鞘脂类中的主要活性分子鞘氨醇1-磷酸(S1P),可通过介导细胞增殖、分化和迁移等发挥广泛的生物学功能;同时,S1P可在相关酶的作用下转变为其它形式.本文旨在建立一种简便的鞘脂类的制备方法,并结合液相串联质谱(LC-MS/MS)快速检测生物样本中纳克水平的鞘脂类化合物.采用甲醇沉淀或经典的脂质提取方法获得鞘脂类化合物,再采用LC-MS/MS在多反应监测模式(MRM)下进行定量分析.实验结果表明,甲醇沉淀法可简便快捷地获得血浆或脂蛋白中鞘氨醇类化合物;S1P、二氢鞘氨醇(DH-S1P)和鞘氨醇(SPH)的定量限分别为110.5、215.6和44.3 pg;人血浆中S1P、DH-S1P和SPH的含量分别为257.8±49.4 nmol/L、93.5±17.3 nmol/L和44.6±7.4nmol/L,鞘脂类化合物在人血浆脂蛋白上的分布存在显著性差异;在ApoE-/-小鼠血浆中S1P、DHS1P和SPH的含量分别为590.1±78.2 nmol/L、197.8±60.6 nmol/L和35.4±16.7 nmol/L;每1×106个人脐静脉内皮细胞(EA.hy926)中,S1P和SPH的含量分别为103.7±21.8 pg和16.3±5.3ng.甲醇沉淀法结合LC-MS/MS可简便快捷地对血浆和脂蛋白中的鞘脂类化合物进行定量检测.该方法特别适用于大量生物样本中鞘脂类的快速定量分析. Sphingosine 1-phosphate (S1P), the major active molecule in sphingolipids, can exert a wide range of biological functions through mediating cell proliferation, differentiation and migration. At the same time, S1P can be transformed into other forms under the action of related enzymes The purpose of this paper is to establish a simple and convenient method for the preparation of sphingolipids, and to rapidly detect nanograms of sphingolipids in biological samples by liquid chromatography-tandem mass spectrometry (LC-MS / MS) The sphingolipids were obtained by mass extraction method and then analyzed quantitatively by multi-reaction monitoring mode (MRM) by LC-MS / MS.The experimental results showed that the sphingosines in plasma or lipoprotein could be obtained easily and rapidly by methanol precipitation method. The limit of quantification of S1P, sphinganine (DH-S1P) and sphingosine (SPH) were 110.5, 215.6 and 44.3 pg, respectively. The contents of S1P, DH-S1P and SPH in human plasma were 257.8 ± 49.4 nmol / L, 93.5 ± 17.3 nmol / L and 44.6 ± 7.4 nmol / L, respectively. The distribution of sphingolipids in human plasma lipoproteins was significantly different. In the plasma of ApoE - / - mice, the distribution of S1P, DHS1P and SPH The contents were 590.1 ± 78.2 nmol / L, 197.8 ± 60.6 nmol / L and 35.4 ± 16.7 nmol / L, respectively; The contents of S1P and SPH in arterial endothelial cells (EA.hy926) were 103.7 ± 21.8 pg and 16.3 ± 5.3 ng, respectively. Methanol precipitation combined with LC-MS / MS was a simple, rapid and sensitive method for the determination of sphingolipids Compounds for quantitative detection.The method is particularly suitable for rapid quantitative analysis of sphingolipids in a large number of biological samples.