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首页> 美国卫生研究院文献>other >Application of an integrated LC-UV-MS-NMR platform to the identification of secondary metabolites from cell cultures: benzophenanthridine alkaloids from elicited Eschscholzia californica (california poppy) cell cultures
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Application of an integrated LC-UV-MS-NMR platform to the identification of secondary metabolites from cell cultures: benzophenanthridine alkaloids from elicited Eschscholzia californica (california poppy) cell cultures

机译：一个集成的LC-UV-ms-NmR平台从细胞培养物中鉴定的次级代谢产物中的应用：从引发花菱草苯并菲啶生物碱（加州罂粟）的细胞培养物

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摘要

Plant cell and tissue cultures are a scalable and controllable alternative to whole plants for obtaining natural products of medical relevance. Cultures can be optimized for high yields of desired metabolites using rapid profiling assays such as HPLC. We describe an approach to establishing a rapid assay for profiling cell culture expression systems using a novel microscale LC-UV-MS-NMR platform, designed to acquire both MS and NMR each at their optimal sensitivity, by using nanosplitter MS from 4 mm analytical HPLC columns, and offline microdroplet NMR. The approach is demonstrated in the analysis of elicited Eschscholzia californica cell cultures induced with purified yeast extract to produce benzophenanthridine alkaloids. Preliminary HPLC-UV provides an overview of the changes in the production of alkaloids with time after elicitation. At the time point corresponding to the production of the most alkaloids, the integrated LC-MS-microcoil NMR platform is used for structural identification of extracted alkaloids. Eight benzophenanthridine alkaloids were identified at the sub-microgram level. This paper demonstrates the utility of the nanosplitter LC-MS/microdroplet NMR platform when establishing cell culture expression systems.
机译：植物细胞和组织培养物是用于获得医学相关性的天然产物的整株植物的可扩展和可控的替代品。可以使用诸如HPLC的快速分析测定来优化培养物的优化所需代谢物。我们描述了一种使用新型微观LC-UV-MS-NMR平台建立用于分析细胞培养表达系统的快速测定的方法，其设计用于通过使用4mm分析HPLC的纳米拔出性MS以最佳灵敏度来获取MS和NMR。列和离线Microdroplet NMR。在纯化的酵母提取物诱导的引发的Eschscholzia Californica细胞培养物分析中，证明了该方法，以产生苯蒽蒽生物碱。初步HPLC-UV概述了诱导后与时间生产生物碱的变化。在对应于制备最大生物碱的时间点时，集成的LC-MS-微膜NMR平台用于萃取的生物碱的结构鉴定。在次微小学水平上鉴定出八种苯并苯蒽醌生物碱。本文展示了纳米铂LC-MS / Microdroplet NMR平台在建立细胞培养表达系统时的效用。

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