大孔树脂-中压柱层析联用分离纯化蓝莓花色苷

以蓝莓果实为原料,采用大孔树脂-中压柱层析联用分离纯化蓝莓花色苷。分别比较6种不同类型树脂对蓝莓花色苷静态吸附-解吸效果,优化大孔树脂分离纯化蓝莓花色苷的工艺。结果表明:D101大孔树脂对蓝莓花色苷的分离效果最佳,对花色苷的吸附属于多分子层吸附。在柱压力为1 MPa、温度25℃、上样液质量浓度为0.073 mg/m L、洗脱剂乙醇体积分数为80%、流速5 m L/min条件下,经D101大孔树脂柱分离后,花色苷纯度从5.53%增加到75.58%,提高了12.67倍。采用Sephadex LH-20中压柱层析对蓝莓花色苷进一步分离纯化,主要得到1种花色苷组分,通过高效液相色谱和高效液相色谱-电喷雾质谱联用对蓝莓花色苷进行定性和定量分析,确定该组分为矢车菊-3-O-葡萄糖苷,纯度达到90.88%。

Isolation and Purification of Anthocyanin from Blueberry by Sequential Medium Pressure Column Chromatography on Macroporous Resin and Sephadex LH-20

In this study, the anthocyanins of blueberry were isolated and purified by sequential medium pressure column chromatography using macroporous resin and Sephadex LH-20. The static adsorption and desorption of blueberry anthocyanins on six different types of resins was compared, and the separation process of anthocyanins by macroporous resin adsorption chromatography was optimized. The results showed that D101 resin gave the best separation efficiency and its adsorption behavior obeyed a multi-molecular layer adsorption. Under the following conditions: column pressure 1 MPa, temperature 25 ℃, anthocyanins concentration 0.073 mg/mL, and elution with 80% ethanol at a flow rate of 5 mL/min, the purity of anthocyanins increased by 13.67 folds from 5.53% to 75.58%. A purified anthocyanin fraction was obtained by subsequent Sephadex LH-20 column chromatography, which was identified as cyanidin-3-O-glucoside using highperformance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS) with a purity of 90.88%.