Journal article

Determination of intracellular glutathione and cysteine using HPLC with a monolithic column after derivatization with monobromobimane

Xavier A Conlan, Nicole Stupka, Geoffrey P McDermott, Paul S Francis, Neil W Barnett

BIOMEDICAL CHROMATOGRAPHY | JOHN WILEY & SONS LTD | Published : 2010

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.

University of Melbourne Researchers