Determination of intracellular glutathione and cysteine using HPLC with a monolithic column after derivatization with monobromobimane |
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| Authors: | Xavier A Conlan Nicole Stupka Geoffrey P McDermott Paul S Francis Neil W Barnett |
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| Institution: | 1. Institute for Technology Research and Innovation, Deakin University, Geelong, Victoria 3217 Australia;2. School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217 Australia |
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| 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. |
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| Keywords: | glutathione cysteine monobromobimane human skeletal muscle cells monolithic column chromatography |
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