目的 基于超高效液相色谱-轨道阱高分辨质谱（UPLC-HR-Orbitrap-MS）和亲和超滤技术，筛选柴胡中的神经氨酸酶抑制剂，寻找具有神经氨酸酶抑制活性的单体并进行体外验证。方法 建立柴胡提取物的UPLC-HR-Orbitrap-MS分析方法，结合亲和超滤实验筛选柴胡中能够与神经氨酸酶特异性结合的活性单体成分，通过质谱解析及对照品验证确定成分，对筛选的活性单体进行体外神经氨酸酶抑制检测，并采用分子对接分析潜在的对接位点。结果 在柴胡中共筛选出能够与神经氨酸酶特异性结合的单体化合物10个，包括皂苷类、黄酮类和酚酸类，其中6''-O-柴胡皂苷A等的神经氨酸酶体外抑制活性为49.53%～78.07%，分子对接结果与体外神经氨酸酶结果存在差异。结论 亲和超滤-液质联用技术能够快速筛选中药中特异性活性成分，为中药活性标志物的发现提供新思路。

Objective Based on ultrahigh performance liquid chromatography Orbitrap high resolution mass spectrometry (UPLC-HR-Orbitrap-MS) and affinity ultrafiltration technology, the monomers with neuraminidase inhibitory activity in Bupleuri Radix were screened, and then to verify in vitro. Methods The UPLC-HR-Orbitrap-MS analysis method of the extract of Bupleuri Radix was established. The active monomers in it which can specifically bind to neuraminidase were screened by affinity ultrafiltration. The components were determined by mass spectrometry and control product verification and the neuraminidase inhibitions of the selected active monomers were detected in vitro, and the potential docking sites were analyzed by molecular docking. Results A total of ten monomers, including saponins, flavonoids and phenolic acids, were screened for specific binding to neuraminidase in Bupleuri Radix. Among them, the neuraminidase inhibitory activities of 6''-O- saikosaponin A and others ranged from 49.53% to 78.07% in vitro, and the molecular docking results were different from those of neuraminidase in vitro. Conclusion Affinity ultrafiltration-LC-MS can rapidly screen the specific active components in traditional Chinese medicine, which provides a new idea for the discovery of active markers of traditional Chinese medicine.

R284.1

国家自然科学基金面上项目（81774173）;山东省高等学校青创人才引育计划（活性天然产物发现与开发研究团队）;山东省中医药科技发展计划（2019-0019，2019-0021）