【摘要】 目的 建立高速逆流色谱法分离纯化红豆杉中10-DABⅢ类和紫杉醇类部位化合物的方法,分析化合物相关抗炎活性。方法 通过调整分离系统中的流速和溶剂比例筛选出最佳的高速逆流分离系统。薄层色谱法和高效液相色谱法鉴定和检测10-DABⅢ和紫杉醇含量。通过CCK-8法测定化合物用药的安全浓度。并用化合物作用于脂多糖诱导的巨噬细胞RAW264.7炎症模型,ELISA检测IL-10的分泌量。结果 高速逆流色谱最佳分离系统为正己烷-乙酸乙酯-甲醇-乙醇-水=5:5:5:1:6.5,薄层色谱和高效液相色谱法测定化合物1为含量41.32%的10-DABⅢ类部位化合物,化合物2为48.70%的紫杉醇类部位化合物。化合物1和化合物2在浓度3.125~12.5μg/ml间无明显细胞毒性。ELISA结果显示浓度为12.5μg/ml的10-DABⅢ类部位化合物促进IL-10分泌,10-DABⅢ类部位为抗炎活性成分部位。结论 此方法稳定、高效、经济实用,为红豆杉有效部位的提取分离提供有效的研究依据。更多还原

【Abstract】 Objective To develop a high-speed countercurrent chromatography method for the separation and purify the 10-DABIII and paclitaxel compounds in Taxus chinensis related to the anti-inflammatory activity of compounds.Methods Separation and purification were carried out by highspeed countercurrent,and the optimal high-speed countercurrent separation system was screened by adjusting the flow rate and solvent ratio in the separation system to purify 10-DABIII and paclitaxel-based compounds.Thin layer chromatography and high performance liquid chromatography were used to identify and detect 10-DABIII and paclitaxel content.The safe concentration of the two compounds was determined by the CCK-8 method.Different doses of compounds were applied to lipopolysaccharide-induced macrophage RAW264.7 inflammation model,and the secretion of IL-10 in cell supernatant was detected by ELISA.Results The best separation and separation system for high-speed countercurrent chromatography is n-hexane-ethyl acetate-methanolethanol-water=5:5 : 5:1:6.5.The Thin layer chromatography and high performance liquid chromatography were used to determine the compound of the 10-DABIII type compound having a compound 1 content of 41.32%,and the compound 2 was a 48.70% paclitaxel site compound.Compound 1 and Compound 2 showed no significant cytotoxicity to cells at concentrations between 3.125 and 12.5 μg/ml.The 10-DABIII class compound significantly promoted IL-10 secretion at a concentration of 12.5 μg/ml,10-DABIII site was an anti-inflammatory active ingredient.Conclusion The method is stable,efficient,economical and practical,and provides an effective research basis for the extraction and separation of effective parts of Taxus.更多还原