Department of Clinical Laboratory Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan.
Nihon Waters K.K., Tokyo, Japan.
Sci Rep. 2024 Mar 8;14(1):5699. doi: 10.1038/s41598-024-56321-w.
Mass spectrometry-based lipidomics approaches offer valuable tools for the detection and quantification of various lipid species, including sphingolipids. The present study aimed to develop a new method to simultaneously detect various sphingolipid species that applies to diverse biological samples. We developed and validated a measurement system by employing a single-column liquid chromatography-mass spectrometry system utilizing a normal-phase separation mode with positive ionization. The measurement system provided precision with a coefficient of variant below 20% for sphingolipids in all types of samples, and we observed good linearity in diluted serum samples. This system can measure the following sphingolipids: sphingosine 1-phosphate (S1P), sphingosine (Sph), dihydroS1P (dhS1P), dihydroSph (dhSph), ceramide 1-phosphate (Cer1P), hexosylceramide (HexCer), lactosylceramide (LacCer), dh-ceramide, deoxy-ceramide, deoxy-dh-ceramide, and sphingomyelin (SM). By measuring these sphingolipids in cell lysates where S1P lyase expression level was modulated, we could observe significant and dynamic modulations of sphingolipids in a comprehensive manner. Our newly established and validated measurement system can simultaneously measure many kinds of sphingolipids in biological samples. It holds great promise as a valuable tool for laboratory testing applications to detect overall modulations of sphingolipids, which have been proposed to be involved in pathogenesis processes in a series of elegant basic research studies.
基于质谱的脂质组学方法为检测和定量各种脂质种类，包括鞘脂，提供了有价值的工具。本研究旨在开发一种新方法，以同时检测适用于各种生物样本的各种鞘脂种类。我们开发并验证了一种测量系统，该系统采用正离子化的正相分离模式的单柱液相色谱-质谱系统。该测量系统为所有类型样本中的鞘脂提供了低于 20%变异系数的精度，并且我们在稀释的血清样本中观察到良好的线性关系。该系统可以测量以下鞘脂：1-磷酸鞘氨醇（S1P）、鞘氨醇（Sph）、二氢 S1P（dhS1P）、二氢鞘氨醇（dhSph）、神经酰胺 1-磷酸（Cer1P）、己糖神经酰胺（HexCer）、乳糖基神经酰胺（LacCer）、dh-神经酰胺、脱氧神经酰胺、脱氧-dh-神经酰胺和神经鞘磷脂（SM）。通过测量 S1P 裂解酶表达水平调节的细胞裂解物中的这些鞘脂，我们可以全面观察鞘脂的显著和动态调节。我们新建立和验证的测量系统可以同时测量生物样本中的多种鞘脂。它有望成为实验室检测应用的有价值工具，用于检测一系列优雅的基础研究中提出的鞘脂的整体调节，这些调节被认为与发病机制过程有关。
