Comprehensive Lipidome Analysis by Shotgun Lipidomics on a Hybrid Quadrupole-Orbitrap-Linear Ion Trap Mass Spectrometer
Comprehensive Lipidome Analysis by Shotgun Lipidomics on a Hybrid Quadrupole-Orbitrap-Linear Ion Trap Mass Spectrometer
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DOI：
10.1007/s13361-014-1013-x
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发表时间：
2015-01-01
影响因子：
3.2
通讯作者：
Ejsing, Christer S.
中科院分区：
文献类型：
--
作者：
Almeida, Reinaldo;Pauling, Josch Konstantin;Ejsing, Christer S.
在这里，我们报告了一种新的霰弹枪脂质组学平台的应用，该平台具有配备自动化纳米电喷雾离子源的Orbitrap聚变质谱仪。为了评估该平台在深度脂质组分析方面的性能，我们评估了各种仪器参数，包括其无法超越的高分辨率功率，其动态量化范围以及通过基于四极杆的高能碰撞解离（HCD）和基于离子阱的共振激发碰撞诱导解离（CID）对分子脂质的深度结构表征的有效性。该评估表明，采用45万分辨率设置的FTMS分析可以区分来自不同脂质物种的同位素，并具有至少四个数量级的线性动态量化范围。片段分析的评价表明，HCD和CID的联合使用产生了互补的分子脂质片段离子。为了支持全球脂质组分析，我们设计了一种称为MSALL的方法，采用高分辨率FTMS分析进行脂质定量，FTMS2分析使用HCD和CID， ITMS3分析使用双CID进行甘油磷脂分子的深入结构表征。在小鼠小脑和海马的对比分析中，对MSALL方法的性能进行了基准测试。该分析展示了广泛的脂质组定量，涵盖311种脂质，涵盖20个脂类类别，并在应用新的高置信度过滤策略时鉴定出202种不同的甘油磷脂分子。本文提出的工作验证了Orbitrap融合质谱仪用于深入脂质组分析的性能。
Here we report on the application of a novel shotgun lipidomics platform featuring an Orbitrap Fusion mass spectrometer equipped with an automated nanoelectrospray ion source. To assess the performance of the platform for in-depth lipidome analysis, we evaluated various instrument parameters, including its high resolution power unsurpassed by any other contemporary Orbitrap instrumentation, its dynamic quantification range and its efficacy for in-depth structural characterization of molecular lipid species by quadrupole-based higher-energy collisional dissociation (HCD), and ion trap-based resonant-excitation collision-induced dissociation (CID). This evaluation demonstrated that FTMS analysis with a resolution setting of 450,000 allows distinguishing isotopes from different lipid species and features a linear dynamic quantification range of at least four orders of magnitude. Evaluation of fragmentation analysis demonstrated that combined use of HCD and CID yields complementary fragment ions of molecular lipid species. To support global lipidome analysis, we designed a method, termed MSALL, featuring high resolution FTMS analysis for lipid quantification, and FTMS2 analysis using both HCD and CID and ITMS3 analysis utilizing dual CID for in-depth structural characterization of molecular glycerophospholipid species. The performance of the MSALL method was benchmarked in a comparative analysis of mouse cerebellum and hippocampus. This analysis demonstrated extensive lipidome quantification covering 311 lipid species encompassing 20 lipid classes, and identification of 202 distinct molecular glycerophospholipid species when applying a novel high confidence filtering strategy. The work presented here validates the performance of the Orbitrap Fusion mass spectrometer for in-depth lipidome analysis.