Crick Peter J, Bentley T William, Wang Yuqin, Griffiths William J
College of Medicine, Swansea University, Grove Building, Singleton Park, Swansea, SA2 8PP, UK.
Anal Bioanal Chem. 2015 Jul;407(17):5235-9. doi: 10.1007/s00216-015-8609-2. Epub 2015 Mar 22.
Sterols, and specifically oxysterols, play important roles in the biosynthesis of bile acids and steroid hormones as well as possessing biological activities in their own right. Analysis of oxysterols is complicated due to their low abundance in biological systems and poor ionisation characteristics in mass spectrometry. Over the past decade, we have developed a liquid chromatography-mass spectrometry method termed enzyme-assisted derivatisation for sterol analysis (EADSA). Our derivatisation procedure relies on two solid-phase extraction steps to (i) separate cholesterol from oxysterols and (ii) remove excess derivatisation reagents. Recent inter-batch variation in C18 reversed-phase cartridges has led us to experiment with alternative columns. Here, we present our findings and report an improved sample preparation procedure using polymeric hydrophilic-lipophilic balanced reversed-phase cartridges.
甾醇，特别是氧化甾醇，在胆汁酸和类固醇激素的生物合成中发挥着重要作用，并且自身也具有生物活性。由于氧化甾醇在生物系统中的丰度较低以及在质谱分析中电离特性较差，对其进行分析较为复杂。在过去十年中，我们开发了一种液相色谱 - 质谱方法，称为甾醇分析的酶辅助衍生化法（EADSA）。我们的衍生化程序依赖于两个固相萃取步骤，以（i）将胆固醇与氧化甾醇分离，以及（ii）去除过量的衍生化试剂。最近，C18反相柱的批次间差异促使我们试验替代柱。在此，我们展示我们的研究结果，并报告一种使用聚合物亲水亲脂平衡反相柱的改进样品制备程序。
