A critical challenge in the study of botanical natural products is the difficulty of identifying multiple compounds that may contribute additively, synergistically, or antagonistically to biological activity. Herein, it is demonstrated how combining untargeted metabolomics with synergy-directed fractionation can be effective toward accomplishing this goal. To demonstrate this approach, an extract of the botanical goldenseal (Hydrastis canadensis) was fractionated and tested for its ability to enhance the antimicrobial activity of the alkaloid berberine (4) against the pathogenic bacterium Staphylococcus aureus. Bioassay data were combined with untargeted mass spectrometry-based metabolomics data sets (biochemometrics) to produce selectivity ratio (SR) plots, which visually show which extract components are most strongly associated with the biological effect. Using this approach, the new flavonoid 3,3′-dihydroxy-5,7,4′-trimethoxy-6,8-C-dimethylflavone (29) was identified, as were several flavonoids known to be active. When tested in combination with 4, 29 lowered the IC₅₀ of 4 from 132.2 ± 1.1 μM to 91.5 ± 1.1 μM. In isolation, 29 did not demonstrate antimicrobial activity. The current study highlights the importance of fractionation when utilizing metabolomics for identifying bioactive components from botanical extracts and demonstrates the power of SR plots to help merge and interpret complex biological and chemical data sets.

中文翻译：

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从植物药中鉴定增效剂和添加剂的生物化学计量学：以加拿大Hydrastis canadensis（Goldenseal）为例

植物天然产物研究中的一个关键挑战是难以鉴定多种可能对生物活性产生累加，协同或拮抗作用的化合物。在本文中，证明了将非靶向代谢组学与协同导向的分离相结合如何有效地实现这一目标。为了证明这种方法，将植物提取物（Hydrastis canadensis）的提取物进行了分级，并测试了其增强生物碱小ber碱（4）对金黄色葡萄球菌致病菌的抗菌活性的能力。。将生物测定数据与基于非靶向质谱的代谢组学数据集（生物化学计量学）结合，以生成选择性比（SR）图，该图可视地显示出哪些提取物成分与生物学效应最相关。使用这种方法，鉴定了新的类黄酮3,3'-二羟基-5,7,4'-三甲氧基-6,8- C-二甲基黄酮（29），还有几种已知的具有活性的类黄酮。当与测试4，29降低了IC ₅₀的4从132.2±1.1μM至91.5±1.1μM。孤立地，29没有显示出抗菌活性。当前的研究突出了利用代谢组学从植物提取物中鉴定生物活性成分时进行分级分离的重要性，并证明了SR图可帮助合并和解释复杂的生物和化学数据集。