Microchip analysis of plant glucosinolates
Microchip analysis of plant glucosinolates
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DOI：
10.1002/elps.200700635
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发表时间：
2008-06-01
期刊：
影响因子：
2.9
通讯作者：
Baba, Yoshinobu
中科院分区：
文献类型：
--
作者：
Fouad, Maged;Jabasini, Mohammad;Baba, Yoshinobu
本文描述了一种新的、选择性的分析方法，用于植物硫代葡萄糖苷的分离和定量。新方法，它利用微芯片CE（μ-CE）与荧光检测，规避了传统方法的多步骤程序特性。芥子油苷与氧杂蒽染料荧光桃红-B和曙红-B形成电荷转移复合物。葡萄糖酸盐-荧光桃红-B复合物不能在470 nm处激发。因此，复合物形成后荧光桃红-B峰强度的降低可用于定量测量拟南芥种子中的总硫代葡萄糖甙。对于定性分析，使用与曙红-B的复合物形成。在激发/发射470 nm/540 nm处检测嗜酸性粒细胞-B的灵敏度低，然而，在与黑芥子苷形成复合物（>= 3 μ g/mL）后，灵敏度增加。一个批学习，自组织地图被应用到可视化和组织分析数据到2-D矩阵与相似和相关的数据聚集在一起或彼此接近。这种有组织的矩阵用于优化分析的电泳条件。这项研究表明，在植物代谢组学分析的μ CE的潜在应用，而不使用标记荧光团。
We describe a new and selective analytical method for the separation and quantitation of plant glucosinolates. The new method, which utilizes microchip CE (mu-CE) with fluorescence detection, circumvents the multistep procedures characteristic of conventional methods. Glucosinolates form charge transfer complexes with the xanthene dyes phloxine-B and eosin-B. The glucosinotates-phloxine-B complex cannot be excited at 470 nm. Thus, the decrease in peak intensity of phloxine-B after complex formation is used to quantitatively measure total glucosinolates in Arabidopsis thaliana seeds. For qualitative analysis, complex formation with eosin-B is used. The sensitivity of eosin-B detection at excitation/emission 470 nm/540 nm was low However, sensitivity increased following complex formation with sinigrin (>= 3 mu g/mL). A batch-learning, self-organizing map was applied to visualize and organize analytical data into 2-D matrix with similar and related data clustered together or near each other. This organized matrix was used to optimize electrophoretic conditions for the analysis. This study suggests potential applications of mu-CE in plant metabolomics analyses without use of labeling fluorophores.