Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures
Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures
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DOI：
10.1002/elps.201900220
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发表时间：
2019-11-01
期刊：
影响因子：
2.9
通讯作者：
Lynn, Bert C.
中科院分区：
文献类型：
--
作者：
Kelley, Zachary D.;Rogers, D. Trent;Lynn, Bert C.
应用微流控CE*设备的CZE-MS允许快速，快速，深入分析大样本集。这种微流控CZE-MS设备，908设备ZipChip，涉及最小的样品制备，是理想的小阳离子分析物，如生物碱。本研究利用微流控装置对半边莲毛状根培养物中生物碱进行了分析。利用质谱处理软件MZmine2和统计软件Prism 8对野生型、转基因和选定突变型植物培养物的提取物进行分析和数据批量处理。通过优化电泳分离的MZmine2软件，共检测到139个特征作为基线分解峰。利用这种方法观察到初级生物碱洛比碱（C27H34N2）的相对丰度存在统计学上的显著差异，以及几种假定的“洛比碱样”分子。此外，开发并评估了一种使用微流体装置进行靶向和非靶向MS/MS实验的方法。将数据处理软件与CZE-MS数据采集相结合，可以在一个工作日内构建植物细胞培养物的全面代谢组学图谱。
Application of a microfluidic CE* device for CZE-MS allows for fast, rapid, and in-depth analysis of large sample sets. This microfluidic CZE-MS device, the 908 Devices ZipChip, involves minimal sample preparation and is ideal for small cation analytes, such as alkaloids. Here, we evaluated the microfluidic device for the analysis of alkaloids from Lobelia cardinalis hairy root cultures. Extracts from wild-type, transgenic, and selected mutant plant cultures were analyzed and data batch processed using the mass spectral processing software MZmine2 and the statistical software Prism 8. In total 139 features were detected as baseline resolved peaks via the MZmine2 software optimized for the electrophoretic separations. Statistically significant differences in the relative abundance of the primary alkaloid lobinaline (C27H34N2), along with several putative "lobinaline-like" molecules were observed utilizing this approach. Additionally, a method for performing both targeted and untargeted MS/MS experiments using the microfluidic device was developed and evaluated. Coupling data-processing software with CZE-MS data acquisition has enabled comprehensive metabolomic profiles from plant cell cultures to be constructed within a single working day.