![Sample our Physical Sciences journals, sign in here to start your FREE access for 14 days]({"type":"image" , "src" : "/sda/110819/TOC-Subject-Sample-2021-Physical-Sciences.jpg"})
Abstract
A method is described here for the determination of total glutathione and glutathione disulfide in human whole blood and plasma with a suitable sample preparation procedure to prevent glutathione oxidation and glutathione disulfide hydrolysis. Whole blood samples were obtained from a group of blood donors. After adequate sample preparation, the samples were derivatized with ortho-phthaldialdehyde to form a stable, highly fluorescent tricyclic derivative. Reverse-phase column chromatography was used for the separation and fluorimetric detection to monitor the effluent. The analytical performance of this method was satisfactory for the determination of total glutathione and glutathione disulfide. The intra-assay and inter-assay coefficients of variation were below 10%. The recoveries were as follows: 96.1% (CV 1.4%) and 104.3% (CV 2.4%) for whole blood, 92.3% (CV 2.4%) and 107.4% (CV 2.6%) for plasma. We found no significant differences in both total glutathione and glutathione disulfide concentration between the women and men. We developed a selective high-performance liquid chromatography method for the determination of total glutathione and glutathione disulfide in human whole blood and plasma. Our sophisticated sample preparation procedure prevents significant glutathione oxidation and glutathione disulfide hydrolysis.
ACKNOWLEDGMENT
This work was supported by Grants SGFChT07/2012 and MSMT 0021627502 from the Ministry of Education.
Notes
a IQR, interquartile range is the difference between the upper quartile and the lower quartile.
a 11-point for the determination of analytical parameters and 7-point for routine analysis.
b The x-intercept (in µmol/L) is the point at which the line crosses the x axis (where the y value equals 0).