草豆蔻有效成分的半制备型高效液相色谱分离

目的]建立从草豆蔻中分离纯化4种主要有效成分半制备型高效液相色谱法。方法]使用C18柱(250 mm×25.4 mm I.D.,10μm),考察了流动相的组成、流速和上样量对分离效果的影响,确定适宜的色谱条件;按制备型色谱条件进行洗脱,根据检测信号在线收集产品流出液,分离所得的馏分经HPLC法进行纯度检测。结果]适宜的色谱条件是:流动相为甲醇-水梯度洗脱,流速为25 m L/min,上样量为232 mg,检测波长为300 nm。经过一次分离即可得到4种成分,经紫外光谱和核磁共振波谱鉴定4种成分分别为山姜素、松属素、小豆蔻明和桤木酮,经高效液相色谱检测纯度分别为98.7%、99.3%、98.4%和99.1%。结论]与传统的分离方法相比,该方法具有高效、简便、快速的特点,适用于草豆蔻中主要有效成分的快速制备。

Separation of Active Components in Alpinia katsumadai Hayata by Semi-preparative High Performance Liquid Chromatography

Objective] The aim was to develop a semi-preparative high performance liquid chromatography (SP-HPLC) method for separation and purification of four effective components from Alpinia katsumadai Hayata.Method] Using C18 column (250 mm ×25.4 mm I.D.,10 μm), the optimum separation conditions were obtained through investigating the effects caused by the composition and flow rate of the mobile phase and the sample size.Elution was performed by preparative chromatography,product effluent was collected online according to detection signal,the pu-rity of separated fraction was detected by HPLC method. Result] The suitable conditions were listed as follows:the mobile phase was methanol-water in stepwise elution mode,the flow rate of the mobile phase was 25 mL/min,the sample size was 232 mg,and the detection wavelength was 300 nm.Under the above conditions,four compounds obtained purification through one-step separation.They were identified as alpinetin,pino-cembrin,cardamonin and alnustone by ultraviolet spectrum (UV) and nuclear magnetic resonance (NMR) analysis,and the purities of the four compounds were 98.7%,99.3%,98.4%and 99.1%as determined by high performance liquid chromatography (HPLC).Conclusion] Compa-ring with the traditional separation method,the SP-HPLC method developed in the paper has many advantages such as high efficiency,facility,and high speed.And it was suitable for rapid preparation of effective components from A.katsumadai.