毛花猕猴桃叶中AE的含量测定方法建立及提取纯化工艺研究(1)
[摘要]目的 建立毛花獼猴桃叶中2α，3α，24-三羟基-12-烯-28-乌苏酸(AE)的检测方法并考察提取纯化工艺。方法 以AE为对照品，利用高效液相色谱(HPLC)结合蒸发光散射检测器(ELSD)，建立AE的含量测定方法;采用单因素及正交试验，考察最佳提取工艺;采用单因素实验，考察最佳纯化工艺。结果 建立了稳定可行的AE含量测定方法，并测定了5个不同产地的猕猴桃叶中AE的含量，发现不同产地之间含量差异较大;考察得出最佳提取工艺为：用70%乙醇提取2次，第1次15倍量提取2 h，第2次10倍量提取1 h;最佳纯化工艺为：将提取液浓缩至0.50 g生药/ml，浓缩液静置8 h后，弃去上清液，取沉淀，加入30%乙醇至浓度为0.50 g生药/ml，充分搅拌，再静置8 h，弃去上清液，取沉淀，加入适量石油醚(60-90)充分洗涤。最终得到的AE中间体平均含量为36.25%，从药材到中间体的得率也达到55.96%。结论 本实验建立的含量测定方法稳定可行，可用于猕猴桃叶中AE的含量测定;优选出的工艺操作简单可行、重复性好，可推广于工业化大生产。
[关键词]毛花猕猴桃叶;2α，3α，24-三羟基-12-烯-28-乌苏酸;含量测定;提取工艺;纯化工艺
[中图分类号] R917 [文献标识码] A [文章编号] 1674-4721(2020)9(a)-0018-05
[Abstract] Objective To establish the method for determination of 2α，3α，24-trihydroxyurs-12-en-28-oic acid (AE) from the leaves of Actinidia eriantha Benth.， and optimize the extraction and purification process. Methods The content determination method of AE was established by high performance liquid chromatography-evaporative light-scattering detector (HPLC-ELSD) using AE as reference substance. The optimal extraction process was researched by single factor test and orthogonal test. The optimal purification process was researched by single factor test. Results The stable content determination method of AE was established， and it was found that the content of AE in Actinidia eriantha Benth. leaves from 5 different habitats was quite different. The optimal process of the extraction was as follows： extraction for 2 hours with 15-fold 70% ethanol in the first time， and extraction for 1 hour with 10-fold 70% ethanol in the second time. The optimal purification process was as follows： concentrating the extract to 0.50 g crude drug/ml， discarding the supernatant after sit for 8 hours， adding 30% ethanol into the sediment to let the concentration become 0.50 g crude drug/ml， and stirring thoroughly， discarding the supernatant after sit for 8 hours again， adding the appropriate amount of petroleum ether (60-90) into the sediment， and washing thoroughly. The average content of AE intermediate was 36.25%， and the yield from medicinal materials to intermediates also reached 55.96%. Conclusion The method established in this experiment is stable and feasible， and can be used for the determination of AE in Actinidia eriantha Benth. leaves， the optimized process is simple， stable and repeatable， which is suitable for industrial production.
[Key words] Leaves of Actinidia eriantha Benth.; 2α，3α，24-trihydroxyurs-12-en-28-oic acid; Content determination; Extraction process; Purification process, 百拇医药(王永红 王雪莹 彭世陆 禹奇男 李红亮)
[关键词]毛花猕猴桃叶;2α，3α，24-三羟基-12-烯-28-乌苏酸;含量测定;提取工艺;纯化工艺
[中图分类号] R917 [文献标识码] A [文章编号] 1674-4721(2020)9(a)-0018-05
[Abstract] Objective To establish the method for determination of 2α，3α，24-trihydroxyurs-12-en-28-oic acid (AE) from the leaves of Actinidia eriantha Benth.， and optimize the extraction and purification process. Methods The content determination method of AE was established by high performance liquid chromatography-evaporative light-scattering detector (HPLC-ELSD) using AE as reference substance. The optimal extraction process was researched by single factor test and orthogonal test. The optimal purification process was researched by single factor test. Results The stable content determination method of AE was established， and it was found that the content of AE in Actinidia eriantha Benth. leaves from 5 different habitats was quite different. The optimal process of the extraction was as follows： extraction for 2 hours with 15-fold 70% ethanol in the first time， and extraction for 1 hour with 10-fold 70% ethanol in the second time. The optimal purification process was as follows： concentrating the extract to 0.50 g crude drug/ml， discarding the supernatant after sit for 8 hours， adding 30% ethanol into the sediment to let the concentration become 0.50 g crude drug/ml， and stirring thoroughly， discarding the supernatant after sit for 8 hours again， adding the appropriate amount of petroleum ether (60-90) into the sediment， and washing thoroughly. The average content of AE intermediate was 36.25%， and the yield from medicinal materials to intermediates also reached 55.96%. Conclusion The method established in this experiment is stable and feasible， and can be used for the determination of AE in Actinidia eriantha Benth. leaves， the optimized process is simple， stable and repeatable， which is suitable for industrial production.
[Key words] Leaves of Actinidia eriantha Benth.; 2α，3α，24-trihydroxyurs-12-en-28-oic acid; Content determination; Extraction process; Purification process, 百拇医药(王永红 王雪莹 彭世陆 禹奇男 李红亮)