摘要
目的:明确甘青乌头抗乙酰胆碱酯酶的活性成分,为寻找新的抗阿尔茨海默病(AD)药物提供参考。方法:采用改良的Ellman’s法筛选甘青乌头抗乙酰胆碱酯酶活性部位,超高效液相色谱-四极杆飞行时间质谱法(UPLC-Q-TOF-MS/MS)对活性部位的化学成分进行分析,色谱条件为ACQUITY UPLC BEH C18色谱柱(2.1 mm×50 mm,1.7μm),流动相选择乙腈(A)-0.4%氨水溶液(B)梯度洗脱,流速0.4 mL·min^(-1),柱温30℃。二氯甲烷萃取部位A相比例随时间变化为0~3 min,5%A;3~7 min,5%~20%A;7~11.5 min,20%~33%A;11.5~15.5 min,33%~50%A;15.5~20.5 min,50%~80%A;20.5~23 min,80%~85%A;23~25 min,85%~95%A。正丁醇萃取部位A相比例随时间变化为0~2 min,5%A;2~8 min,5%~20%A;8~11 min,20%~33%A;11~15 min,33%~95%A。质谱条件为电喷雾离子源,正离子全扫描模式采集数据,检测范围m/z 100~1 500。结果:甘青乌头醇提物的二氯甲烷萃取部位和正丁醇萃取部位对乙酰胆碱酯酶均有一定的抑制作用,二者的半数抑制浓度分别为(64±4.4),(85.7±3.8)mg·L-1。经UPLC-Q-TOF-MS/MS分析,从二氯甲烷萃取部位共推测出21个生物碱类成分,正丁醇萃取部位共推测出11个生物碱类成分;2个部位均含有的关附壬素为首次在甘青乌头中发现。结论:二萜生物碱是甘青乌头抗乙酰胆碱酯酶活性的主要药效物质,值得深入挖掘。
Objective: To identify the anti-acetylcholinesterase active ingredients in Aconitum tanguticum,so as to lay the foundation for finding new anti-Alzheimer’s disease(AD)drugs. Method:The antiacetylcholinesterase active fractions of A. tanguticum were screened by the modified Ellman’s method,and the chemical composition of the active fraction was analyzed by ultra-performance liquid chromatographyquadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS). The chromatographic separation was performed on an ACQUITY UPLC BEH C18 column(2.1 mm×50 mm,1.7 μm)with acetonitrile(A)-0.4%ammonia aqueous solution(B)as mobile phase for gradient elution,and the column temperature was set at30 ℃ with the flow rate of 0.4 mL·min^(-1). Phase A of the dichloromethane fraction changed with time as follows:0-3 min,5%A;3-7 min,5%-20%A;7-11.5 min,20%-33%A;11.5-15.5 min,33%-50%A;15.5-20.5 min,50%-80%A;20.5-23 min,80%-85%A;23-25 min,85%-95%A. Phase A of the n-butanol fraction changed with time as follows:0-2 min,5%A;2-8 min,5%-20%A;8-11 min,20%-33%A;11-15 min,33%-95%A. Mass spectrometry was performed on electrospray ionization,data were collected in positive ion mode,and the detection range was m/z 100-1 500. Result: Both the dichloromethane and n-butanol fractions had a certain inhibitory effect on acetylcholinesterase,their half inhibitory concentration(IC50)values were(64±4.4)mg·L-1 and(85.7±3.8)mg·L-1,respectively. By UPLC-Q-TOF-MS/MS analysis,a total of 21 alkaloids were identified from the dichloromethane fraction,and 11 alkaloids were identified from n-butanol fraction. Guan-fu base Ⅰ,found in both fractions,was first discovered in A. tanguticum. Conclusion: Diterpene alkaloids are the main anti-acetylcholinesterase substances of A. tanguticum,which is worth further exploration.
作者
王月
刘晓谦
梁曜华
冯伟红
李春
王智民
WANG Yue;LIU Xiao-qian;LIANG Yao-hua;FENG Wei-hong;LI Chun;WANG Zhi-min(Guangdong Pharmaceutical University,Guangzhou 510006,China;National Engineering Laboratory for Quality Control Technology of Chinese Herbal Medicines,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第20期130-137,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
中央级公益性科研院所基本科研业务费专项(ZXKT17039)
北京市自然科学基金项目(7132152)。
