A Novel Fluorophotometric Method for Assaying Free Glutathione of Both Reduced and Oxidized in Plasma
WANG Qiu-lin1，2, WANG Hao-yi1, DING Yi1, QIAO Xiao-rong3, PENG Ke-jun1, LIN Xia4, WANG Shu-ren1*
1. Department of Pathophysiology, West-China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China 2. The First Department of Internal Medicine, the First Hospital Affiliated to Chengdu Medical College, Chengdu 610500, China 3. Analysis and Testing Center, Sichuan University, Chengdu 610041，China 4. Department of Forensic Pathology, West-China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China
Abstract：To investigate the measurement of free glutathione of both reduced or oxidized (i.e. GSH and GSSG) in plasma, and to evaluate the redox state of GGSH/GSSG in human plasma, both GSH and GSSG in plasma were measured by fluorophotometry based on the facts that one molar GSSG can be reduced to two molar GSH by dithiothreitol(DTT) under the condition of the pH being about 6.0, and GSH can provide both primary amine and thiol groups to react with the two carbonyl groups of O-phthaldehyde(OPA) to form a fluorescent ternary isoindole complex at pH 8.0. This method can at least measure 16 picomole GSH and 8 picomole GSSG respectively in the tube. The variation coefficient (CV) for intra-ssay and intera-ssay is about 4..6% and 3.9% for GSH and 3.5% and 4.1% for GSSG respectively. The recovery of GSH and GSSG added to the plasma is (99.77%±5.70)% and (99.28%±4.73)% respectively. The concentration of GSH and GSSG in the plasma of young healthy volunteer is (16.5±2.4)nmol·mL-1 and (1.7±0.35)nmol·mL-1 respectively, without significant difference between male and female. This measurement method is simple with great sensitivity and selectivity for rapid measuring GSH and GSSG in human plasma simultaneously.
王秋林1, 2，王浩毅1，丁 怡1，乔小蓉3，彭克军1，林 霞4，王树人1* . 血浆中游离还原型及氧化型谷胱甘肽的荧光测定法[J]. 光谱学与光谱分析, 2005, 25(11): 1834-1838.
WANG Qiu-lin1，2, WANG Hao-yi1, DING Yi1, QIAO Xiao-rong3, PENG Ke-jun1, LIN Xia4, WANG Shu-ren1* . A Novel Fluorophotometric Method for Assaying Free Glutathione of Both Reduced and Oxidized in Plasma . SPECTROSCOPY AND SPECTRAL ANALYSIS, 2005, 25(11): 1834-1838.
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