UPLC-MS/MS法结合化学计量学分析万氏牛黄清心丸中14种胆汁酸成分  

Analysis of Fourteen Bile Acids in Wanshi Niuhuang Qingxin Pills by UPLC-MS/MS Combined with Chemometrics

作　　者：陈涛 林诗铃 林逸凡 陈雪婷 郑燕芳[1] 黄鸣清[1] CHEN Tao;LIN Shiling;LIN Yifan;CHEN Xueting;ZHENG Yanfang;HUANG Mingqing(Collegeof Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122 Fujian,China)

机构地区：[1]福建中医药大学药学院,福建福州350122

出　　处：《中药新药与临床药理》2023年第9期1271-1278,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(81973437,82274080);福厦泉国家自主创新示范区协同创新平台专项(2021FX02)。

摘　　要：目的建立超高效液相色谱-串联质谱(UPLC-MS/MS)法同时测定万氏牛黄清心丸中14种胆汁酸成分(牛磺胆酸、7-酮脱氧胆酸、甘氨胆酸、3-氧代脱氧胆酸、牛磺鹅去氧胆酸、牛磺脱氧胆酸钠、胆酸、甘氨鹅脱氧胆酸、甘氨脱氧胆酸、牛磺石胆酸钠、甘氨石胆酸、鹅去氧胆酸、去氧胆酸、石胆酸)的含量,并结合化学计量学考察不同批次万氏牛黄清心丸胆汁酸成分的差异。方法采用Hypersil GOLD色谱柱(2.1 mm×100 mm,1.9μm);流动相为甲醇-0.1%甲酸水,梯度洗脱;流速:0.2 mL·min^(-1);柱温:40℃;进样量:2μL。采用加热电喷雾离子源(HESI)负离子模式,平行反应监测(PRM)模式。以聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)评价不同批次万氏牛黄清心丸的胆汁酸成分差异。结果万氏牛黄清心丸中14种胆汁酸成分在相应测定范围内线性关系良好(r>0.9993),平均加样回收率为95.6%~103.6%,RSD为0.52%~4.62%。牛磺胆酸、甘氨胆酸、去氧胆酸、胆酸可以作为差异性成分对不同批次万氏牛黄清心丸进行区分。结论该方法能够实现对万氏牛黄清心丸中14种胆汁酸成分的快速定量分析,并可客观反映不同批次万氏牛黄清心丸之间的胆汁酸类成分含量差异。ObjectiveTo establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the simultaneous determination of 14 bile acids(taurocholate acid,7-ketodeoxycholic acid,glycocholic acid,3-oxocholic acid,taurochenodeoxycholic acid,sodium taurodeoxycholate,cholic acid,glycochenodeoxycholic acid,glycodeoxycholic acid,taurolithocholic acid,glycolithocholic acid,chenodeoxycholic acid,deoxycholic acid,lithocholic acid)in Wanshi Niuhuang Qingxin Pills,and to investigate the differences of bile acid components in different batches of Wanshi Niuhuang Qingxin Pills by UPLC-MS/MS combined with chemometrics.MethodsChromatographic conditions were as the follow:the chromatographic column was Thermo Fisher Scientific Bremen Hypersil GOLD(2.1 mm×100 mm,1.9μm),the mobile phase was methanol-0.1%formic acid in water with gradient elution,the flow rate was 0.2 mL·min-1and the column temperature was 40℃.The injection volume was 2μL.Analytes were determined by mass spectrometer with a heated electrospray ionization source(HESI)and parallel reaction monitoring(PRM)in negative ion mode.Hierarchical cluster analysis(HCA),principal component analysis(PCA),and orthogonal partial least squares discriminant analysis(OPLSDA)were used to evaluate the bile acid components of different batches of Wanshi Niuhuang Qingxin Pills.ResultsThe linear relationships of 14 bile acid components of Wanshi Niuhuang Qingxin Pills were good in the corresponding determination range(r>0.9993).The average recoveries varied from 95.6%-103.6%with the RSDs of 0.52%-4.62%.Taurocholate acid,glycocholic acid,deoxycholic acid,and cholic acid were the differential components to distinguish different batches of Wanshi Niuhuang Qingxin Pills.ConclusionThis method can achieve rapid qualitative and quantitative detection of 14 bile acid components in Wanshi Niuhuang Qingxin Pills,and can objectively reflect the difference of bile acid components in different batches of Wanshi Niuhuang Qingxin Pills.

关 键 词：万氏牛黄清心丸 超高效液相色谱-串联质谱 胆汁酸 化学计量学 

分 类 号：R284.1[医药卫生—中药学]