Prioritize biologically relevant ions for data-independent acquisition (BRI-DIA) in LC-MS/MS-based lipidomics analysis
Prioritize biologically relevant ions for data-independent acquisition (BRI-DIA) in LC-MS/MS-based lipidomics analysis
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DOI：
10.1007/s11306-022-01913-8
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发表时间：
2022-07-16
期刊：
影响因子：
3.6
通讯作者：
Liu, Xiaojing
中科院分区：
文献类型：
--
作者：
Duan, Likun;Scheidemantle, Grace;Liu, Xiaojing
数据依赖性采集（DDA）是基于Orbitrap仪器的脂质组学分析中最常用的MS/MS扫描方法。然而，MS仪器相关软件决定用于片段化的前N个前体，导致前体选择的随机性和妥协的一致性和再现性。我们介绍了一种使用生物相关脂质构建数据独立采集（DIA）包含列表的新型工作流程，称为BRI-DIA工作流程。目的确保基于LC-MS/MS的脂质组学分析中生物相关脂质的一致覆盖。方法基于MS-DIAL 4中的LIPID MAPS和脂质组图谱构建生物相关离子列表。结果DDA导致更多的MS/MS事件，但通过三种方法（DDA、BRI-DIA和混合MS/MS扫描模式）鉴定的独特脂质的总数是相当的（小鼠肝脏中44个脂质亚类的580个独特脂质）。结论在DIA MS/MS扫描中使用包含生物学相关脂质的纳入列表的方法与传统DDA方法一样有效，但与DDA方法相比具有更好的一致性。本研究使用Orbitrap Exploris 480进行，我们将在其他平台上进一步评估该工作流程，如果通过未来的工作进行验证，这种生物相关的离子裂解工作流程可常规用于许多研究，以提高MS/MS鉴定能力。
Introduction Data-dependent acquisition (DDA) is the most commonly used MS/MS scan method for lipidomics analysis on orbitrap-based instrument. However, MS instrument associated software decide the top N precursors for fragmentation, resulting in stochasticity of precursor selection and compromised consistency and reproducibility. We introduce a novel workflow using biologically relevant lipids to construct inclusion list for data-independent acquisition (DIA), named as BRI-DIA workflow.Objectives To ensure consistent coverage of biologically relevant lipids in LC-MS/MS-based lipidomics analysis.Methods Biologically relevant ion list was constructed based on LIPID MAPS and lipidome atlas in MS-DIAL 4. Lipids were extracted from mouse tissues and used to assess different MS/MS scan workflow (DDA, BRI-DIA, and hybrid mode) on LC-Orbitrap Exploris 480 mass spectrometer.Results DDA resulted in more MS/MS events, but the total number of unique lipids identified by three methods (DDA, BRI-DIA, and hybrid MS/MS scan mode) is comparable (580 unique lipids across 44 lipid subclasses in mouse liver). Major cardiolipin molecular species were identified by data generated using BRI-DIA and hybrid methods and allowed calculation of cardiolipin compositions, while identification of the most abundant cardiolipin CL72:8 was missing in data generated using DDA method, leading to wrong calculation of cardiolipin composition.Conclusion The method of using inclusion list comprised of biologically relevant lipids in DIA MS/MS scan is as efficient as traditional DDA method in profiling lipids, but offers better consistency of lipid identification, compared to DDA method. This study was performed using Orbitrap Exploris 480, and we will further evaluate this workflow on other platforms, and if verified by future work, this biologically relevant ion fragmentation workflow could be routinely used in many studies to improve MS/MS identification capacities.