环糊精键合凝胶介质分离纯化金银花中绿原酸

Purification of Chlorogenic Acid from Honeysuckle by Oligo-β-cyclodextrin Coupled Sepharose HP Column

采用寡聚环糊精键合凝胶柱分离纯化金银花中的绿原酸,考察了流动相、柱温、负载量及介质再生等因素对分离纯化效果的影响.分别用脲和十二烷基磺酸钠掩盖氢键和疏水作用,探讨了分离机理.结果表明,以水:乙醇:醋酸体积比为85:10:5的混合溶液,负载量为0.5mg/mL进行等度洗脱,常温下通过一次色谱即可分离纯化绿原酸,其收率和纯度分别达到65%和97%以上.介质使用10次后需要再生,先后经0.3mol/LNaOH、水和30%乙酸简单再生即可重复利用.

An epichlorohydrin polymerized β-cyclodextrin ligand coupled to brominated allyl-group substituted Sepharose HP was used for one-step purification of chlorogenic acid from honeysuckle. The experimental conditions, such as mobile phase, temperature, loading capacity, on the separation were optimized. The separation conditions were optimized. The optimized mobile phase was H2O-ethanolacetic acid （85：10：5） with the flow rate of 1.0 mL/min. The loading capacity of the wet gel was 0.5 mg/mL （wet gel） for samples. The effect of temperature was discussed, the mechanism of separation was confirmed by adding urea and SDS respectively into the mobile phase. Adopting the optimum mobile phase, a chlorogenic acid purity of about 97% with a recovery rate of approximate 65% could be achieved by proper peak cutting. The result was affirmed by MS spectra. The column could be reused for 10 times with good separation. Full separation efficiency could be restored by cleaning the column with 0.3 mol/L NaOH followed by distilled water and 30% acetic acid.