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Elution, Partial Separation, and Identification of Lipids Directly from Tissue Slices on Planar Chromatography Media by Desorption Electrospray Ionization Mass Spectrometry

机译:脱附电喷雾电离质谱法直接从平面层析介质上的组织切片中洗脱,部分分离和鉴定脂质

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摘要

A method for the examination of intact tissue sections for gangliosides and other lipids using desorption electro-spray ionization (DESI) mass spectrometry (MS) is presented. In the present work, thin tissue slices (16 (mu)m) taken from the rat brain are thaw mounted onto planar chromatographic media and the lipids are eluted, partially separated, and analyzed directly from the plate by DESI-MS in the negative ion mode. With the lanes scanned parallel to the direction of the chromatographic separation in the full scan mode, the selected ion current associated with ions of separated lipid molecules is plotted in order of increasing Rfvalues. Distinctly different classes of lipids are detected using this method, including several ganglioside species (i.e., GQ1, GT1, GD1, and GM1) and sulfoglycosphingolipids. For the examination of gangliosides in the full scan negative ion mode from high-performance thin-layer chromatography (HPTLC) plates, the limit of detection (LOD) was determined to be approximately 3 pmol. Tandem mass spectrometry (MS/MS) using the linear ion trap was used to confirm the presence of selected gangliosides and other lipids directly from the HPTLC plate. DESI-MS/MS revealed the presence of both the GD1a and GD1b isomers. The simplicity of this approach where planar separations are relied upon for sample preparation and presentation to the MS should allow for the examination of a variety of complex samples including the rapid examination of foodstuffs, bacteria, whole blood, and needle biopsies for cancer diagnostics.
机译:提出了一种使用解吸电喷雾电离(DESI)质谱(MS)检查完整组织切片中神经节苷脂和其他脂质的方法。在本工作中,将从大鼠大脑中取出的薄组织切片(16μm)解冻安装在平面色谱介质上,并将脂质洗脱,部分分离,并通过DESI-MS在负离子中直接从板上进行分析。模式。在全扫描模式下,平行于色谱分离方向扫描的泳道,将按Rf值递增的顺序绘制与分离的脂质分子的离子相关的选定离子电流。使用此方法可检测到截然不同的类脂,包括几种神经节苷脂种类(即GQ1,GT1,GD1和GM1)和磺基糖鞘脂。为了从高性能薄层色谱(HPTLC)板以全扫描负离子模式检查神经节苷脂,检测限(LOD)确定为大约3 pmol。使用线性离子阱的串联质谱(MS / MS)用于直接从HPTLC板中确认所选神经节苷脂和其他脂质的存在。 DESI-MS / MS揭示了GD1a和GD1b异构体的存在。这种方法的简单性(依靠平面分离进行样品制备并提交给MS)应允许检查各种复杂的样品,包括快速检查食品,细菌,全血和活检针以进行癌症诊断。

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