五倍子抑制流感病毒神经氨酸酶活性及活性部位的UPLC-Q-TOF-MS分析

Anti-influenza Virus Activity of Galla Chinensis Extracts and UPLC-Q-TOF-MS Analysis on Its Effective Part

目的:研究五倍子的流感病毒神经氨酸酶抑制活性,并采用超高效液相色谱串联四级杆飞行时间质谱仪(UPLC-Q-TOF-MS)对其活性部位化学成分进行分析。方法:采用神经氨酸酶抑制剂筛选试剂盒检测五倍子乙酸乙酯、乙醇、石油醚及水提取物对流感病毒神经氨酸酶的抑制作用。采用Acquity UPLC BEH C_(18)色谱柱,以0.1%甲酸水-甲醇溶液为流动相,梯度洗脱,电喷雾离子源(ESI)负离子模式,扫描抗神经氨酸酶活性较强的乙酸乙酯及乙醇部位并采集数据。结果:质量浓度为75 mg·L^(-1)时,五倍子4个提取物均显示了一定的神经氨酸酶抑制活性。五倍子的乙酸乙酯、乙醇提取物活性较强,抑制率分别为69.64%,78.24%。从2个提取物中分析鉴定出18个化合物。其中五没食子酸酰基葡萄糖、鞣花酸、没食子酸显示不同程度的神经氨酸酶抑制活性,半数抑制浓度(IC50)分别为46.15,143.42,249.51μmol·L^(-1)。结论:该研究表明五倍子具有较强的抑制神经氨酸酶活性,通过UPLC-Q-TOF-MS明确了五倍子提取物中18个主要成分,为五倍子抗病毒活性研究及其作用机制提供了依据。

Objective：The anti-influenza virus activity of Galla Chinensis extracts was evaluated by an influenza virus neuraminidase（NA） inhibition assay.The constituents in the active extract were identified using ultra-high performance liquid coupled with quadrupole time-of-flight mass spectrometry（UPLC-Q-TOF-MS） major constituents were used to further evaluate the NA inhibitory activity.Method：Using Neuraminidase Inhibitors Screen Kit to screen the four extracts of Galla Chinensis with anti-influenza virus activity.The chromatographic separation was performed on a C_（18） column with a gradient elution of methanol-aqueous formic acid 0.1%.The mass spectrometer equipped with electron spray ionization source was used as detector and the data was collected under the negative ion modes.Result：The results showed that four extracts of Galla Chinensis both presented NA inhibition activity.The ethyl acetate extract（EA） and the ethanol extract（E） presented strong.NA inhibitory activity with inhibition ratio of 69.64% and 78.24%,respectively.Eighteen constituents were characterized in these two active extracts by UPLC-Q-TOF-MS analysis,and three constituents（pentagalloyl glucose,ellagic acid,gallic acid） exhibited high activity against the influenza virus,IC_（50） values of 46.15,143.42,249.51 μmol·L^（-1） respectively.Conclusion：Galla Chinensis has significant influenza virus neuraminidase inhibitory activity.Eighteen phenolic acids constituents were identified by UPLC-Q-TOF-MS.The results indicate that Galla Chinensis contains phenolic acids constituents as inhibitors of NA and provide a basis for anti-Influenza Virus Activity of Galla Chinensis study on its mechanism.