基于UPLC-Q-TOF HRMS技术建立逍遥丸指纹图谱及化学成分鉴定

Fingerprint and identification of chemical constituents in Xiaoyao pills based on UPLC-Q-TOF HRMS

目的建立逍遥丸的UPLC-Q-TOF HRMS指纹图谱并对其主要化学成分进行分析鉴定。方法制备12批逍遥丸醇提物样品,采用Agilent ZORBAX Eclipse Plus C18(3.0 mm×100 mm,1.8μm)色谱柱,流动相为乙腈(A)-0.1%甲酸溶液(B)梯度洗脱,流速为0.4 mL·min^(-1),进样量为2μL。利用中药色谱指纹图谱相似度评价软件计算12批次逍遥丸样品指纹图谱相似度,评价不同批次样品间的质量差异,并对UPLC-Q-TOF HRMS技术采集的分子离子峰信息进行分析,通过化合物的精确相对分子质量、保留时间、质谱碎片信息以及对照品比对,分析逍遥丸中主要化学成分。结果建立12批逍遥丸LC-MS指纹图谱的共有模式,各共有峰间分离度好,12批逍遥丸指纹图谱相似度均在0.982以上,说明不同批次制备工艺较为稳定。采用UPLC-Q-TOF-MS技术鉴定了逍遥丸指纹图谱的35个共有峰的化学成分,并指认了11个成分,其中包括芍药内酯苷、芍药苷、绿原酸等。结论该方法的精密度、准确度、稳定性和重复性良好,建立的LC-MS指纹图谱为逍遥丸进一步质量控制和药效物质基础研究提供理论依据。

Objective To establish the fingerprints of Xiaoyao pills and identify its main chemical components based on UPLC-Q-TOF HRMS.Methods Twelve batches of alcohol extract samples of Xiaoyao pills were prepared.The separation was performed on Agilent ZORBAX Eclipse Plus C18(3.0 mm×100 mm,1.8μm),and the gradient elution of acetonitrile-methanol-0.1%formic acid was used as the mobile phase at a flow rate of 0.4 mL·min^(-1).The sample injection volume was 2μL.Similarity Evaluation System for Chromatographic Fingerprint of TCM was used to evaluate the similarity of 12 batches of sample fingerprints.The chemical components were analyzed by the liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF HRMS).The compounds were identified by relative molecular mass,retention time,mass spectrometry fragmention and comparison with standards.Results The similarity of 12 batches of Xiaoyao pills was over 0.982,indicating stable preparation process for different batches.Totally 35 compounds were identified by UPLC-Q-TOF-HRMS,the fingerprints of Xiaoyao pills were established with 11 common peaks,including albiflorin,paeoniflorin and chlorogenic acid,etc.Conclusion The established reference fingerprints of Xiaoyao pills are highly accurate and well separated,which can provide an important basis for the quality control and pharmacodynamic material basis study of Xiaoyao pills.