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

Summary 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).