高速逆流色谱法分离红葡萄皮中的花色苷

Isolation of anthocyanins from red grape skins by high-speed countercurrent chromatography

建立高速逆流色谱(HSCCC)法分离制备红葡萄皮中花色苷单体的方法。以乙腈-正丁醇-甲基叔丁基醚-水-三氟乙酸(1∶40∶1∶50∶0.01,V/V)为溶剂体系,上相(有机相)为固定相,下相(水相)为流动相,流速为2.0 m L/min,转速为950 r/min,进样量为200 mg,分离得到的组分利用紫外-可见(UV)光谱和液质联用(HPLC-MS)技术进行定性分析。结果表明,经过一次高速逆流分离即可得到3种花色苷单体,分别为飞燕草色素-3-O-葡萄糖苷、锦葵色素-3-O-葡萄糖苷和芍药色素-3-O-葡萄糖苷,高效液相色谱(HPLC)峰面积归一法计算其纯度分别为93.7%、95.2%、91.6%。采用高速逆流色谱法成功从红葡萄皮中一次性分离得到3种高纯度的花色苷单体,其中芍药色素-3-O-葡萄糖苷为首次分离得到。

The method for separation and preparation of anthocyanin monomers from red grape skins by high-speed countercurrent chromatography （HSCCC） was established. Using acetonitrile-n-butyl alcohol-methyl tert-butyl ether （MTBE）-water-trifluoroacetic acid （TFA） （1：40：1：50：0.01, V/V） as solvent system, upper-phase （organic phase） as stationary phase and lower phase （water phase） as mobile phase, with the flow rate of 2 ml/min, rotation speed of 950 r/min and sample injection of 200 mg, the compounds obtained were identified by ultraviolet-visible （LrV） spectroscopy and high performance liquid chromatography-mass spectrometry （HPLC-MS） for qualitative analysis. The results showed that the major anthocyanins were delphinidin-3-O-glucoside, malvidin-3-O-glucoside and peonidin-3-O-glucoside. They were successfully separated one from another through HSCCC with the purity of 93.7%, 95.2% and 91.6%, respectively. Three anthocyanins monomers with high purity were successfully one-time separated from red grape skins by HSCCC and the compound ofpeonidin-3-O-glucoside was firstly isolated in this work.