[关键词]
[摘要]
目的 建立靶细胞萃取延胡索Corydalis Rhizoma中的药效成分并采用HPLC法进行定量分析的方法。方法 选用人脐静脉内皮细胞(HUVEC)作为靶细胞,萃取延胡索中的药效成分。在细胞对数生长期加入延胡索冻干粉,孵育2 h收集细胞并用液氮裂解,得到萃取药物后的细胞样品。采用UPLC/Q-TOF-MS对其进行检测,分析萃取进入细胞内的物质,即延胡索中进入细胞进一步发挥药效的有效成分。最后,用HPLC法对进入细胞中的有效成分进行定量分析。结果 HUVEC萃取到5个药效成分,分别为延胡索乙素、四氢小檗碱、延胡索甲素、盐酸小檗碱、去氢延胡索甲素。同时建立5个成分的含量测定方法。结论 靶细胞萃取的分析方法可以通过化合物与细胞之间的相互作用,反映出药物中发挥药效的活性物质,在中药复杂体系的研究中起到重要的作用。
[Key word]
[Abstract]
Objective To establish a method that using target cell to extract the pharmacodynamic components and quantitative analysis in Corydalis Rhizoma. Methods Take the human umbilical vein endothelial cells (HUVEC) as target cells, and add the freeze-dried powder of yanhusuo in logarithmic growth period. After 2 h incubation, the cells were collected and liquid nitrogen was used to cleave those cell samples. While compounds in the extract of cell samples were determined by UPLC/Q-TOF-MS, the effect of effective components that entered the cell of Corydalis Rhizoma was ananlyzed. Finally, HPLC was used to quantitatively analyze the effective components. Results After HUVEC extraction, five active ingredients were obtained, such as tetrahydropalmatine, berberine, tetrahydroberberine, berberine hydrochloride, and dehydrocorydalinecorydaline, respectively. At the same time, the method was established for the determination of the five components. Conclusion The analysis method of the target cell extraction can reflect the active substance in the medicine, through the interaction between the compound and the cell. This method will play an important role in the study of complex system of traditional Chinese medicine.
[中图分类号]
[基金项目]
国家科技支撑计划课题(2011BAI07B00)