Glutathione (GSH) is a major endogenous antioxidant that has a central role in cellular defense against toxins and free radicals. Rapid and accurate detection of GSH content in single cells is important to the early diagnosis of disease and biomedical research. In this work, a novel method based on microchip electrophoresis chemiluminescence (MCE-CL) detection was developed for the quantification of glutathione (GSH) in single cells from rat liver. The detection of GSH is based on the strong sensitization of mercapto compound to luminol-H2O2CL system. The injection, localization, and membrane dissolution of single cell were simply and rapidly carried out on the microchip by direct electric field force, which did not require any additional membrane dissolution reagent. Under optimized experimental conditions, single cell assay was achieved within 2min. The peak area of the GSH was taken as quantification of GSH, and a good linear relationship of GSH concentration to peak area in the range of 3.0 × 10-6M to 6.0 × 10-4M was obtained. The detection limit for GSH is 9.6 × 10-7M, calculated by S/N = 3. The measured GSH content in single cells from rat liver (n = 10) ranged from 7.8fmol to 13.fmol with a mean value of 10.8fmol.
Quantification of glutathione in single cells from rat liver by microchip electrophoresis with chemiluminescence detection.