通关藤总皂苷提取工艺及抗肿瘤活性研究  

Extraction technology of total saponins from marsdenia tenacissima and its antineoplastic activity

作　　者：李媛媛[1] 贺石麟[1] 白崇智[1] 倪艳[1] 郝旭亮[1] Li Yuanyuan;He Shilin;Bai Chongzhi;Ni Yan;Hao Xuliang(Shanxi Province Institute of Traditional Chinese Medicine,Taiyuan Shanxi 030012)

机构地区：[1]山西省中医药研究院,山西太原030012

出　　处：《山西中医学院学报》2018年第5期46-49,55,共5页Journal of Shanxi College of Traditional Chinese Medicine

基　　金：山西省基础研究项目青年科技研究基金(2015021197)

摘　　要：目的:优化通关藤总皂苷的提取纯化工艺,测定通关藤总皂苷的抗肿瘤活性。方法:采用正交试验,以总皂苷收率作为指标优化通关藤总皂苷的最佳提取工艺;采用动态吸附实验,分别考察了5种不同型号的大孔树脂(D101,AB-8,ZTC-1,NKA-9,D3520)对通关藤总皂苷的吸附和解吸性能,并以吸附量、洗脱量为考察指标,对通关藤总皂苷的纯化工艺进行了筛选,以通关藤苷A为对照,采用紫外分光光度法测定通关藤总皂苷的含量;采用MTT法检测通关藤总皂苷的抗肿瘤活性。结果:通关藤总皂苷的最佳提取工艺为:加入10倍量70%乙醇,加热回流提取3次,每次2 h。ZTC-1具有最佳吸附与解析参数,最佳纯化工艺为:取通关藤粗提取物,加相当于生药量15倍的蒸馏水溶解,离心,取上清液,通过ZTC-1树脂柱,依次用水、50%乙醇洗脱至无色,弃去,再用70%乙醇洗脱1.5 BV,收集洗脱液,减压浓缩至无醇味,冷冻干燥,即得通关藤总皂苷。通关藤总皂苷对人肝癌细胞Hepg2具有抑制作用,高浓度组与空白对照组比较差异具有统计学意义(P<0.05)。当提取物浓度大于0.312 mg/mL时,人肝癌细胞Hepg2的增殖即受到抑制,且随着药物剂量的增大其差异更为显著。结论:所得提取纯化工艺简单可行,具有很好的应用前景,为通关藤总皂苷的深度开发提供依据。Objective：To optimize the extraction p arameters and determine the antineoplastic activity of saponins from marsdenia tenacissima. Methods：The best extraction parameters of total saponins from marsdenia tenacissima were investigated by orthogonal experimental design with the yield of total saponins as index. Adsorption and desorption properties of five types of macroporous adsorption resins（D101,AB-8,ZTC-1,NKA-9,D3520）to total saponins from marsdenia tenacissima were investigated by dynamic adsorption experiments,then purificating efficiency were selected with the adsorption and eluting ratio of the total saponins as index. Finally,total saponins content from marsdenia tenacissim were determined with tenacissoside A as a comparison and antineoplastic activity of sterol saponins was detected by MTT. Results：The optimum parameters were as follows∶extraction three times,2 h each time,and 10 times of 70% ethanol. The ZTC-1 provided the optimal adsorption and elution parameters,and the optimized adsorption capacity was achieved with the following condition：the crude extract was dissolved into distilled water with a ratio of 1∶15;then the resin was washed by water and50% ethanol to remove impurity and total saponins were desorbed by 1.5 BV 70% ethanol;finally,the elution was concentrated under vacuum,and dried by lyophilization. Compared with the blank control group,the cell inhibition rate of each group was significantly increased（P〈 0.05）. When the extract concentration was higher than 0.312 mg/mL,the proliferation of Hepg2 cells was inhibited,and the difference was more significant with the increase of drug dosage. Conclusion：The process of extraction and purification is simple,feasible and has a good prospect. It provides the basis for the deep development of total saponins from marsdenia tenacissima.

关 键 词：通关藤总皂苷 提取 纯化 抗肿瘤 

分 类 号：R284.2[医药卫生—中药学]