利用分子量印迹快速筛选活心丸中蟾蜍二烯酸内酯和二萜生物碱类化合物

Rapid Screening of Bufadienolides and Diterpenoid Alkaloids in Huoxin Pills by Molecular Weight Imprinting

目的:建立分子量印迹(MWI)质谱数据后处理策略,快速筛选超高效液相色谱-四极杆-飞行时间质谱法(UPLC-Q-TOF-MS)采集的活心丸中蟾蜍二烯酸内酯和二萜生物碱类化学成分。方法:基于文献和PubMed等数据库,分别构建源于蟾酥和附子的蟾蜍二烯酸内酯和二萜生物碱类化合物内部数据库,将m/z[M+H]+按整数部分和小数部分拆分,以±0.005为误差范围,建立已知化合物二维散点图。采用UPLC-Q-TOF-MS获取m/z 50~1000的MS1和MS2数据,将采集到的MS1质荷比分成整数部分和小数部分,绘制样品二维散点图,与已知化合物二维散点图匹配,筛选已知化合物。并结合预测分子式及MS2信息,推导筛选化合物的质谱裂解途径,鉴定筛选出的化合物的化学结构。结果:从活心丸中初步筛选并鉴定了蟾酥来源的47个蟾蜍二烯酸内酯类化合物及附子来源的33个二萜生物碱类化合物。结论:利用MWI的数据后处理策略实现了对活心丸中蟾蜍二烯酸内酯和二萜生物碱类化合物的快速鉴定,为活心丸的质量控制及临床用药安全提供参考,也为解析中药化学成分的质谱数据提供了方法。

Objective:To rapidly screen out bufadienolides and diterpenoid alkaloids in Huoxin Pills(HXPs)from massive mass spectrometric data by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)based on the post-acquisition data processing strategy of molecular weight imprinting(MWI).Methods:Firstly,the in-house data library of bufadienolides from Bufonis Venenum and diterpenoid alkaloids from Aconiti Lateralis Radix Praeparata was built based on literatures and databases such as PubMed.The m/z[M+H]+were then divided into an integral part and a decimal part,resulting in a two-dimensional scatter plot of the known compounds with±0.005 as the error range.Subsequently,UPLC-Q-TOF-MS was utilized to obtain MS1 and MS2 spectral data with m/z 50-1000.The collected m/z of MS1 were also divided into an integer part and a decimal part to construct the two-dimensional scatter plot.After matching those two-dimensional scatter diagrams,the known compounds were rapidly screened out.Meanwhile,combined with the predicted molecular formula and MS2 spectral data,the mass fragmentation pathways of the screened compounds were proposed,and the chemical structures of the screened bufadienolides and diterpenoid alkaloids were confirmed.Results:Forty-seven bufadienolides from Bufonis Venenum and thirty-three diterpenoid alkaloids from Aconiti Lateralis Radix Praeparata were preliminarily identified in HXPs.Conclusion:This study has realized the rapid identification of bufadienolides and diterpenoid alkaloids in HXPs,which provides references for the quality control and clinical safety of HXPs and a method for analyzing mass spectrometric data of chemical components of Chinese medicine.