A rapid and efficient method for the separation and purification of ombuoside from Gynostemma pentaphyllum by microwave-assisted extraction coupled with high-speed counter-current chromatography (HSCCC) was successfully developed. Using an orthogonal array design L9 (34), the extraction conditions, including microwave power, irradiation time, solid-to-liquid ratio and extraction times, were optimized. Ombuoside was isolated and purified from the crude extraction by HSCCC with two-phase solvent system composed of n-hexane:ethyl acetate:ethanol:water (5:6:5:5, v/v) in a single run. A 210 mg quantity of the crude extract containing 2.16% ombuoside was loaded, yielding 3.9 mg of ombuoside at 96.7% purity. The chemical structure of ombuoside was determined by comparison with the high-performance liquid chromatography retention time of standard substance as well as UV, FT-IR, ESI-MS, 1H NMR and 13C NMR spectra. The purified ombuoside had strong 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radical scavenging activities.

中文翻译：

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微波辅助萃取-高速逆流色谱法分离和纯化绞股蓝中的翁布苷。

成功开发了一种快速，高效的微波辅助萃取-高速逆流色谱分离纯化绞股蓝总皂苷的方法。使用正交阵列设计L9（34），优化了提取条件，包括微波功率，辐照时间，固液比和提取时间。从HSCCC粗提物中分离出Ombuoside，并将其纯化，一次使用由正己烷：乙酸乙酯：乙醇：水（5：6：5：5，v / v）组成的两相溶剂系统。装载210mg量的含有2.16％的核苷的粗提物，得到3.9mg纯度为96.7％的核苷。通过与标准物质的高效液相色谱保留时间以及UV，FT-IR，ESI-MS，1H NMR和13C NMR光谱比较，确定了翁布赛苷的化学结构。纯化的翁布甙具有很强的1,1-二苯基-2-吡啶并肼基和羟基自由基清除活性。