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目的:以冬凌草甲素,毛叶香茶菜素E,毛栲利素,香茶菜素B为指标,建立HPLC同时测定鄂西香茶菜中4个二萜的质量控制方法,并对4个成分进行体外抗肿瘤活性的评价。方法:采用YMC C18反相色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-水(38∶62),流速1.0 m L·min~(-1),柱温30℃,检测波长230 nm;采用噻唑蓝(MTT)比色法测定了鄂西香茶菜中4个指标性成分对HCT~(-1)16,Hep G2,BGC-823,MB-231,A2780细胞的增殖抑制作用。结果:冬凌草甲素,毛叶香茶菜素E,毛栲利素,香茶菜素B分别在0.085 6~0.856,0.014 04~0.140 4,0.070 8~0.708,0.018 98~0.189 8 g·L~(-1)呈良好的线性关系,平均回收率分别为101.78%,101.70%,100.23%,104.60%。4个二萜均显示了一定的细胞增殖抑制作用,其中冬凌草甲素,毛栲利素对5种细胞株的增值抑制率均>50%。冬凌草甲素、毛栲利素对5株肿瘤细胞株的细胞毒作用均优于毛叶香茶菜素E和香茶菜素B。半数抑制浓度(IC50)分别为4.10~6.46,2.34~4.77μmol·L~(-1)。结论:该方法简便,灵敏,重复性好,可用于鄂西香茶菜中主要细胞增殖抑制二萜的质量控制。
OBJECTIVE: To establish a method for the simultaneous determination of 4 diterpenes in oriental tea in Hubei Province by using Rubescens A, Erigeron, Erigeron and Cinnamon B as indexes The four components in vitro antitumor activity evaluation. Methods: A mobile phase of acetonitrile-water (38:62) was used at a flow rate of 1.0 mL · L -1 on a YMC C18 reversed-phase column (4.6 mm × 250 mm, 5 μm) nm. The inhibitory effects of four index components of Xiang tea from western Hubei province on the proliferation of HCT-16, Hep G2, BGC-823, MB-231 and A2780 cells were determined by MTT assay . Results: The results showed that the contents of Rubescens A, Erigeron, Erigeron and Ampelopsis B were respectively 0.085 6 ~ 0.856,0.014 04 ~ 0.140 4,0.070 8 ~ 0.708,0.018 98 ~ 0.189 8 g · L ~ (-1) showed a good linear relationship, the average recovery rates were 101.78%, 101.70%, 100.23%, 104.60% respectively. 4 diterpenes showed a certain degree of inhibition of cell proliferation, which Oridonin, furunll prime five kinds of cell proliferation inhibitory rates were> 50%. Rubescensine A, Mao Kui Li Su on the 5 tumor cell lines were better than the cytotoxicity of Mao Ye Cha Chae Su and Cha Cha B. The median inhibitory concentration (IC50) were 4.10 ~ 6.46, 2.34 ~ 4.77μmol·L -1, respectively. Conclusion: The method is simple, sensitive and reproducible, and can be used for the quality control of diterpene which inhibits the proliferation of major cells in Xiang tea.