延龄草总皂苷提取纯化工艺及其抗炎活性研究  

Extraction and Purification of Total Saponins from Trillium tschonoskii and Its Anti-Inflammatory Activity

作　　者：符德静 易红[2] 郭凤倩 高慧敏[2] 李春[2] 闫利华[2] 王萍[2] 斯琴 刘晓谦 王智民[1] FU De-jing;YI Hong;GUO Feng-qian;GAO Hui-min;LI Chun;YAN Li-hua;WANG Ping;SI Qin;LIU Xiao-qian;WANG Zhi-min(Guangdong Pharmaceutical University,Guangzhou 510006,China;Institute of Chinese Materia Medica/National Engineering Laboratory of Quality Control Technology of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]广东药科大学,广东广州510006 [2]中国中医科学院中药研究所/中药质量控制技术国家工程实验室,北京100700

出　　处：《中国现代中药》2022年第10期1975-1981,共7页Modern Chinese Medicine

基　　金：国家重点研发计划项目(2017YFC1701900)。

摘　　要：目的:优选延龄草总皂苷提取纯化工艺,并对其纯化前后的抗炎活性进行对比。方法:以延龄草总皂苷提取率为考察指标,采用正交试验优化提取工艺;应用大孔吸附树脂技术,通过静、动态吸附和解吸附实验,确定最佳纯化工艺;为了证实工艺的合理性,对纯化后的总皂苷抗炎活性进行验证。结果:最佳提取工艺为乙醇体积分数75%,料液比1∶10,加热回流提取2次,每次1.5 h。纯化工艺为HPD400大孔树脂,以质量浓度为0.1 g·mL^(-1)的药液上样,上样量为折合生药(g)∶树脂(mL)=1∶2,以1 BV·h^(-1)流速上样,然后依次以水2 BV、20%乙醇水3 BV洗脱除杂,收集70%乙醇水洗脱液(5 BV),经浓缩、干燥即得延龄草总皂苷收率为7.42%,总皂苷质量分数为34.29%。该总皂苷在质量浓度为5.00、1.00μg·mL^(-1)时对脂多糖诱导RAW264.7细胞释放一氧化氮有明显的抑制作用,其半数抑制浓度(IC_(50))值为3.79μg·mL^(-1),而其醇提物的IC_(50)值为20.68μg·mL^(-1),说明精制过程实现了药效成分的富集。结论:该提取纯化工艺稳定、可行,所得到的延龄草总皂苷与其醇提物相比具有更好的抗炎活性。Objective:To optimize the extraction and purification of total saponin from Trillium tschonoskii Maxim.and compare its anti-inflammatory activities before and after purification.Methods:The extraction rate of total saponin was used as the index to optimize the extraction process by the orthogonal test.The optimized purification process was determined by static and dynamic adsorption and desorption tests by macroporous adsorption resin technology.The antiinflammatory activity of purified total saponin was tested to verify the rationality of the purification process.Results:The optimal extraction conditions were obtained as follows:ethanol volume fraction of 70%,the solid-to-liquid ratio of 1:10,and extraction twice by heating reflux,1.5 h for each time.The optimized purification process was obtained.To be specific,0.1 g·mL^(-1) sample solution was loaded on the HPD400 macroporous adsorption resin with the loading volume of the crude drug(g):resin(mL)of 1:2 at the flow rate of 1 BV·h^(-1),followed by elution and purification with 2 BV of water and 3 BV of 20%ethanol in sequence.The 70%ethanol eluate(5 BV)was collected,concentrated,and dried to obtain the total saponin of T.tschonoskii at a yield of 7.42%.The mass fraction of total saponin was 34.29%.The total saponin showed a significant inhibitory effect on LPS-induced nitric oxide(NO)release from RAW264.7 cells at mass concentrations of 5.00 and 1.00µg·mL^(-1),with an IC_(50) value of 3.79µg·mL^(-1).The IC_(50) value of its ethanol extract was 20.68µg·mL^(-1),indicating that the purification process achieved the enrichment of medicinal components.Conclusion:The extraction and purification process was stable and feasible,and the obtained total saponin of T.tschonoskii possessed superior anti-inflammatory activity to the ethanol extract.

关 键 词：延龄草 甾体皂苷 大孔树脂 抗炎活性 RAW264.7细胞 

分 类 号：R284.2[医药卫生—中药学]