活细胞固相色谱法联合高分辨质谱快速筛选龙血竭中镇痛活性成分

目的采用活细胞固相色谱法及高分辨液质联用技术快速筛选鉴定龙血竭中潜在的镇痛活性成分。方法首先采用HPLC-DAD-TOF-MS法对龙血竭药材中的主要化学成分信息进行表征,进一步选择以疼痛相关的离子通道受体表达丰富的小鼠背根神经元细胞为靶细胞,先使龙血竭提取液中的活性成分与靶细胞特异性结合,应用液质联用技术快速鉴定与细胞靶向亲和的化学成分。结果从龙血竭药材中鉴定出21个具有不同结构类型的主要成分,采用活细胞固相色谱法共筛选出10种能与背根神经元细胞亲和的潜在活性成分,包括二苯乙烯类2个、高异黄酮2个、高异黄烷1个、二氢查耳酮2个、黄酮寡聚体3个等。结论应用活细胞固相色谱法与高分辨质谱联用技术可快速筛选辨识龙血竭中活性成分,为龙血竭镇痛等活性成分的进一步研究奠定基础,也可为中药药效物质研究提供有益的方法学参考。

Rapid screening of potential analgesic ingredients from Draconis Resina by live cell immobilized chromatography coupled with HPLC-DAD-TOF-MS

Objective To rapidly screen the potential analgesic ingredients from DraconisResina by live cell immobilized chromatography coupled with HRMS. Methods An HPLC-DAD-ESI-TOF-MS technique was used to rapidly identify the main chemical constituents from DraconisResina. Based on the bio-specific affinity adsorption of bioactive compounds with receptors or channels on cells, the potential bioactive components in Draconis Resina could be selectively bound to the target cells-mice dorsal root neurons cells, then the chemical constituents with cell target affinity were identified by LC-HRMS. Results A total of 21 compounds with various structures were tentatively identified and characterized by HPLC-DAD-ESI-TOF-MS, and among them, 10 potentially analgesic active ingredients in Draconis Resina extract combined with dorsal root neurons cells were successfully detected and identified, including two stilbene, two homoisoflavones, one homoisoflavone, two dihydrochalcone, and three flavonoid oligomers. Conclusion Live cell immobilized chromatography coupled with LC-DAD-HRMS analysis could provide a rapid and efficient tool for finding the potential bioactive components in DraconisResina for the next pharmacology studies, which provide the reference for exploring of effective materials basis in Chinese medicines.