超声提取-反相中压液相制备高纯度槐角苷的研究  

Preparation of High-purity Sophoraside by Ultrasonic Extraction-Reversed-Phase Medium-Pressure Preparative Liquid Chromatography

作　　者：王开超 莫颖宁[1] 贺灵剑 WANG Kaichao;MO Yingning;HE Lingjian(Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Jinan Dongyuan Biomedical Technology Co.,Ltd.,Jinan 250101,China)

机构地区：[1]山东中医药大学,山东济南250355 [2]济南东源生物医药技术有限公司,山东济南250101

出　　处：《山东中医杂志》2020年第8期868-873,共6页Shandong Journal of Traditional Chinese Medicine

基　　金：2018年山东省重点研发计划项目(编号:2060902);济南市科技型中小企业技术创新基金(编号:201502039)。

摘　　要：目的:以槐角为原料,建立超声提取-反相中压制备液相色谱(RP-MPPLC)从中提取纯化高纯度槐角苷的方法。方法:比较研究槐角不同部位即槐角皮和种子中槐角苷的含量差异,确定原料前处理方式;采用正交实验法优化槐角苷超声提取条件;提取物应用反相中压制备液相色谱(RP-MPPLC)纯化后,收集富含槐角苷的流分结晶处理,晶体冷冻干燥后用高效液相色谱法(HPLC)检测槐角苷纯度。结果:槐角原料的前处理方式为:槐角药材皮籽分开,取皮备用,籽做他用;确定了槐角苷的最佳提取条件为:20倍量75%乙醇超声提取2次,每次15 min;反相中压制备液相色谱纯化条件为:40%甲醇-水为流动相,流速120 mL/min,检测波长260 nm,监测波长230 nm;收集的富含槐角苷的流分浓缩至小体积,在4℃静置8 h,出现结晶,经HPLC测定其纯度为97.34%(归一化法)。结论:应用反相中压制备液相色谱可以快速高效从槐角中分离制备高纯度的槐角苷,方法简单,自动化程度高。Objective:To establish a method for the extraction and purification of high-purity Sophoroside from Huaijiao(Sophorae Fructus)by Ultrasonic Extraction-Reversed-Phase Medium-Pressure Preparative Liquid Chromatography(RP-MPPLC).Methods:The difference of sophoroside content in the peel and seed of Huaijiao was compared and studied,so as to determine the pretreatment method of the raw material.The ultrasonic extraction conditions of sophoricoside was optimized by orthogonal experiment.After purifying the extract by RP-MPPLC and collecting and crystallizing flow fractions rich in Sophoricoside,the crystals were freeze-dried to determine the purity of sophoroside by high performance liquid chromatography(HPLC)method.Results:The pretreatment method of the raw material of Huaijiao was as follow:Separating the peel and seeds of Huaijiao,then the peel was reserved,and the seeds were squeezed out for other use.The optimization result of ultrasonic extraction process was:20 times of 75%ethanol,ultrasonic extraction 2 times,15 minutes each time.The RP-MPPLC purification process was:40%methanol-water as mobile phase elution,the flow rate was 120 mL/min,the detection wavelength was 260 nm and the monitoring wavelength was 230 nm.The collected fractions rich in sophoraside were concentrated to a small volume,and left to an environment at 4℃for 8 hours,when the crystals appeared,the purity of the crystal was 97.34%(peak area normalization method).Conclusion:RP-MPPLC can be used to quickly and efficiently isolate and prepare high-purity sophoraside from Huaijiao,the method is simple and automatic.

关 键 词：槐角 槐角苷 前处理 超声波提取 反相中压制备液相色谱 高纯度 放大验证 

分 类 号：R284.2[医药卫生—中药学]