Studies On Separation And Preparation Of Anthocyanins From Berberis Kaschgarica Rupr.with High-Speed Countercurrent Chromatograhpy

The Berberis kaschgarica Rupr. is a medicinal edible plants rich in anthocyanins. Anthocyanins has important bioactivities and physiological activites, such as antioxidant,anti-mutation. anticancer, prevent platelet aggregation and improve vision. Therefore, isolate and purify anthocyanins extracted from plants has important practical significance.In this paper, The purification process of anthocyanins from berberis kaschgarica Rupr. by XAD-7HP macroporous resins was studied. Determination of the adsorption rate and desorptoin rate, filter out the best macroporous resins XAD-7HP that purified anthocyanins of Berberis kaschgarica Rupr. With the content of anthocyanins and colour value as the marker, the optimized adsorption and desorption conditions including the adsorb and desorb equilibrium time. pH. temperature, flow rate, the concentration of sample and eluent were studied. To ascertained the optimal purification:4hours of adsorption time,60%ethanol as eluant. desorption time100min, pH2.0, work temperature30℃, the concentration of anthocyanins is750mg/L. flow rate1.0mL/min. The purified anthocyanin content and color value were increased by3.15and2.21times.Selected the best solvent system by separation of four kinds of different proportions of solvent system and determination of its partition coefficient K, which is MBE-BuOH-ACN-H20-TFA=2:2:1:5:0.01. Based on this solvent system, to compare and analyze the separation of anthocyanins of Berberis kaschgarica Rupr. in different flow rate and revolution speed. The result shows that optimum conditions:a flow rate of2.0mL/min. revolution speed850rpm. the sample size50mg, volume of sample5mL. colum temperature25℃. By combination of those conditions, stationary phase retention was63.5%, retention time59.5min and good separation. Further. The effects of different flow rate and revolution speed on the stationary phase retention, resolution and retention time were studied.The anthocyanin content of Berberis kaschgarica Rupr. crude extracted for53.24mg/100g fresh fruit by HPLC. No single component in each fraction collected by HSCCC Fraction Ⅳ in peak1content up to92%, compared with the standard substance, identified as cyanidin initially.