新疆假龙胆的保肝活性部位环烯醚萜苷提取纯化工艺研究

Study on the extraction and purification of iridoids from the active sites of protecting liver in Gentianella turkestanorum

目的利用响应面法优化新疆假龙胆中保肝活性部位环烯醚萜苷的提取工艺,并优选大孔吸附树脂分离纯化新疆假龙胆中环烯醚萜苷的工艺条件。方法以新疆假龙胆中主要的3种环烯醚萜苷的提取率为考察指标,采用单因素实验结合Box-Behnken中心组合设计方法优化提取工艺,再通过考察静态和动态吸附、洗脱效果,筛选大孔吸附树脂分离纯化环烯醚萜苷的最佳工艺条件。结果新疆假龙胆环烯醚萜苷提取的最优条件为:乙醇体积分数为55%,提取时间为3.0 h,提取温度为50℃,料液比为1∶30,所得提取物环烯醚萜苷含量为52.67%(其中獐牙菜苦苷、龙胆苦苷和獐牙菜苷的含量分别为32.47%,17.41%和2.79%)。选用NKA-9型大孔吸附树脂,树脂柱径高比为1∶5进行洗脱,上样液pH值为2,质量浓度为0.5 g·mL^(-1),上样量为10 mL,吸附2 h后的树脂柱先用5 BV蒸馏水洗脱,再分别用10 BV体积分数为10%和30%的乙醇以5 BV·h^(-1)的流速洗脱。结论该方法具有较好的分离纯化效果,环烯醚萜苷类成分含量可达到59.9%。该优选工艺操作简单、稳定可行,可显著提高新疆假龙胆中环烯醚萜苷的提取率。

Objective To optimize the extraction process of iridoids from Gentianella turkestanorum by response surface methodology and to find out the suitable process conditions for the separation and purification of iridoids of Gentianella turkestanorum by using macroporous resin.Methods Based on the extraction rate of iridoids from Gentianella turkestanorum ,a single factor test and a Box-Behnken center combined design method were used to optimize the extraction process.The optimal conditions for the separation and purification of iridoid glycosides using macroporous adsorption resin were studied by investigating the static and dynamic adsorption and eluting effect.Results The optimal conditions for the extraction of iridoids from Gentianella turkestanorum were obtained as follows:ethanol volume fraction 55%,extracting time 3.0 h,extraction temperature 50 ℃,solid-liquid ratio 1∶30,iridoids extraction quantity 52.67%(the contents of swertiamarin,gentiopicroside and sweroside were 32.47%,17.41% and 2.79%).NKA-9 type macroporous adsorption resin was used with the column diameter ratio 1∶5,the sample liquid pH 2,the concentration 0.5 g·mL -1,10 mL of the sample amount,the adsorption with 5 BV first after 2 h distilled water elution,the reoccupy 10 BV 10% ethanol and 30% ethanol elution with 5 BV·h -1 velocity.Conclusion The method has a good separation and purification effect,and the total purity of iridoid glycosides can reach up to 59.9%.The optimized process operation is simple and stable,and the extraction efficiency is significantly improved.