Metabolomics with 15N Labeling for Characterizing Missing Monoterpene Indole Alkaloids in Plants
Metabolomics with 15N Labeling for Characterizing Missing Monoterpene Indole Alkaloids in Plants
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使用 15N 标记的代谢组学来表征植物中缺失的单萜吲哚生物碱
DOI：
10.1021/acs.analchem.9b03860
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发表时间：
2020
影响因子：
7.4
通讯作者：
Saito Kazuki
中科院分区：
文献类型：
--
作者：
Nakabayashi Ryo;Mori Tetsuya;Takeda Noriko;Toyooka Kiminori;Sudo Hiroshi;Tsugawa Hiroshi;Saito Kazuki
单萜吲哚生物碱（MIA）在药用植物中具有重要的药用价值，但由于其复杂的结构，目前尚没有利用液相色谱-串联质谱（LC-MS/MS）代谢组学方法对其进行表征。我们展示了一个非靶向的代谢组学分析与15氮（N）标记，以表征MIA具有吲哚骨架的花，叶，叶柄，茎，根ofCatharanthus roseus。利用15 N-和未标记代谢物组数据的主成分分析表明，含N代谢物（N-代谢物）被15 N标记。使用保留时间、m/z值差和指示吲哚骨架的未标记产物离子atm/z 144.08的标准进行15 N-和未标记前体离子的配对。产物离子和前体离子相对于其15 N标记离子的质量位移/z值确定了其离子的N数。最后，45 MIA分子式明确确定使用所确定的N数。利用MS/MS相似性进行的生物碱网络分析显示了MIA之间的结构共性和独特性。其中，安替比林是确定使用一个真实的标准化合物。多代谢组学使用LC-MS/MS和成像质谱表明，antirhine积累相当大的表皮和维管柱的根。开发的方法表明存在缺失的MIAs。这种方法的修改将鉴定含有羟基化或甲氧基化吲哚骨架的其他MIA。
Monoterpene indole alkaloids (MIAs) in medicinal plants remain uncharacterized owing to their complicated structure by metabolomics using liquid chromatography–tandem mass spectrometry (LC–MS/MS) despite their pharmaceutical importance. We demonstrate an untargeted metabolome analysis with15nitrogen (N) labeling to characterize MIAs having an indolic skeleton in the flowers, leaves, petioles, stems, and roots ofCatharanthus roseus. Principal component analysis using15N- and nonlabeled metabolome data showed that N-containing metabolites (N-metabolites) are labeled with15N. Paring of the15N- and nonlabeled precursor ions were performed using the criteria of retention time, difference ofm/zvalue, and a nonlabeled product ion atm/z144.08 that indicates an indolic skeleton. The mass shift of them/zvalue of the product and precursor ions to their15N-labeled ions identified the number of N of their ions. Finally, molecular formula of 45 MIAs was unambiguously identified using the identified N number. The alkaloid network analysis using the MS/MS similarity showed the structural commonness and uniqueness among the MIAs. Of them, antirhine was identified using an authentic standard compound. Multimetabolomics using LC–MS/MS and imaging mass spectrometry showed that antirhine accumulates considerably in the epidermis and vascular cylinder of the roots. The developed approach showed the existence of the missing MIAs. The modification of this approach will identify other MIAs that contain a hydroxylated or methoxylated indolic skeleton.