The determination of ascorbic acid and uric acid in human seminal plasma using an HPLC with UV detection

摘要: Oxidative stress has been proposed as one of the potential causes for infertility in men. Ascorbic acid and uric play important role protection spermatozoa against free radicals. A method simultaneous determination ascorbic human seminal plasma using HPLC with UV detection investigation their clinical significance antioxidants protecting male germ cells oxidative damage are described. Semen samples were obtained from consecutive partners couples presenting a fertility evaluation. After liquefaction, centrifuged supernatants diluted dithiothreitol solution after filtration injected onto an analytical column. For separation, reverse-phase column MAG 1, 250 mm × 4.6 mm, Labiospher PSI 100 C18, 5 μm, was used. The mixture ethanol 25 mmol/L sodium dihydrogenphosphate (2.5:97.5, v/v), pH 4.70 used mobile phase. Analytical performance this is satisfactory both acid: intra-assay inter-assay coefficients variation below 10%. Quantitative recoveries spiked between 92.1 102.1%. We have found no significant differences concentration smokers non-smokers (351.0 ± 237.9 μmol/L 323.7 99.5 vs. 444.8 245.5 316.6 108.9 μmol/L, p>0.05). This assay simple reproducible measurement plasma.