Rhizoma corydalis and Radix Angelicae Dahurica (Yuanhu-Baizhi) herbal medicine pair has been used for thousands of years and has been reported to be potentially active in recent cancer therapy. But the exact active components or fractions remain unclear. In this study, a new comprehensive two-dimensional (2D) 3-aminopropyltriethoxysilane (APTES)-decorated MCF7-cell membrane chromatography (CMC)/capcell-C18 column/time-of-flight mass spectrometry system was established for screening potential active components and clarifying the active fraction of Yuanhu-Baizhi pair. APTES was modified on the surface of silica, which can provide an amino group to covalently link cell membrane fragments with the help of glutaraldehyde in order to improve the stability and column life span of the MCF7 CMC column. The comprehensive 2D MCF7-CMC system showed good separation and identification abilities. Our screen results showed that the retention components are mainly from the alkaloids in Yuanhu (12 compounds) and the coumarins (10 compounds) in Baizhi, revealing the active fractions of Yuanhu-Baizhi herbal medicine pair. Oxoglaucine, protopine, berberine, osthole, isopimpinellin and palmitic acid were selected as typical components to test the effects on cell proliferation and their IC50 were calculated as 38.17 μM, 29.45 μM, 45.42 μM, 132.7 μM, 156.8 μM and 90.5 μM respectively. Cell apoptosis assay showed that the drug efficacy was obtained mainly through inducing cell apoptosis. Furthermore, a synergistic assay results demonstrated that oxoglaucine (representative of alkaloids from Yuanhu) and isopimpinellin (representative of coumarins from Baizhi) showed significant synergistic efficacy with GFT, indicating that these components may act on other membrane receptors. The proposed 2D CMC system could also be equipped with other cells for further applications. Besides, the follow-up in-vitro experimental strategy using cell proliferation assay, cell apoptosis assay and synergistic assay proved to be a practical way to confirm the active fractions of herbal medicine.

中文翻译：

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综合二维 APTES 修饰的 MCF7 细胞膜层析系统表征元湖-白芷中潜在的抗乳腺癌成分

延胡索和白芷（元胡-白芷）草药对已经使用了数千年，据报道在最近的癌症治疗中具有潜在的活性。但确切的活性成分或分数仍不清楚。在这项研究中，建立了一种新型的综合二维（2D）3-氨基丙基三乙氧基硅烷（APTES）装饰的MCF7细胞膜色谱（CMC）/capcell-C18柱/飞行时间质谱系统，用于筛选潜在的活性成分并阐明元胡-白芷对的活性分数。APTES在硅胶表面进行了修饰，可以提供一个氨基，在戊二醛的帮助下共价连接细胞膜片段，从而提高MCF7 CMC柱的稳定性和柱寿命。综合二维 MCF7-CMC 系统显示出良好的分离和识别能力。我们的筛选结果显示，保留成分主要来自元胡中的生物碱（12 种化合物）和白芷中的香豆素（10 种化合物），揭示了元胡-白芷药对的活性组分。选择氧代青霉素、原品碱、黄连素、蛇床子素、异青霉素和棕榈酸作为典型成分测试对细胞增殖的影响，计算其IC50分别为38.17 μM、29.45 μM、45.42 μM、132.7 μM、156.8 μM和90.5 μM。细胞凋亡试验表明，药效主要是通过诱导细胞凋亡获得的。此外，协同测定结果表明氧代青霉素（代表元湖生物碱）和异青霉素（代表白芷香豆素）与GFT有显着的协同作用，表明这些成分可能作用于其他膜受体。所提出的 2D CMC 系统还可以配备其他单元以供进一步应用。此外，使用细胞增殖测定、细胞凋亡测定和协同测定的后续体外实验策略被证明是确定草药活性组分的实用方法。