Quantitative analysis of desmosterol, cholesterol and cholesterol sulfate in semen by high-performance liquid chromatography
Quantitative analysis of desmosterol, cholesterol and cholesterol sulfate in semen by high-performance liquid chromatography
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DOI：
10.1016/s0021-9673(01)01105-0
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发表时间：
2001-11-23
影响因子：
4.1
通讯作者：
Boucher, D
中科院分区：
文献类型：
--
作者：
Sion, B;Grizard, G;Boucher, D
本文介绍了一种简便、快速、准确地分离测定人精子和精浆中胆固醇、链甾醇和硫酸胆固醇的方法。该高效液相色谱法基于Inertsil ODS 2 5 μ m硅胶柱上的反相色谱法，以氯仿-甲醇和氯仿-甲醇-水混合物的二元梯度作为移动的相，流速为0.25 ml/min。分离度良好且重现性高。使用光散射（质量）检测器定量洗脱的甾醇。在这些条件下，可以分别定量低至64、64和68 pmol的胆固醇、链甾醇和胆固醇硫酸盐。胆固醇是精子中的主要固醇，（107 +/-7 nmol/10（8）个精子）和SP（0.83 +/-0.10 μ mol/ml），而链甾醇的浓度在精子中为38 +/-6 nmol/10（8），在SP中为0.18 +/-0.02 μ mol/ml。总胆固醇在精子和SP的结论，这种方法提供了有趣的前景，这些甾醇的定量分析不仅在精液中，而且在其他生物样品。(C)2001 Elsevier Science B. V.保留所有权利。
A simple, rapid and accurate method to separate and quantify cholesterol, desmosterol and cholesterol sulfate in human spermatozoa and seminal plasma (SP) is described. This high-performance liquid chromatographic procedure is based on reversed-phase chromatography on a Inertsil ODS2 5 mum silica column with a binary gradient of mixtures of chloroform-methanol and chloroform-methanol-water as the mobile phase at a flow-rate of 0.25 ml/min. Sterols are separated with good resolution and high reproducibility. The eluted sterols are quantified using a light-scattering (mass) detector. As little as 64, 64 and 68 pmol of cholesterol, desmosterol and cholesterol sulfate, respectively, can be quantified under these conditions. Cholesterol is the predominant sterol both in spermatozoa (107 +/-7 nmol/10(8) spermatozoa) and SP (0.83 +/-0.10 mu mol/ml) whereas the concentrations of desmosterol were 38 +/-6 nmol/10(8) in spermatozoa and 0.18 +/-0.02 mu mol/ml in SP Cholesterol sulfate represents about 6% of total cholesterol in the spermatozoa and SP In conclusion, this method offers interesting perspectives for the quantitative analysis of these sterols not only in semen, but also in other biological samples. (C) 2001 Elsevier Science B.V. All rights reserved.