决明子的化学对照品红镰霉素-6-O-β-龙胆二糖苷研究

Rubrofusarin-6-O-β-gentiobioside: chemical reference of Juemingzi(Semen Cassiae)

目的研究决明子的化学对照品红镰霉素-6-O-β-龙胆二糖苷的制备方法及其质量标准,为决明子药材及其相关产品的质量控制奠定物质基础。方法决明子药材用30%乙醇提取,采用溶剂法及硅胶、C18反相柱色谱法分离制备红镰霉素-6-O-β-龙胆二糖苷对照品。采用紫外光谱、质谱、氢谱和碳谱确认结构。采用薄层色谱法和高效液相色谱法(HPLC)进行纯度检查,薄层色谱条件为硅胶G薄层板,3种不同展开剂展开;HPLC条件为SunFire C18色谱柱(150 mm×4.6 mm,5μm),乙腈-0.1%磷酸水溶液(22∶78)为流动相,流速1.0 mL/min,于210～400 nm下检测。采用HPLC法进行均匀性检查。结果红镰霉素-6-O-β-龙胆二糖苷对照品得率0.39%,薄层色谱法检识均为单一圆整斑点;HPLC法测定纯度大于98.00%,均匀性考察结果方差分析F<F0.05(14,30),均匀性合格。结论红镰霉素-6-O-β-龙胆二糖苷对照品分离制备方法简便可行,纯度和均匀性符合化学对照品的要求,可满足决明子药材及其相关产品质量控制的需要。

Objective To study the preparation method and quality standard of chemical reference of Juemingzi （Semen Cassiae ）-rubrofusarin-6-O-β-gentiobioside, and lay a material foundation for controlling quality of crude Juemingzi and its relevant products. Methods Crude Juemingzi was extracted with 30% alcohol, and reference of rubrofusarin-6-O-β-gentiobioside was isolated and prepared by using solvent method, silica gel and C18 reversed phase column chromatography. The structure was determined by ultraviolet spectrum （UV）, mass spectrum （MS） 1H-NMR and 13C-NMR and the purity was detected by using thin layer chromatography （TLC） and high performance liquid chromatography （HPLC）. The condition of TLC was silica G thin layer plate and 3 different developing solvent systems. The procedure of HPLC was performed on SunFire C18 column （150 mm ×4.6mm,5μm）, mobile phase was acetonitrile - 0.1% of phosphoric acid solution （22： 78）, flow velocity was 0.1 mL/min, and detection wavelength was 210-400 nm. The homogeneity was checked by using HPLC. Results The yield rate of rubrofusarin-6-O-β-gentiobioside was 0.39%, and the result of TLC examination showed totally single and round spots. The purity was over 98.00% after HPLC determination, and result of variance analysis on the homogeneity showed F 〈 F0.05（14, 30） , which was appropriative. Conclusion The isolated and prepared method of rubrofusarin-6-O-β-gentiobioside is simple and feasible, and its purity and homogeneity are in aceordance with the demands of chemical reference. The method can meet the needs of the quality control of crude Juemingzi and its relevant products.