摘要
目的:对辽东楤木叶总皂苷进行HPLC分析并确定不同的极性部位,利用C18反相硅胶(ODS)柱分离获得不同极性部位,比较不同部位的体外抗肿瘤作用效果。方法:辽东楤木叶经70%乙醇提取后通过AB-8型大孔树脂柱,获得总皂苷,用不同体积分数甲醇经ODS柱洗脱分离获得经HPLC确定的不同部位。利用噻唑蓝(MTT)比色法测试不同部位对人肺腺癌细胞株(A549),人宫颈癌细胞株(He La)和人结肠癌细胞株(HT-29)的体外生长抑制作用。结果:由总皂苷HPLC的保留时间确定了5个极性部位,分别为A(保留时间20 min以前的部位,ODS柱上0~25%甲醇洗脱组分),B(保留时间20~45 min,ODS柱上30%~45%甲醇洗脱组分),C(保留时间50~75 min,ODS柱上50%~65%甲醇洗脱组分),D(保留时间80~110 min,ODS柱上70%~75%甲醇洗脱组分)和E(保留时间>110 min,ODS柱上80%~100%甲醇洗脱组分)。5个部位给药48 h后对3种细胞的抑制作用明显不同,A部位对3种癌细胞均无抑制作用;B部位对3种癌细胞抑制作用微弱;C,D,E部位对3种癌细胞表现出较强的抑制作用,且明显强于总皂苷,最佳的D部位对A549,Hella和HT-29抑制作用的半抑制浓度分别为4.06,4.29,3.60 mg·L^(-1)。结论:辽东楤木叶总皂苷经HPLC分析后,利用ODS柱进行分离,可获得其抗肿瘤作用的有效部位及最佳部位。
Objective： To analyze the total saponins in Aralia elata leaves by high performance liquid chromatography （HPLC） and determine the different polar sites, and separate different polar sites by C18 reversed- phase silica gel （ODS） column, then to compare the antitumor effect in vitro of different parts. Method： The total saponins was gained after extraction of A. elata leaves with 70% ethanol, and the extract was purified by AB-8 macroporous resin. Different polarity parts was determined by HPLC, and was eluted by methanol with different concentration on ODS column. Inhibitory effect of different parts on cancer ceils was determined by thiazolyl blue （MTT） colorimetry, the cancer cells included human lung adenocarcinoma cell line （A549）, human cervical cancer cell line （HeLa） and human colon cancer cell line （HT-29）. Result： Five polar sites was identified by retention time of HPLC of the total saponins. The corresponding five parts were separated by ODS column, they were 0-20 min part of HPLC and eluents with 0-25% methanol （A for short）, 20-45 min part of HPLC andeluents with 30%-45% methanol （B for short） , 50-75 min part of HPLC and eluents with 50%-65% methanol （ C for short）, 80-110 min part of HPLC and eluents with 70% -75% methanol （ D for short）, after 110 min part of HPLC and eluents with 80%-100% methanol （E for short） , respectively. The inhibitory effects of A, B, C, D and E on three kinds of ceils were significantly different. A had no effect on the three kinds of cancer cells, Inhibitory effect of B on the three kinds of cancer ceils was weak. C had strong inhibitory effects on the three kinds of cancer cells; E had stronger inhibitory effects on the three kinds of cancer cells; D showed the strongest inhibitory effect on the three kinds of cancer cells, and Its IC50 of inhibition on A549, Hella and HT-29 were 4.06, 4.29, 3.60 mg·L-1 ,respectively. Conclusion： After total saponins in A. elata leaves being identified by HPLC and separated on ODS col
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2017年第21期13-17,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
中国博士后科学基金项目(2014M551289)
国家“重大新药创制”科技重大专项(2011ZX09102-001-18)
关键词
辽东楤木叶
总皂苷
反相硅胶柱
抗肿瘤
噻唑蓝比色法
谱效关系
Aralia elata leaves
total saponins
reversed-phase silica gel column
antitumor
MTTcolorimetry method
spectrum and efficiency
