摘要
目的:建立超高效液相色谱-四极杆/静电场轨道阱质谱(UPLC-Q/Exactive MS)对牛黄清心丸(局方)中的胆汁酸成分进行定性定量分析,并对不同批次的牛黄清心丸进行质量评价。方法:采用Thermo HYPERSIL GOLD柱,以0.1%甲酸水溶液-甲醇为流动相进行梯度洗脱;质谱在负离子模式下分别进行全扫描/数据依赖二级扫描(Full MS/dd-MS2)和平行反应监测(PRM)。采用SPSS和SIMCA软件对含量测定结果进行化学计量学分析。结果:从牛黄清心丸中共鉴定出16个胆酸类成分,在定量分析中,16个胆酸在各自浓度范围内线性关系良好,其平均加样回收率为94.7%~104.8%。层次聚类分析(HCA)将13批样品分为2类,样品S1~S3、S6、S7为Ⅰ类,样品S4、S5、S8~S13为Ⅱ类,与主成分分析(PCA)结果一致;正交偏最小二乘法判别分析(OPLS-DA)显示胆酸、牛磺胆酸和甘氨胆酸的变量重要性投影值(VIP)均>1,是引起产品质量差异的关键成分。结论:所建方法能快速测定牛黄清心丸中的16个胆汁酸,并结合化学计量学筛选出3个影响产品质量的差异性成分,为牛黄清心丸的质量控制及评价提供科学依据。
Objective:To develop ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry(UPLC-Q/Exactive MS)method for the qualitative and quantitative analysis of bile acids in Niuhuang Qingxin Pills(Prescription of the Bureau),and to evaluate the quality of different batches of Niuhuang Qingxin Pills.Methods:Chromatographic separation was performed on a Thermo HYPERSIL GOLD column with mobile phase consisting of 0.1%formic acid solution-methanol using gradient elution.The mass spectrum monitoring was performed by full scan/data-dependent MS2(Full MS/dd-MS°)acquisition and parallel reaction monitoring(PRM)in negative ion mode,respectively.SPSS and SIMCA software were employed to perform the chemometrics analysis for the content determination results.Results:Sixteen bile acids were identified from Niuhuang Qingxin Pills.In the quantitative analysis,the calibration curves for 16 bile acids had good linearity in their concentration ranges.The average recoveries were in the range of 94.7%-104.8%.Hierarchical cluster analysis(HCA)classified 13 batches of Niuhuang Qingxin Pills into 2 categories,sample S1-S3,S6 and S7 in category I,sample S4,S5 and S8-S13 in category II,which was consistent with the results of principal component analysis(PCA).The results of orthogonal partial least squares discriminant analysis(OPLS-DA)showed that the variable importance in projection(VIP)values of cholic acid(CA),taurocholic acid(TCA)and glycocholic acid(GCA)were all greater than I,which were the key indicators causing the difference of product quality.Conclusion:The method developed in this study can quickly identify and quantify the 16 bile acids in Niuhuang Qingxin Pills,and three differential compounds affecting the product quality were screened out by chemometrics,which provides a scientific basis for the quality control and quality evaluation of Niuhuang Qingxin Pills.
作者
林逸凡
林诗铃
陈涛
林思荣
林晨
郑燕芳
黄鸣清
LIN Yifan;LIN Shiling;CHEN Tao;LIN Sirong;LIN Chen;ZHENG Yanfang;HUANG Mingqing(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350100,China;Fujian Institute for Food and Drug Quality Control,Fuzhou 350001,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2024年第8期4024-4032,共9页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金面上项目(No.81973437,No.82274080)
福厦泉国家自主创新示范区协同创新平台专项(No.2021FX02)。
关键词
牛黄清心丸
超高效液相色谱-四极杆/静电场轨道阱质谱
胆酸
含量测定
化学计量学
质量控制
Niuhuang Qingxin Pills
Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry(UPLC-Q/Exactive MS)
Bile acid
Content determination
Chemometrics
Quality control
