A liquid chromatography/tandem mass spectrometry method for measuring the in vivo incorporation of plasma free fatty acids into intramyocellular ceramides in humans.
A liquid chromatography/tandem mass spectrometry method for measuring the in vivo incorporation of plasma free fatty acids into intramyocellular ceramides in humans.
复制标题
一种液相色谱/串联质谱法，用于测量人体中血浆无脂肪酸的体内掺入人体中细胞内神经酰胺中。
DOI：
10.1002/rcm.6216
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发表时间：
2012-05-15
影响因子：
2
通讯作者：
Jensen, Michael D.
中科院分区：
文献类型：
--
作者：
Blachnio-Zabielska, Agnieszka U.;Persson, Xuan-Mai T.;Koutsari, Christina;Zabielski, Piotr;Jensen, Michael D.
鞘脂是细胞膜的重要组成部分，作为细胞信号分子;神经酰胺在鞘脂代谢中起核心作用。从头神经酰胺生物合成取决于脂肪酸的可用性，但肌肉是否使用循环游离脂肪酸或预先存在的细胞内储存是未知的。我们的目标是开发一种方法来检测静脉注射[U-13 C]棕榈酸酯掺入肌内神经酰胺。我们使用液相色谱/串联质谱法（LC/MS/MS）来测量不同鞘脂种类的浓度和16：0-神经酰胺的13 C同位素富集。使用超高效液相色谱进行色谱分离。采用三重四极杆质谱仪，正离子电喷雾电离源，选择反应监测（SRM）。除富集的神经酰胺外，鞘脂离子以[M+2+H]+形式监测。[13 C16]16：0-神经酰胺监测为[M+16+H]+。通过监测[13 C16]16：0-神经酰胺（554/536和554/264）的2种不同跃迁，我们可以间接测量不是鞘氨醇碱一部分的棕榈酸酯的富集。可使用从接受低剂量[U-13 C]棕榈酸酯输注的志愿者中获得的20 mg肌肉测量浓度和富集。LC/MS/MS可用于体内检测血浆棕榈酸酯掺入人体肌肉神经酰胺的情况。
Sphingolipids are important components of cell membranes that serve as cell signaling molecules; ceramide plays a central role in sphingolipid metabolism. De novo ceramide biosynthesis depends on fatty acid availability, but whether muscle uses circulating free fatty acids or pre-existing intracellular stores is unknown. Our goal was to develop a method to detect the incorporation of intravenously infused [U-13C]palmitate into intramyocellular ceramides. We used a liquid chromatography/tandem mass spectrometry (LC/MS/MS) to measure the concentrations of different sphingolipids species and 13C isotopic enrichment of 16:0-ceramide. Chromatographic separation was performed using ultra performance liquid chromatography. The analysis was performed on triple quadrupole mass spectrometer using positive ion electrospray ionization source with selected reaction monitoring (SRM). The sphingolipids ions, except enriched ceramide, were monitored as [M+2+H]+. The [13C16]16:0-ceramide was monitored as [M+16+H]+. By monitoring 2 different transitions of the [13C16]16:0-Ceramide (554/536 and 554/264) we could indirectly measure enrichment of the palmitate that is not a part of the sphingoid base. Concentration and enrichment could be measured using 20 mg of muscle obtained from volunteers receiving a low dose [U-13C]palmitate infusion. LC/MS/MS can be used to detect the incorporation of plasma palmitate into muscle ceramides in humans, in vivo.
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影响因子：
8.2
作者：
Krssak, M;Petersen, KF;Roden, M
通讯作者：
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