中压快速分离系统大量制备高纯度桑葚和树莓花色苷的工艺研究

Research on Large-scaled Preparation of High Purity Anthocyanins from Mulberry and Raspberry by Medium Pressure Rapid Separation System

为快速获得大量不同结构花色苷,本文以富含花色苷的桑葚和树莓为原料,通过大孔吸附树脂AB-8对两种花色苷粗提物初步分离后,利用中压快速分离系统分离得到高纯度的桑葚及树莓花色苷。制备条件为:以flash C18(80 g,20~35μm,100 A)为制备柱,两支串联,采用A相2%甲酸水,B相甲醇,流速30 mL/min,梯度洗脱程序:0~2 min,20%B;2~22 min,20%~30%B;22~32 min,30%~40%B,进样量300 mg,实现了3种不同结构花色苷的分离及纯化,桑葚中的矢车菊素-3-葡萄糖苷和矢车菊素-3-芸香糖苷产品纯度分别达到了95%和41%;树莓中的矢车菊素-3-槐糖苷和矢车菊素-3-葡萄糖苷产品纯度分别达到了60%和75%。其中桑葚中的矢车菊素-3-葡萄糖苷在32 min梯度程序内一次性可获得30 mg,且纯度为95%,可达到标准品的要求。

In order to quickly obtain a large number of different structural anthocyanins,in this paper,two crude anthocyanins were separated from rich-anthocyanins fruits namely mulberry and raspberry by macroporous adsorption resin AB-8,and high purity mulberry and raspberry anthocyanins were isolated by medium pressure rapid separation system. The specific preparation conditions were as follows:flash C18(80 g,20~35 μm,100 A)as the preparation column,two in series,using 2% of aqueous formic acid as phase A(v/v),and methanol as phase B with the gradient program of:0~2 min,20% B;2~22 min,20%~30% B;22~32 min,30%~40% B,the flow rate was 30 mL/min,and sample loading was 300 mg. The separation and purification three different structures of anthocyanins were achieved. The purity of cyanidin 3-glucoside and cyanidin 3-rutinoside,which were purified from mulberry fruits,were about 95 and 41%,respectively. Meanwhile,the purity of cyanidin 3-sophoroside,and cyanidin 3-glucoside was about 60% and 75%,respectively. Among them,30 mg of cyanidin 3-glucoside from mulberry fruits could be obtained for one run in a 32 min gradient program,and the purity was 95%,which could meet the requirements of the standard.