Nontargeted metabolome analysis by use of fourier transform ion cyclotron mass spectrometry
Nontargeted metabolome analysis by use of fourier transform ion cyclotron mass spectrometry
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DOI：
10.1089/15362310260256882
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发表时间：
2002-01-01
期刊：
影响因子：
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通讯作者：
Goodenowe, Dayan B.
中科院分区：
文献类型：
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作者：
Aharoni, Asaph;De Vos, C. H. Ric;Goodenowe, Dayan B.
先进的功能基因组工具现在允许基因和蛋白质表达的平行和高通量分析。尽管这些信息对我们理解基因功能至关重要，但它对与代谢相关的表型变化提供的认识不足。在这里，我们介绍了一种基于高容量傅立叶变换离子回旋质谱（FTMS）的方法，能够进行非靶向代谢分析，适用于大量生物样品（例如植物突变群体）的相似性和差异性的快速筛选。代谢物的分离完全通过超高质量分辨率实现；根据准确的质量测定，通过测定代谢物的元素组成来鉴定其所属的推定代谢物或代谢物类别；通过内部校准比较各质量的绝对强度来实现相对定量。通过直接（连续流）注射引入植物粗提取物，并通过电喷雾电离（ESI）或常压化学电离（APCI）在正或负电离模式下进行电离。我们首先分析了草莓果实发育的四个连续阶段，并确定了与已知果实代谢物相对应的大范围质量水平的变化。这些数据还揭示了从未成熟水果到成熟水果的代谢转变的新信息。在另一组实验中，该方法被用于跟踪烟草花代谢谱的变化，草莓MYB转录因子过表达和花瓣颜色改变。转基因植株与对照植株只有9个质量块存在差异，其中主要花色素花青素-3-鼠李糖苷对应的质量块存在差异。结果证明了FTMS方法用于非靶向和快速代谢“指纹”的可行性和实用性，这将极大地加快当前研究代谢组的努力，并在任何生物系统中推导基因功能。
Advanced functional genomic tools now allow the parallel and high-throughput analyses of gene and protein expression. Although this information is crucial to our understanding of gene function, it offers insufficient insight into phenotypic changes associated with metabolism. Here we introduce a high-capacity Fourier Transform Ion Cyclotron Mass Spectrometry (FTMS)-based method, capable of nontargeted metabolic analysis and suitable for rapid screening of similarities and dissimilarities in large collections of biological samples (e.g., plant mutant populations). Separation of the metabolites was achieved solely by ultra-high mass resolution; Identification of the putative metabolite or class of metabolites to which it belongs was achieved by determining the elemental composition of the metabolite based upon the accurate mass determination; and relative quantitation was achieved by comparing the absolute intensities of each mass using internal calibration. Crude plant extracts were introduced via direct (continuous flow) injection and ionized by either electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) in both positive or negative ionization modes. We first analyzed four consecutive stages of strawberry fruit development and identified changes in the levels of a large range of masses corresponding to known fruit metabolites. The data also revealed novel information on the metabolic transition from immature to ripe fruit. In another set of experiments, the method was used to track changes in metabolic profiles of tobacco flowers overexpressing a strawberry MYB transcription factor and altered in petal color. Only nine masses appeared different between transgenic and control plants, among which was the mass corresponding to cyanidin-3-rhamnoglucoside, the main flower pigment. The results demonstrate the feasibility and utility of the FTMS approach for a nontargeted and rapid metabolic "fingerprinting," which will greatly speed up current efforts to study the metabolome and derive gene function in any biological system.