Comparative Evaluation of Data Dependent and Data Independent Acquisition Workflows Implemented on an Orbitrap Fusion for Untargeted Metabolomics
Comparative Evaluation of Data Dependent and Data Independent Acquisition Workflows Implemented on an Orbitrap Fusion for Untargeted Metabolomics
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DOI：
10.3390/metabo10040158
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发表时间：
2020-04-01
期刊：
影响因子：
4.1
通讯作者：
Fenaille, Francois
中科院分区：
文献类型：
--
作者：
Saint Hilaire, Pierre Barbier;Rousseau, Kathleen;Fenaille, Francois
Orbitrap质量分析仪在采集速度、分辨率、动态范围和灵敏度等方面的不断改进，增强了其在复杂生物基质中代谢物的大规模鉴定和定量的价值。在这里，我们报告了在高场Orbitrap Fusion(TM) Tribrid（TM）仪器上开发和优化所有理论片段离子的数据依赖采集（DDA）和顺序窗口采集（SWATH-type）数据独立采集（DIA）工作流程，用于人类血浆中代谢物的稳健鉴定和定量。通过使用47个外源分子和72个内源分子，我们比较了两种方法的效率和互补性。我们利用该质谱仪的多功能性，在优化的DDA条件下，在高、低质量分辨率和各种低能碰撞诱导解离条件下收集有意义的MS/MS光谱。我们还观察到，复杂和复合的DIA-MS/MS光谱可以有效地用于鉴定血浆中的代谢物，这要感谢由真实标准制成的参考串联光谱库，同时也为进一步鉴定未知代谢物提供了宝贵的数据资源。最后，我们发现添加多事件MS/MS采集并不会降低使用DDA和DIA工作流程的调查MS扫描的能力，以可靠地定量人血浆中代谢物至0.05 ng/mL。
Constant improvements to the Orbitrap mass analyzer, such as acquisition speed, resolution, dynamic range and sensitivity have strengthened its value for the large-scale identification and quantification of metabolites in complex biological matrices. Here, we report the development and optimization of Data Dependent Acquisition (DDA) and Sequential Window Acquisition of all THeoretical fragment ions (SWATH-type) Data Independent Acquisition (DIA) workflows on a high-field Orbitrap Fusion(TM) Tribrid(TM) instrument for the robust identification and quantification of metabolites in human plasma. By using a set of 47 exogenous and 72 endogenous molecules, we compared the efficiency and complementarity of both approaches. We exploited the versatility of this mass spectrometer to collect meaningful MS/MS spectra at both high- and low-mass resolution and various low-energy collision-induced dissociation conditions under optimized DDA conditions. We also observed that complex and composite DIA-MS/MS spectra can be efficiently exploited to identify metabolites in plasma thanks to a reference tandem spectral library made from authentic standards while also providing a valuable data resource for further identification of unknown metabolites. Finally, we found that adding multi-event MS/MS acquisition did not degrade the ability to use survey MS scans from DDA and DIA workflows for the reliable absolute quantification of metabolites down to 0.05 ng/mL in human plasma.