制备型高效液相色谱法从积雪草提取物中分离纯化积雪草甙和羟基积雪草甙对照品

积雪草甙和羟基积雪草甙是积雪草及其相关制品质量控制的两个指标成分,本研究利用制备型高效液相色谱从积雪草提取物中同 时分离纯化得到这两个成分。对制备色谱的流动相组成、流速、进样量和检测波长等制备参数进行了优化。采用的色谱柱为C18柱(50 mm×200 mm,5 μm);流动相为甲醇-水(体积比为60∶40),流速100 mL/min;二极管阵列检测器在220 nm检测;进样体积为1.5 mL。在20 min的运行时间内,积雪草甙和羟基积雪草甙与干扰成分得到很好的分离,产品纯度达到98%以上。此方法具有快速高效、产品纯度高的 特点,可以用于制备积雪草甙和羟基积雪草甙对照品。

Preparation of asiaticoside and madecassoside from the extract of Centella asiatica (L.) Urb using preparative high performance liquid chromatography

A high performance liquid chromatographic (HPLC) method was successfully used to separate asiaticoside and madecassoside from the extract of Centella asiatica (L.) Urb. The raw material containing 20% asiaticoside and 45% madecassoside was dissolved in methanol and used for the preparation of the pure compounds. The flow rate of the mobile phase, injection volume and detection wavelength were optimized. The optimum operation parameters were then selected as follows: a C18 column (50 mm x 200 mm, 5 microm) with methanol-water (60 : 40, v/v) as the mobile phase at a flow rate of 100 mL/min, the detection wavelength of 220 nm, and the injection volume of 1.5 mL at the concentration of 40 mg/mL. Asiaticoside and madecassoside the can be obtained in a 20 min runtime, and the purities of the products were detected by melting point, thin layer chromatography (TLC) and HPLC method. The results showed that the purities of products were above 98%. The effective and rapid method has been successfully applied and to the preparation of asiaticoside and madecassoside from Centella asiatica (L. ) Urb.