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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。
OBJECTIVE: To establish a method for the quantitative analysis of Corydalis Rhizoma by HPLC. Methods Human umbilical vein endothelial cells (HUVECs) were selected as target cells to extract the active ingredients of Corydalis. Corydalis freeze-dried powder was added to the logarithmic growth phase and incubated for 2 h. Cells were harvested and lysed with liquid nitrogen to obtain cell samples after drug extraction. It was detected by UPLC / Q-TOF-MS, and the substances extracted into the cells were analyzed, that is, the active ingredients entering into the cells of Corydalis to further exert their efficacy. Finally, the quantitative analysis of the active ingredients entering the cells was performed by HPLC. Results HUVEC extracted five pharmacodynamic components, namely tetrahydropalmatine, tetrahydroberberine, fizarmatine, berberine hydrochloride, hydrogen decontamination factor. At the same time the establishment of five components of the determination method. Conclusion The analysis of target cell extraction can reflect the active substance that exerts efficacy in the drug through the interaction between the compound and the cell, and plays an important role in the study of complex Chinese medicine.