谷胱甘肽
化学
色谱法
高氯酸
谷胱甘肽二硫化物
三氯乙酸
高效液相色谱法
定量分析(化学)
生物化学
酶
有机化学
作者
Miguel Asensi,Juan Sastre,Federico V. Pallardó,J.G. Delaasuncion,José M. Estrela,José Viña
标识
DOI:10.1006/abio.1994.1126
摘要
A high-performance liquid chromatography method to determine oxidized glutathione (GSSG) in biological samples with ultraviolet-visible detection using N-ethylmaleimide to prevent reduced glutathione (GSH) oxidation is described. Previous methods based on high-performance liquid chromatography to quantitative GSH and GSSG are unsuitable for determining GSSG in biological samples. This is due to GSH oxidation during sample processing. N-Ethylmaleimide, but not iodacetic acid, prevents this oxidation. Blood GSH oxidation measured by the widely used method of Reed et al. (Anal. Biochem. 106, 55-62, 1980) can be as high as 24 +/- 6% (n = 6). When blood samples were assayed by our procedure, GSH oxidation was only 0.13 +/- 0.28% (n = 5). GSH can be determined enzymatically, i.e., with glutathione-S-transferase, but perchloric acid should not be used to deproteinize samples. Trichloroacetic acid (15% final concentration) may be used. This method allows an accurate calculation of the GSH/GSSG ratio, which is important for determining oxidative stress in tissues in various pathophysiological situations.
科研通智能强力驱动
Strongly Powered by AbleSci AI