Targeting unique biological signals on the fly to improve MS/MS coverage and identification efficiency in metabolomics.
Targeting unique biological signals on the fly to improve MS/MS coverage and identification efficiency in metabolomics.
复制标题
DOI：
10.1016/j.aca.2021.338210
复制
发表时间：
2021-03-08
影响因子：
6.2
通讯作者：
Patti GJ
中科院分区：
文献类型：
--
作者：
Cho K;Schwaiger-Haber M;Naser FJ;Stancliffe E;Sindelar M;Patti GJ
当使用液相色谱/质谱（LC/MS）来执行非靶向代谢组学时，通常从生物提取物中检测数千个特征。虽然在单次色谱运行中收集每种非嵌合MS/MS数据是不切实际的，但这通常是不必要的，因为大多数特征不对应于具有生物相关性的独特代谢物。在这里，我们表明，相对简单的数据处理策略，可以应用在飞行过程中的数据采集与Orbitrap ID-X，如空白减法和完善的加合物或同位素计算，减少功能的数量，以目标MS/MS分析的数量级为各种类型的生物基质。我们证明，注释这些非生物污染物和冗余在数据采集过程中的真实的时间，使全面的MS/MS数据被收购的每个剩余的功能在一个单一的碰撞能量。为了确保应用适当的碰撞能量，我们引入了一种方法，该方法使用Orbitrap ID-X中的一系列隐藏离子阱扫描来找到每个特征的最佳值，然后可以将其应用于后续的高分辨率Orbitrap扫描。来自100种代谢物标准品的数据表明，与单独使用单一固定碰撞能量相比，这种碰撞能量的实时优化导致信息量更大的MS/MS模式。作为评估整个工作流程的基准，我们通过独立地对E.大肠杆菌样本进行认证分析。虽然认证导致特征数量的更严格减少，但在Orbitrap ID-X上使用空白减法进行动态注释不会不适当地丢弃独特的生物代谢物。总之，我们的结果表明，当在采集期间注释特征并通过使用并行质谱检测选择碰撞能量时，可以在单个LC/MS/MS运行中获得样品中>90%的独特生物代谢物的最佳碎片化数据。
When using liquid chromatography/mass spectrometry (LC/MS) to perform untargeted metabolomics, it is common to detect thousands of features from a biological extract. Although it is impractical to collect non-chimeric MS/MS data for each in a single chromatographic run, this is generally unnecessary because most features do not correspond to unique metabolites of biological relevance. Here we show that relatively simple data-processing strategies that can be applied on the fly during acquisition of data with an Orbitrap ID-X, such as blank subtraction and well-established adduct or isotope calculations, decrease the number of features to target for MS/MS analysis by up to an order of magnitude for various types of biological matrices. We demonstrate that annotating these non-biological contaminants and redundancies in real time during data acquisition enables comprehensive MS/MS data to be acquired on each remaining feature at a single collision energy. To ensure that an appropriate collision energy is applied, we introduce a method using a series of hidden ion-trap scans in an Orbitrap ID-X to find an optimal value for each feature that can then be applied in a subsequent high-resolution Orbitrap scan. Data from 100 metabolite standards indicate that this real-time optimization of collision energies leads to more informative MS/MS patterns compared to using a single fixed collision energy alone. As a benchmark to evaluate the overall workflow, we manually annotated unique biological features by independently subjecting E. coli samples to a credentialing analysis. While credentialing led to a more rigorous reduction in feature number, on-the-fly annotation with blank subtraction on an Orbitrap ID-X did not inappropriately discard unique biological metabolites. Taken together, our results reveal that optimal fragmentation data can be obtained in a single LC/MS/MS run for >90% of the unique biological metabolites in a sample when features are annotated during acquisition and collision energies are selected by using parallel mass spectrometry detection.
登录
查看更多内容
影响因子：
4.1
作者：
Saint Hilaire, Pierre Barbier;Rousseau, Kathleen;Fenaille, Francois
通讯作者：
Fenaille, Francois
影响因子：
7.4
作者：
Mahieu NG;Patti GJ
通讯作者：
Patti GJ
影响因子：
7.4
作者：
Nikolskiy, Igor;Mahieu, Nathaniel G.;Chen, Ying-Jr;Tautenhahn, Ralf;Patti, Gary J.
通讯作者：
Patti, Gary J.
影响因子：
7.4
作者：
Stupp, Gregory S.;Clendinen, Chaevien S.;Ajredini, Ramadan;Szewc, Mark A.;Garret, Timothy;Menger, Robert F.;Yost, Richard A.;Beecher, Chris;Edison, Arthur S.
通讯作者：
Edison, Arthur S.
影响因子：
7.4
作者：
Domingo-Almenara, Xavier;Montenegro-Burke, J. Rafael;Siuzdak, Gary
通讯作者：
Siuzdak, Gary