使用两种稳定同位素标记的鞘磷脂物种通过液相色谱-质谱联用法定量和定性分析鞘磷脂的方法
Quantitative and Qualitative Method for Sphingomyelin by LC-MS Using Two Stable Isotopically Labeled Sphingomyelin Species.
作者信息
Hama Kotaro, Fujiwara Yuko, Yokoyama Kazuaki
机构信息
Faculty of Pharmaceutical Sciences, Teikyo University;
Faculty of Pharmaceutical Sciences, Teikyo University.
出版信息
J Vis Exp. 2018 May 7(135):57293. doi: 10.3791/57293.
Abstract
We present a method of analyzing sphingomyelin (SM) qualitatively and quantitatively by liquid chromatography-electrospray Ionization-tandem mass spectrometry (LC-ESI-MS/MS). SM is a common sphingolipid composed of a phosphorylcholine and a ceramide as the hydrophilic and hydrophobic component, respectively. A number of SM species are present in mammalian cells due to a variety in the sphingoid long chain base (LCB) and an N-acyl moiety in the ceramide. In this report, we show a method of estimating the number of carbon and double bonds in a LCB and an N-acyl moiety based on their corresponding product ions in MS/MS/MS (MS) experiments. In addition, we present a quantitative analysis method for SM using two stable isotopically labeled SM species, which facilitates determining the range used in SM quantitation. The present method will be useful in characterizing a variety of SM species in biological samples and industrial products such as cosmetics.
摘要
我们提出了一种通过液相色谱-电喷雾电离串联质谱法（LC-ESI-MS/MS）对鞘磷脂（SM）进行定性和定量分析的方法。SM是一种常见的鞘脂，分别由磷酸胆碱和神经酰胺作为亲水和疏水成分组成。由于鞘氨醇长链碱基（LCB）和神经酰胺中的N-酰基部分存在多种变化，哺乳动物细胞中存在多种SM种类。在本报告中，我们展示了一种基于MS/MS/MS（MS）实验中相应的产物离子来估计LCB和N-酰基部分中碳和双键数量的方法。此外，我们提出了一种使用两种稳定同位素标记的SM种类对SM进行定量分析的方法，这有助于确定SM定量中使用的范围。本方法将有助于表征生物样品和化妆品等工业产品中的各种SM种类。
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