Assay of total glutathione and glutathione disulphide in seminal plasma of male partners of couples presenting for a fertility evaluation

被引:9
作者
Kand'ar, R. [1 ]
Hajkova, N. [1 ]
机构
[1] Univ Pardubice, Fac Chem Technol, Dept Biol & Biochem Sci, Pardubice 53210, Czech Republic
关键词
Antioxidants; glutathione; glutathione disulphide; human seminal plasma; infertility; oxidative stress; PERFORMANCE LIQUID-CHROMATOGRAPHY; OXIDIZED GLUTATHIONE; REDUCED GLUTATHIONE; QUANTITATIVE-DETERMINATION; FLUORESCENCE DETECTION; MASS-SPECTROMETRY; OXIDATIVE STRESS; WHOLE-BLOOD; AMINOTHIOLS; THIOLS;
D O I
10.1111/and.12176
中图分类号
R69 [泌尿科学(泌尿生殖系疾病)];
学科分类号
摘要
A method is described here for the determination of total glutathione (TGSH) and glutathione disulphide (GSSG) in the seminal plasma of the male partners of couples requesting a fertility evaluation. A suitable sample preparation procedure prior to high-performance liquid chromatography analysis is discussed. After adequate sample preparation, the samples were derivatised with ortho-phthaldialdehyde to form a stable, highly fluorescent tricyclic derivative. Reversed-phase column chromatography was used for the separation, and the effluent was monitored with a fluorescence detector at an excitation wavelength of 350nm and an emission wavelength of 420nm. The analytical performance of this method was satisfactory. The intra-assay and inter-assay coefficients of variation were below 10%. The recoveries were as follows: 94.1% (CV 2.3%) for TGSH and 93.2% (CV 4.0%) for GSSG. No significant differences were found in either TGSH or GSSG concentration between the smokers and nonsmokers (2.07 +/- 1.28m versus 1.56 +/- 1.20m, P=0.431 and 95 +/- 56nm versus 112 +/- 138nm, P=0.825).
引用
收藏
页码:1079 / 1088
页数:10
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