Analysis of highly polar compounds of plant origin: Combination of hydrophilic interaction chromatography and electrospray ion trap mass spectrometry
Analysis of highly polar compounds of plant origin: Combination of hydrophilic interaction chromatography and electrospray ion trap mass spectrometry
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DOI：
10.1006/abio.2001.5513
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发表时间：
2002-02-15
影响因子：
2.9
通讯作者：
Fiehn, O
中科院分区：
文献类型：
--
作者：
Tolstikov, VV;Fiehn, O
代谢组学分析的主要目标是对生物系统中的每一种代谢物进行无偏见的相对量化。必须结合多种不同的代谢物分析技术才能做到这一点。在这里，我们报告了高极性化合物的分离和分析的概念验证研究。采用亲水作用液相色谱(HILIC)与电喷雾电离(ESI)联用对化合物进行分离分析。比较了两种类型的HILIC微孔色谱柱(聚羟乙基A和TSK凝胶酰胺80)与正常相硅胶高效液相色谱柱。用酰胺-80固定相对标样、混合物和植物样品进行了最佳分离。当与使用连续极性切换的四极离子陷阱质谱仪连接时，ESI能够检测带正电和带负电的代谢物。通过对最强烈的离子进行逐步的质谱裂解，可以识别未知化合物，然后将其包含在定制的质谱库中。利用该方法对充分展开的南瓜叶片韧皮部分泌物中的寡糖、糖苷、氨基糖、氨基酸和糖核苷酸进行了检测。采用外标法定量分析。水苏糖的检出限为0.5 ng/次(酰胺80)。根据植物的不同，所研究的韧皮部样品中水苏糖的浓度范围为1-7 mm。(C)2002年埃尔塞维尔科学公司(美国)。
The primary goal of metabolomic analysis is the unbiased relative quantification of every metabolite in a biological system. A number of different metabolite-profiling techniques must be combined to make this possible. Here we report the separation and analysis of highly polar compounds in a proof of concept study. Compounds were separated and analyzed using hydrophilic interaction liquid chromatography (HILIC) coupled to electrospray ionization (ESI) mass spectrometry. Two types of HILIC microbore columns (Polyhydroxyethyl A and TSK Gel Amide 80) were compared to normal phase silica HPLC columns. The best separations of standards mixtures and plant samples were achieved using the Amide 80 stationary phase. ESI enabled the detection of both positively and negatively charged metabolites, when coupled to a quadrupole ion trap mass spectrometer using continuous polarity switching. By stepwise mass spectrometric fragmentation of the most intense ions, unknown compounds could be identified and then included into a custom mass spectrometric library. This method was used to detect oligosaccharides, glycosides, amino sugars, amino acids, and sugar nucleotides in phloem exudates from petioles of fully expanded Cucurbita maxima leaves. Quantitative analysis was performed using external standards. The detection limit for stachyose was 0.5 ng per injection (Amide 80). The concentration of stachyose in investigated phloem samples was in the range of 1-7 mM depending on the plant. (C) 2002 Elsevier Science (USA).