光谱学与光谱分析 |
|
|
|
|
|
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.
|
Received: 2004-06-28
Accepted: 2004-09-08
|
|
Corresponding Authors:
WANG Shu-ren
|
|
Cite this article: |
WANG Qiu-lin,WANG Hao-yi,DING Yi, et al. A Novel Fluorophotometric Method for Assaying Free Glutathione of Both Reduced and Oxidized in Plasma [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2005, 25(11): 1834-1838.
|
|
|
|
URL: |
https://www.gpxygpfx.com/EN/Y2005/V25/I11/1834 |
[1] Smith C V, Jones D P, Guenthner T M, et al. Tox. Appl. Pharmacol., 1996, 140: 1. [2] McNeil T L, Beck L V. Analytical Biochemistry, 1968, 22: 431. [3] Cereser C, Guichard J, Drai J, et al. Journal of Chromatography B, 2001, 752: 123. [4] Cotgreave I A, Moldéus P. J. Biochem. Biophys. Methods, 1986, 13(4-5): 231. [5] Chong C Y, Huxtable R J. Journal of Chromatography B, 1995, 672(2): 217. [6] Morineau G, Azoulay M, Frappier F. Journal of Chromatography A, 1989, 467(1): 209. [7] Cohn V H, Lyle J. Analytical Biochemistry, 1966, 14: 434.
|
[1] |
QIAO Lu, WANG Song-lei*, GUO Jian-hong, HE Xiao-guang. Combination of Spectral and Textural Informations of Hyperspectral Imaging for Predictions of Soluble Protein and GSH Contents in Mutton[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2022, 42(01): 176-183. |
[2] |
ZHAO Hui-hui1,HOU Na2, WANG Wei2* . Difference Expressed Protein Study on Unstable Angina Blood-Stasis Syndrome by Fluorescence Labelling Method[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2009, 29(06): 1647-1650. |
[3] |
ZHANG Lin-lin1,XING Xiao-ru2,WU Guo-ping3,WEI Fu-sheng3* . Determination of Thirty Trace Elements in Human Plasma by Microwave Digestion-ICP-MS[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2009, 29(04): 1115-1118. |
[4] |
LI Wei1,2,KE Yong1,HE Guang-she3,XU Yong-cheng1,WANG Zhen-yuan1* . Study on the Correlation between PMI and OD Changes in Rat’s Plasma[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2008, 28(12): 2944-2946. |
[5] |
LONG Wen-qing. Determination of Trace Vitamin B2 by Fluorimetry after Aqueous Two-Phase Extraction Based on Acetone and Ethanol[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2007, 27(10): 2098-2101. |
[6] |
ZHANG Xiao-dong1, ZHAO Yuan-li2, GUO Tao3, WANG Lei-ming4, GE Xiang-hong2, LUO Lei2 . Study on Absorption Spectra of Malignant Plasma of Esophagus[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2006, 26(08): 1533-1535. |
[7] |
DENG Jian1, XU Jin-sheng2, FAN Hua1, YUAN Ya-li1, LIU Chuan-xiang1 . Direct Determination of Ammonia in Plasma with Indophenol Method [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2005, 25(03): 436-438. |
|
|
|
|