Simultaneous quantification of cholesterol sulfate, androgen sulfates, and progestagen sulfates in human serum by LC-MS/MS
Simultaneous quantification of cholesterol sulfate, androgen sulfates, and progestagen sulfates in human serum by LC-MS/MS
复制标题
DOI：
10.1194/jlr.d061499
复制
发表时间：
2015-09-01
影响因子：
6.5
通讯作者：
Wudy, Stefan A.
中科院分区：
文献类型：
--
作者：
Sanchez-Guijo, Alberto;Oji, Vinzenz;Wudy, Stefan A.
类固醇主要以硫酸化形式存在于人体体液中。从诊断和生理学的角度来看，这些化合物的分析是重要的。在这里，我们提出了一种通过LC-MS/MS定量人血清中11种完整类固醇硫酸酯的新方法。在我们的方法中分析的化合物，其中一些是首次在血液中定量的，包括胆固醇硫酸酯，硫酸双烯醇酮，17-羟基-硫酸双烯醇酮，16-羟基-硫酸脱氢表雄酮，硫酸脱氢表雄酮，硫酸雄烯二醇，硫酸雄酮，硫酸表雄酮，硫酸睾酮、硫酸表睾酮和硫酸二氢睾酮。该测定被设想为在广泛的浓度范围内定量硫酸化类固醇，仅需要300 L血清。该方法已得到验证，并在三个质量控制下研究了其性能，根据其生理浓度为每种化合物选择。该测定法显示出所有化合物的良好线性（R-2 > 0.99）和回收率，定量限在1和80 ng/ml之间。平均日内和日间精密度（变异系数）和准确度（相对误差）均低于10%。该方法已成功应用于研究类固醇硫酸酯酶缺乏症等疾病中的硫酸化类固醇组，证明了其诊断价值。据我们所知，这是最全面的方法可用于定量人血液中的硫酸化类固醇。
Steroids are primarily present in human fluids in their sulfated forms. Profiling of these compounds is important from both diagnostic and physiological points of view. Here, we present a novel method for the quantification of 11 intact steroid sulfates in human serum by LC-MS/MS. The compounds analyzed in our method, some of which are quantified for the first time in blood, include cholesterol sulfate, pregnenolone sulfate, 17-hydroxy-pregnenolone sulfate, 16--hydroxy-dehydroepiandrosterone sulfate, dehydroepiandrosterone sulfate, androstenediol sulfate, androsterone sulfate, epiandrosterone sulfate, testosterone sulfate, epitestosterone sulfate, and dihydrotestosterone sulfate. The assay was conceived to quantify sulfated steroids in a broad range of concentrations, requiring only 300 l of serum. The method has been validated and its performance was studied at three quality controls, selected for each compound according to its physiological concentration. The assay showed good linearity (R-2 > 0.99) and recovery for all the compounds, with limits of quantification ranging between 1 and 80 ng/ml. Averaged intra-day and between-day precisions (coefficient of variation) and accuracies (relative errors) were below 10%. The method has been successfully applied to study the sulfated steroidome in diseases such as steroid sulfatase deficiency, proving its diagnostic value. This is, to our best knowledge, the most comprehensive method available for the quantification of sulfated steroids in human blood.