Fourier Transform Ion Cyclotron Resonance mass spectrometry for plant metabolite profiling and metabolite identification.
Fourier Transform Ion Cyclotron Resonance mass spectrometry for plant metabolite profiling and metabolite identification.
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DOI：
10.1007/978-1-61779-594-7_11
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发表时间：
2012-01-01
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影响因子：
--
通讯作者：
Goodacre, Royston
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文献类型：
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作者：
Allwood, J William;Parker, David;Goodacre, Royston
质谱仪(MS)通常是代谢组学研究的首选技术，在代谢组学研究中，样品材料的体积太有限，无法使用核磁共振(核磁共振)波谱。随着超高精度质谱仪的出现，如Orbitrap(分辨率10(5))和傅立叶变换离子回旋共振(FT-ICR)分析仪(分辨率可能超过10(6))，就有可能生成准确的进样样品萃取物的质量指纹图谱(通常称为轮廓，因为变量被认为是有效离散的)。在这种数据表示法中，在原始数据中检测到质量“峰”，并计算出质心质量强度。这些超高精度质谱仪的分辨率和灵敏度使得含有天然丰富的元素同位素(如(13)C、(41)K、(15)N、(17)O、(34)S和(37)Cl)的分子的代谢物信号在数据中可见。仪器的精密度如此之高，使得可以计算未知信号的高精度元素组成，从而大大有助于在通过与分析标准的比较确认未知之前选择潜在的代谢物候选对象进行注释。到目前为止，FT-ICR-MS在植物代谢组学中的应用仅限于少数几个研究，而且还没有明确的循序渐进的方法。本章为植物叶片组织的提取和FT-ICR-MS分析以及下游数据处理提供了一种严格的方法。
Mass spectrometry (MS) is usually the technique of choice for metabolomic studies where the volume of sample material is too limited for applications employing nuclear magnetic resonance (NMR) spectroscopy. With the advent of ultra-high accuracy mass spectrometers such as the Orbitrap (resolution 10(5)) and the Fourier Transform Ion Cyclotron Resonance (FT-ICR) analysers (resolution potentially in excess of 10(6)) there is the opportunity to generate an accurate mass fingerprint (often referred to as a profile since the variables are considered as effectively discrete) of an infused sample extract. In such data representations mass "peaks" are detected in the raw data and the centroid mass intensity calculated. The resolving power and sensitivity of these ultra-high accuracy mass analysers is such that metabolite signals from molecules containing naturally abundant elemental isotopes (e.g. (13)C, (41)K, (15)N, (17)O, (34)S, and (37)Cl) are visible in the data. Such is the instruments precision that it allows for the calculation of highly accurate elemental compositions for the unknown signals, thus aiding greatly in the selection of potential metabolite candidates for the annotation of unknowns prior to their confirmation by comparisons to analytical standards. The application of FT-ICR-MS to plant metabolomics has thus far been limited to a few studies and clear step-by-step methodologies are as yet unavailable. This chapter presents a rigorous method for the extraction and FT-ICR-MS analysis of plant leaf tissues as well as downstream data processing.