High-speed countercurrent chromatography (HSCCC) has been successfully used for the preparative isolation of flavonoids from the ethyl acetate extracts of black currant leaves. The HSCCC separation was performed with a two-phase solvent system composed of n-hexane/EtOAc/MeOH/H2O (1:10:1:10, v/v) at a flow rate of 1.5 mL/min. When the flow rate was increased from 1.0 to 3.0 mL/min, the retention of stationary phase decreased from 60.3% to 39.7% resulting in a loss of peak resolution, while the stationary phase retention is stable with an increase in sample size from 25 to 200 mg. From 100 mg of the crude sample, HSCCC separation yielded 4.0 mg of kaempferol 3-O-galactoside, 6.0 mg of kaempferol 3-O-glucoside, and 9.0 mg of fraction I containing a mixture of hyperoside and isoquercitrin. Then, from 18 mg of fraction I, 3.0 mg of hyperoside and 11.0 mg isoquercitrin were separated by preparative HPLC by successive sample injection at every 100 min interval. Chemical structures of all these compounds were confirmed by MS and NMR.

中文翻译：

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通过高速逆流色谱和制备型 HPLC 从黑醋栗叶中分离和纯化黄酮类化合物

高速逆流色谱 (HSCCC) 已成功用于从黑醋栗叶的乙酸乙酯提取物中分离制备类黄酮。HSCCC 分离采用由正己烷/EtOAc/MeOH/H2O (1:10:1:10, v/v) 组成的两相溶剂系统，流速为 1.5 mL/min。当流速从 1.0 mL/min 增加到 3.0 mL/min 时，固定相的保留率从 60.3% 减少到 39.7%，导致峰分辨率下降，而固定相保留率随着样品量从 25% 增加到200 毫克。从 100 mg 粗样品中，HSCCC 分离产生 4.0 mg 山奈酚 3-O-半乳糖苷、6.0 mg 山奈酚 3-O-葡萄糖苷和 9.0 mg 含有金丝桃苷和异槲皮苷混合物的组分 I。然后，从 18 mg 组分 I、3.0 mg 金丝桃苷和 11。0 mg 异槲皮苷通过制备型 HPLC 以每 100 分钟的间隔连续进样进行分离。所有这些化合物的化学结构均通过 MS 和 NMR 确认。