小春花总黄酮提取纯化工艺研究

Study on Methods for Extraction and Purification of FST

目的：建立小春花总黄酮的提取、纯化工艺,为总黄酮化学成分、药理研究和制剂开发提供基础。方法：以小春花总黄酮为指标,考察提取工艺;采用大孔吸附树脂、聚酰胺树脂纯化小春花总黄酮,对树脂型号、上样浓度、上样液pH值、洗脱溶剂的用量等进行了考察,确定最佳工艺。结果：小春花总黄酮的最佳提取工艺为：6倍量70%乙醇提取3次,每次2 h;得到总黄酮的平均含量4.14 mg/g,转移率均达到90%以上;大孔吸附树脂纯化小春花总黄酮的最佳工艺为：以D101型大孔吸附树脂为吸附剂,上样液浓度p H为5、浓度为1.22 mg/mL,以流速2 mL/min过柱,用水6BV进行洗脱,再用70%乙醇4BV洗脱,得到的小春花总黄酮含量达7.25%,得率均在75%以上;聚酰胺树脂纯化最佳工艺为：以100-200型聚酰胺树脂为吸附剂,上样浓度为1.18 mg/mL,流速2mL/min过柱,再用70%乙醇4BV洗脱,得到小春花总黄酮的含量符合制剂学要求。结论：本部分建立了小春花总黄酮的提取工艺,并采用大孔吸附树脂、聚酰胺树脂进行纯化,提取的总黄酮含量≥50%,满足制剂开发的需求。

Objective： To establish the method for the extraction and purification of FST and analyze its main chemical constituents so that the quality could meet the requirements of new formulation development. Methods： The extraction method was evaluated by testing FST. They were purified by macroporous adsorption resin and polyamide resin. The resin type,loading concentration,p H value of the sample solution and the amount of elution solvent were optimized. The main components of FST were analyzed by HPLC-MS technology. Results： The optimal extraction process of FST was as follows： 6 times the amount of 70% ethanol,extraction 3 times,2 hours for each time. The average content of FST was4. 14 mg/g and the transfer rate was over 90%. The optimal purification conditions on macroporous adsorption resin： the column was eluted at a flow rate of 2 mL/min with 6 BV of water,and then with 4 BV of 70% ethanol. The obtained content of total flavonoids was 7. 25% and the yield exceeded 75%. The optimal purification conditions on polyamide resin：the column was eluted at a flow rate of 2 mL/min with 4 BV of 70% ethanol. The obtained content of FST was in accordance with the requirements for formulation preparation. Conclusion： FST extraction process was set up in present part. After purification by macroporous resin or polyamide resin,the FST met the requirements for formulation preparation.