title: Assay of total glutathione and glutathione disulphide in seminal plasma of male partners of couples presenting for a fertility evaluation.
abstract: 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 350 nm and an emission wavelength of 420 nm. 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 ± 1.28 μm versus 1.56 ± 1.20 μm, P = 0.431 and 95 ± 56 nm versus 112 ± 138 nm, P = 0.825).
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Pardubice, Czech Republic.
authors.forename: R
authors.initials: R
authors.lastname: Kanďár
authors.forename: N
authors.initials: N
authors.lastname: Hájková
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid; D005978:Glutathione / Q000032:analysis*; D019803:Glutathione Disulfide / Q000032:analysis*; D006801:Humans; D007248:Infertility, Male / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D012661:Semen / Q000737:chemistry*; D055815:Young Adult
chemical_list: D005978:Glutathione; D019803:Glutathione Disulfide
keywords: Antioxidants; glutathione; glutathione disulphide; human seminal plasma; infertility; oxidative stress