Improving global feature detectabilities through scan range splitting for untargeted metabolomics by high-performance liquid chromatography-Orbitrap mass spectrometry
Improving global feature detectabilities through scan range splitting for untargeted metabolomics by high-performance liquid chromatography-Orbitrap mass spectrometry
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DOI：
10.1016/j.aca.2016.05.017
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发表时间：
2016-08-03
影响因子：
6.2
通讯作者：
Huber, Christian G.
中科院分区：
文献类型：
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作者：
Ranninger, Christina;Schmidt, Lukas E.;Huber, Christian G.
非靶向代谢组学旨在获得生物样品中存在的尽可能多的低分子生物分子的定量信息。在非靶向全球规模研究中，通过高效液相色谱-质谱法（HPLC-MS），质谱采集参数的微小变化可能对代谢物的可检测性产生显著影响。使用人肾近端小管细胞的全细胞裂解物，我们提出了一个系统的全球规模的研究质谱扫描参数和采集后数据处理的影响，在全细胞裂解物中可检测到的代谢物的数量和强度。离子传输和离子收集效率在基于Orbitrap的质谱仪基本上取决于m/z范围扫描，其中，理想情况下，对于所研究的各个质量范围，需要不同的仪器设置。因此，我们将与代谢物相关的m/z 50-1000的完整扫描范围分为两个单独的部分（m/z 50-200和m/z 200- 1，000），允许独立调整两个质量范围的离子传输参数。严格评估了三种不同的实施方式，包括在单次扫描中从m/z 50-1000进行扫描，或在两次交替扫描中从m/z 50-200和从m/z 200-1000进行扫描，或在m/z 50-200和m/z 200-1000扫描范围内进行两次单独的HPLC-MS运行。检测到的特征进行严格的背景过滤和质量控制，以获得可靠的代谢物特征，为后续的差分quantization.The最有效的方法在特征数量，这构成了统计分析，识别的基础，并为产生生物学假设，是两个不同的质量范围的单独分析。与涉及单个扫描范围的分析相比，这导致可检测代谢物特征的数量增加，特别是在较高质量范围（m/z大于400）中，增加了2.5倍（阴性模式）至6倍（阳性模式）。在正离子模式下，可确信检测的特征总数为560，在负离子模式下为436。(C)© 2016 Elsevier B. V.版权所有。
Untargeted metabolomics aims at obtaining quantitative information on the highest possible number of low-molecular biomolecules present in a biological sample. Rather small changes in mass spectrometric spectrum acquisition parameters may have a significant influence on the detectabilities of metabolites in untargeted global-scale studies by means of high-performance liquid chromatography-mass spectrometry (HPLC-MS). Employing whole cell lysates of human renal proximal tubule cells, we present a systematic global-scale study of the influence of mass spectrometric scan parameters and post-acquisition data treatment on the number and intensity of metabolites detectable in whole cell lysates.Ion transmission and ion collection efficiencies in an Orbitrap-based mass spectrometer basically depend on the m/z range scanned, which, ideally, requires different instrument settings for the respective mass ranges investigated. Therefore, we split a full scan range of m/z 50-1000 relevant for metabolites into two separate segments (m/z 50-200 and m/z 200-1,000), allowing an independent tuning of the ion transmission parameters for both mass ranges. Three different implementations, involving either scanning from m/z 50-1000 in a single scan, or scanning from m/z 50-200 and from m/z 200-1000 in two alternating scans, or performing two separate HPLC-MS runs with m/z 50-200 and m/z 200-1000 scan ranges were critically assessed. The detected features were subjected to rigorous background filtering and quality control in order to obtain reliable metabolite features for subsequent differential quantification.The most efficient approach in terms of feature number, which forms the basis for statistical analysis, identification, and for generating biological hypotheses, was the separate analysis of two different mass ranges. This lead to an increase in the number of detectable metabolite features, especially in the higher mass range (m/z greater than 400), by 2.5 (negative mode) to 6-fold (positive mode) as compared to analysis involving a single scan range. The total number of features confidently detectable was 560 in positive ion mode, and 436 in negative ion mode. (C) 2016 Elsevier B.V. All rights reserved.