavier A Conlan1, Nicole Stupka, Geoffrey P McDermott
1Institute for Technology Research and Innovation, Deakin University, Geelong, Victoria, Australia. xavier@deakin.edu.au <xavier@deakin.edu.au>
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 x 10(-8) and 5 x 10(-8) M, respectively.

