上海同田生物技术：高速逆流色谱法分离制备丹酚酸B

Abstract: A preparative high-speed counter-current chromatography (HSCCC) was used to isolate and separate curculigoside and curculigoside B from the famous traditional Chinese medicinal herb Curculigo orchioides. Some parameters including two-phase solvent system, separation temperature,flow rate of the mobile phase and revolution speed of the apparatus were all investigated, and a successfully isolation and purification was achieved at the following conditions: the two-phase solvent system composed of ethyl acetate-ethanol-water at a volume ratio of 5:1:5(v/v/v) was selected and the lower phase of the system was used as the mobile phase at the flow rate of 2.0ml/min, and the isolation temperature and revolution speed were set at 30◦C and 800rpm.The isolation produced a total of 14.5 mg curculigoside B and 72.8 mg curculigoside with purities of 96.5% and 99.4% determined by high performance liquid chromatography (HPLC) from 300 mg crude extract after cleaning-up by D101 macroporous resin,which was necessary for the excellent purification. The recoveries of the two compounds were 91.6% and 92.5%, respectively, and the chemical structure identification was carried out by UV, MS and NMR. This separation method was more effective than some conventional techniques.

Abstract: Preparative high-speed counter-current chromatography (HSCCC), as a continuous liquid–liquid partition chromatography with no solid support matrix, combined with evaporative light scattering detection (ELSD) was employed for systematic separation and purification of non-chromophoric chemical components from Chinese medicinal herb Adenophora tetraphlla (Thunb.), Fisch. Nine compounds, includingα-spinasterol, β-sitosterol, nonacosan-10-ol, 24-methylene cycloartanol, lupenone, 3-O-palmitoyl-β-sitosterol, 3-O-β-d-glucose-β-sitosterol, eicosanoic acid and an unknown compound, were obtained. The compounds were all above 95% determined by high-performance liquid chromatography (HPLC)–ELSD, and their structures were identified by H-NMR and chemical ionization mass spectroscopy (CI-MS). The results demonstrate that HSCCC coupled with ELSD is a feasible and efficient technique for systematic isolation of non-chromophoric components from traditional medicinal herbs.

摘要：用三种方法对荷叶提取物进行分离纯化，结合薄层层析（TLC）和颜色反应，对各自分离组分跟随鉴定。结果表明：高速逆流色谱（HSCCC）分离效果好，Sephadex LH-20柱层析分离效果次之，硅胶柱层析分离效果最差；荷叶提取物经高速逆流色谱分离纯化，可以得到两种黄酮醇类单体。

摘要: 高速逆流色谱分离生物碱是目前中药有效成分分离中的热点,本文用制备型高速逆流色谱,从黄连、黄柏、十大功劳和金果榄等四种中药中分离出了几种小檗碱类生物碱.分离的基本方法都是把中药材进行粉碎,然后用工业酒精浸泡,滤液浓缩后用浓盐酸酸化过滤,滤液用氢氧化钠碱化至pH=10后用氯仿萃取,氯仿萃取液浓缩所得浸膏作为分离的样品.分离的溶剂体系主要由四氯化碳、氯仿、甲醇和稀盐酸以不同的体积比组成,分离条件是流速为2mL/min,转速为800r/min,进样量为200mg.分离所得组分用标准品进行对照,确定结构.