College of Pharmaceutical Science, Zhejiang University of Technology, Gongda Road 1, Huzhou 313200, China.
College of Pharmaceutical Science, Zhejiang University of Technology, Gongda Road 1, Huzhou 313200, China.
J Chromatogr A. 2024 Feb 22;1717:464667. doi: 10.1016/j.chroma.2024.464667. Epub 2024 Jan 19.
In the present work, comprehensive two-dimensional reversed-phase countercurrent chromatography × reversed-phase liquid chromatography combined (2D RPCCC × RPLC) with 2D microfraction bioactive evaluation was employed to screen and isolate α-glucosidase inhibitors from Rheum palmatum L. Countercurrent chromatography was employed to improve 2D analysis and preparative separation. A selected biphasic solvent system composed of petroleum ether/ethyl acetate/methanol/water with gradient elution mode was used for the first dimension RPCCC separation (D RPCCC). Solid-phase extraction was applied to eliminate interfering polar compounds before the second dimension analysis (D RPLC). 76 components were shown in 2D contour plot in UV 280 nm. 11 Candidates were separated by a scaled-up CCC and identified by H NMR and C NMR, including anthraquinones, flavonoids, stilbenes, phenols, and glucoside derivatives. In addition, it was found that two components, resveratrol-4'-O-(6″-galloyl)glucoside (36) and lyciumaside (43) were identified as natural α-glucosidase inhibitors in Rheum palmatum L. for the first time.
在本工作中，采用综合二维反相高效逆流色谱×反相液相色谱联用（2D RPCCC×RPLC）结合二维微区生物活性评价，从大黄中筛选和分离α-葡萄糖苷酶抑制剂。反相高效逆流色谱用于改善二维分析和制备分离。选用石油醚/乙酸乙酯/甲醇/水组成的双相溶剂系统，采用梯度洗脱模式进行第一维 RPCCC 分离（D RPCCC）。在二维分析之前，采用固相萃取法（SPE）去除干扰性极性化合物（D RPLC）。在 280nm 紫外下，2D 轮廓图显示 76 个成分。通过放大 CCC 分离出 11 个候选物，并通过 1 H NMR 和 13 C NMR 进行鉴定，包括蒽醌类、黄酮类、芪类、酚类和糖苷衍生物。此外，首次发现大黄中的两种成分白藜芦醇-4'-O-(6″-没食子酰基)葡萄糖苷（36）和枸杞酰胺（43）是天然的α-葡萄糖苷酶抑制剂。
