A comprehensive method for extraction and quantitative analysis of sterols and secosteroids from human plasma
A comprehensive method for extraction and quantitative analysis of sterols and secosteroids from human plasma
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DOI：
10.1194/jlr.d022285
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发表时间：
2012-07-01
影响因子：
6.5
通讯作者：
Russell, David W.
中科院分区：
文献类型：
--
作者：
McDonald, Jeffrey G.;Smith, Daniel D.;Russell, David W.
我们描述了一种方法的开发，用于从人血浆中提取和分析62甾醇，氧化甾醇，开环甾类化合物，使用HPLC-MS和GC-MS的组合。氘代标准添加到200 μ l的人血浆。散装脂质提取与甲醇：二氯甲烷，样品水解使用一种新的程序，甾醇和开环甾类化合物分离使用固相萃取（SPE）。化合物在C-18核-壳HPLC柱上和通过GC分离。使用三重四极杆质谱仪测量甾醇和氧化甾醇，并且使用GC-MS测量脂甾醇。通过HPLC-MS测量的每种化合物的检测是布尔OR lng/ml血浆。提取效率在85%和110%之间;日间变异性显示相对标准误差为
We describe the development of a method for the extraction and analysis of 62 sterols, oxysterols, and secosteroids from human plasma using a combination of HPLC-MS and GC-MS. Deuterated standards are added to 200 mu l of human plasma. Bulk lipids are extracted with methanol: dichloromethane, the sample is hydrolyzed using a novel procedure, and sterols and secosteroids are isolated using solid-phase extraction (SPE). Compounds are resolved on C-18 core-shell HPLC columns and by GC. Sterols and oxysterols are measured using triple quadrupole mass spectrometers, and lathosterol is measured using GC-MS. Detection for each compound measured by HPLC-MS was boolean OR 1 ng/ml of plasma. Extraction efficiency was between 85 and 110%; day-to-day variability showed a relative standard error of