液相串联质谱法定量检测高密度脂蛋白中脂质类化合物

Quantitative determination of lipid compounds in high-density lipoprotein by liquid chromatography tandem mass spectrometry

目的本文旨在建立一种快速提取高密度脂蛋白（HDL）中脂质并结合液相串联质谱定性及快速定量检测的方法。方法采用酸化甲醇沉淀结合超声萃取的方法抽提脂质,高速离心去除蛋白颗粒,取上清液,直接采用液相串联质谱在多反应监测模式下进行定量分析。结果该分离提取脂质的方法简单迅速,采用多反应监测模式可同时定量监测30多种具有标准品的脂质成份,包括鞘氨醇类、鞘磷脂、神经酰胺和氧化型的卵磷脂。结论酸化甲醇超声抽提离心法可简便迅速地获得HDL中的脂质成份,结合液相串联质谱可对HDL中的多种脂质进行定量检测,该方法具有高效、重现性好等优点,适用于大量液体生物样本的分析。

Aim To establish a simplified method for extraction of lipids from high density lipoprotein（ HDL）,qualify and quantify lipids by liquid chromatography tandem mass spectrometry（ LC-MS/MS）. Methods The acidic methanol precipitation combined with ultrasonic extraction method was used to extract lipids,and the supernatant after centrifugation was analyzed directly by LC-MS/MS with multiple reaction monitoring mode. Results The acidic extraction method was effective and time saving,and the established LC-MS/MS method using lipids standards could simultaneously quantify more than 30 kinds of lipids,including sphingolipid,sphigomyelin,ceramide and oxidized phosphatidylcholine.Conclusions Acidic methanol precipitation combined with ultrasonic extraction can effectively isolate lipids from HDL,and the extract after centrifugation can be determined by LC-MS/MS directly. This method is effective with good reproducibility,and is suitable for quick quantitative analysis of a great deal of biological liquid samples.