在动物血中使用液态染色学-质谱法对多个类型的类固醇进行全面的向分析
Taťána Gazárková1, Hana Kočová Vlčková1, Kateřina Plachká1
1Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Heyrovského 1203, 500 03, Hradec Králové, Czech Republic.
Analytica chimica acta
|November 23, 2024
概括
这项研究开发了一种新的UHPLC-MS/MS方法,用于同时进行类固醇分析,克服了常见的挑战. 经过验证的方法可以在最小的样本体积中准确量化38种类固醇,用于研究和临床使用.
科学领域:
- 分析化学 分析化学
- 生物化学 生物化学
- 内分泌学 在内分泌学.
背景情况:
- 在生物流体中同时量化多个类固醇在分析上具有挑战性,原因是异构体交叉声和矩阵干扰等问题.
- 现有的方法难以同时分析广泛的类固醇,限制了全面的分析.
研究的目的:
- 开发一种无衍生,高吞吐量,具有成本效益的UHPLC-MS/MS方法,用于同时分析38种类固醇.
- 解决分析障碍,包括在类固醇量化中的同位素分离和矩阵效应.
- 根据ICH/EMA M10指南使用新型代用矩阵方法验证该方法.
主要方法:
- 开发了一种无衍生,无性UHPLC-MS/MS方法用于类固醇分析.
- 优化蛋白质沉 (PP) 和支持液体提取 (SLE) 方法,使用小样本体积 (40μL).
- 实施了一种新的代孕矩阵背景减法技术,使用上腺切除的老鼠血.
主要成果:
- 在20分钟的运行时间内成功分辨出16种立体异构体和15种定位异构体,分析了38种类固醇.
- 达到PP和SLE的低量化极限,尽管一些类固醇仅在质量上进行了评估.
- 用代孕矩阵方法验证了UHPLC-MS/MS工作流,在500多只小鼠血样本中证明了其有效性.
结论:
- 精确的类固醇量化通常被低估,损害了研究和临床应用.
- 综合性,经过验证的UHPLC-MS/MS方法减轻了类固醇交叉声和使用最小的样本体积.
- 新的代孕矩阵方法显著推进了用于各种生物应用的类固醇分析.
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