The determination of ascorbic acid and uric acid in human seminal plasma using an HPLC with UV detection
The determination of ascorbic acid and uric acid in human seminal plasma using an HPLC with UV detection
复制标题
DOI：
10.1016/j.jchromb.2011.08.007
复制
发表时间：
2011-09-15
影响因子：
3
通讯作者：
Hampl, Radek
中科院分区：
文献类型：
--
作者：
Kand'ar, Roman;Drabkova, Petra;Hampl, Radek
氧化应激被认为是男性不育的潜在原因之一。抗坏血酸和尿酸在保护精子免受自由基损伤中起重要作用。采用高效液相色谱-紫外检测法同时测定人精浆中抗坏血酸和尿酸的含量，并探讨其作为抗氧化剂保护男性生殖细胞免受氧化损伤的临床意义。精液样本从连续的男性伴侣的夫妇提出生育力评估。液化后，离心样品，用二硫苏糖醇溶液稀释上清液，过滤后注入分析柱。对于分离，使用反相柱MAG 1，250 mm × 4.6 mm，Labiospher PSI 100 C18，5 μ m。以乙醇和25 mmol/L磷酸二氢钠（2.5：97.5，v/v）（pH 4.70）为移动的流动相。该方法对抗坏血酸和尿酸的分析性能令人满意：批内和批间变异系数低于10%。加标精浆的定量回收率在92.1和102.1%之间。我们发现吸烟者和非吸烟者的抗坏血酸和尿酸浓度无显著差异（351.0 +/- 237.9 μ mol/L和323.7 +/- 99.5 μ mol/L vs. 444.8 +/- 245.5 μ mol/L和316.6 +/- 108.9 μ mol/L，p > 0.05）。该方法简便、重复性好，可同时测定人精浆中抗坏血酸和尿酸。(C)2011爱思唯尔有限公司版权所有。
Oxidative stress has been proposed as one of the potential causes for infertility in men. Ascorbic acid and uric acid play important role in protection of spermatozoa against free radicals. A method for the simultaneous determination of ascorbic acid and uric acid in human seminal plasma using HPLC with UV detection and investigation their clinical significance as antioxidants protecting male germ cells against oxidative damage are described. Semen samples were obtained from consecutive male partners of couples presenting fora fertility evaluation. After liquefaction, the samples were centrifuged and the supernatants were diluted with dithiothreitol solution and after a filtration injected onto an analytical column. For the separation, a reverse-phase column MAG 1, 250 mm x 4.6 mm, Labiospher PSI 100 C18, 5 mu m, was used. The mixture of ethanol and 25 mmol/L sodium dihydrogenphosphate (2.5:97.5, v/v), pH 4.70 was used as a mobile phase. Analytical performance of this method is satisfactory for both ascorbic acid and uric acid: the intra-assay and inter-assay coefficients of variation were below 10%. Quantitative recoveries from spiked seminal plasma were between 92.1 and 102.1%. We have found no significant differences in both ascorbic acid and uric acid concentration between the smokers and non-smokers (351.0 +/- 237.9 mu mol/L and 323.7 +/- 99.5 mu mol/L vs. 444.8 +/- 245.5 mu mol/L and 316.6 +/- 108.9 mu mol/L, p > 0.05). This assay is a simple and reproducible HPLC method for the simultaneous measurement of ascorbic acid and uric acid in human seminal plasma. (C) 2011 Elsevier B.V. All rights reserved.