荷花花色苷的分离、纯化及鉴定

Seperation ,Purification and Identification of Anthocyanins Fromlotus

实验以荷花为研究对象,采用柱层析和高速逆流色谱对花色苷进行纯化分离,并利用液相色谱-质谱联用法进行花色苷鉴定。采用AB-8大孔树脂对荷花花色苷的纯化工艺进行优化,结果显示:在上样质量浓度为2 g/L、样液pH为2、上样流速为1 mL/min、上样体积为330 mL;洗脱液流速为1 m/min、pH为2、乙醇体积分数为20%的条件下,经大孔树脂纯化后,花色苷的纯度从2.12%提高到23.7%。纯化后的荷花花色苷经过高速逆流分离色谱在280 nm下分离,得到4个组分;经紫外鉴定,组分2和组分4为非花色苷类;组分1和组分3为花色苷,纯度分别为76.1%和83.3%。经液相色谱-质谱联用法检测鉴定发现,组分1含有1种花色苷,推测为锦葵色素-3-O-葡萄糖苷;组分3含有两种花色苷,推测为飞燕草素-3-O-桑布双糖苷、矢车菊-3-O-桑布双糖苷。

The anthocyanins were purified by column chromatography and high-speed countercurrent chromatography(HSCCC),and identified by liquid chromatography-mass spectrometry(LC-MS).The purification process of anthocyanin from lotus flower by AB-8 macroporous adsorption resins was optimized.The results showed that the optimal dynamic adsorption conditions were loading of 2 g/L sample solution(pH 2)at a flow rate 1 mL/min,adsorption with 330 mL,and the optimal dynamic desorption conditions were that the desorption solution was 20%ethanol(pH 2)at a flow rate of 1 mL/min.Under these conditions,the purity of anthocyanin increased from 2.12% to 23.7%.The purified anthocyanin was separated by high-speed countercurrent chromatography(HSCCC)at 280 nm,and four components were obtained;Based on the scanning of UV-Visible wavelength,components 2 and 4 were non anthocyanins,and components 1 and 3 were anthocyanins with purity of 76.1%and 83.3%.Component 1 and component 3 were identified by liquid chromatography-mass spectrometry(LC-MS).It was found that component 1 contained one anthocyanin,which was supposed to be malvacaine-3-O-glucoside and component 3 contained two anthocyanins,which were supposed to be delphinidin-3-O-sambubioside,cyanidin-3-O-sambubioside.