基于液质联用技术的芪参颗粒化学组成测定及一致性分析  

Chemical composition determination and consistency analysis of Qishen Granules based on liquid chromatography-mass spectrometry technology

作　　者：刘璐璐 李月婷[1,2] 苏汝彬 潘绮雪 魏祖英 屈碧琼 刘明妍 刘洁[2,3] 贾志鑫 肖红斌 LIU Lu-lu;LI Yue-ting;SU Ru-bin;PAN Qi-xue;WEI Zu-ying;QU Bi-qiong;LIU Ming-yan;LIU Jie;JIA Zhi-xin;XIAO Hong-bin(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China;Research Center for Chinese Medicine Analysis and Transformation,Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院,北京100029 [2]北京中医药大学中药分析与转化研究中心,北京100029 [3]北京中医药大学北京中医药研究院,北京100029

出　　处：《中草药》2022年第8期2312-2323,共12页Chinese Traditional and Herbal Drugs

基　　金：国家“重大新药创制”科技重大专项资助项目(2019ZX09201004-001);北京中医药大学新教师启动基金项目(2021-JYB-XJSJJ002)。

摘　　要：目的 建立高效液相色谱-三重串联四极杆质谱联用(HPLC-QQQ-MS/MS)测定芪参颗粒中38种主要成分含量的方法,明确其化学组成,并对制剂批次间一致性进行分析。方法 HPLC采用Zorbax Eclipse Plus C_(18)(50 mm×2.1 mm,1.8μm)色谱柱,0.1%甲酸水溶液-0.1%甲酸乙腈溶液梯度洗脱,质谱对正(11个离子对)、负(22个离子对)离子进行动态多反应监测(dynamic multi-response detection,dMRM),实现多批次芪参颗粒中38种主要成分的含量测定。通过主成分分析(principal component analysis,PCA)和正交偏最小二乘-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA),筛选影响批次间一致性的成分。结果 建立的HPLC-MS/MS方法在15min内完成了38个成分的同时定量分析,12批次芪参颗粒样品中38个化合物的总量为34.923~47.148 mg/g,其中臣药丹参中的丹酚酸B的量最高(16.718 mg/g),其次是金银花中的绿原酸(3.550 mg/g),君药黄芪中黄芪甲苷、毛蕊异黄酮苷葡萄糖苷的质量分数分别为0.136、0.753 mg/g,黑顺片中3种双酯型生物碱的质量分数均低于0.003 mg/g,其余化合物的质量分数在0.004~2.935 mg/g。PCA结合OPLS-DA筛选出了5个对批次一致性影响最大的成分,依次为黄芪甲苷、黄芪皂苷Ⅲ、隐丹参酮、丹参酮Ⅱ;和二氢丹参酮Ⅰ。结论 所建立的分析方法专属性强,灵敏度高,为芪参颗粒质量控制和一致性分析提供了科学方法和依据。Objective To establish a high performance liquid chromatography coupled to triple quadrupole mass spectrometry method(HPLC-QQQ-MS/MS)for the determination of 38 main components in Qishen Granules(芪参颗粒),clarify their chemical composition and analyze the consistency between batches.Methods The chromatographic separation was performed on a Zorbax Eclipse Plus C18 column(50 mm×2.1 mm,1.8μm)for gradient elution,the mobile phase was 0.1%formic acid water-0.1%formic acid acetonitrile.The dynamic multi-response detection(dMRM)scanning mode was used in the mass spectrometry for 11 ion pairs in positive ion mode and 22 ion pairs in negative ion mode to achieve the determination of 38 characteristic components in multiple batches of Qishen Granules.The principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)of each batch of Qishen Granules were carried out to obtain the components that affected the consistency between batches.Results Based on the established HPLC-MS/MS method,the simultaneous quantitative analysis of 38 components was completed within 15 min.The total amount of 38 compounds in 12 batches of Qishen Granules samples was 34.923—47.148 mg/g.Among them,the content of salvianolic acid B(16.718 mg/g)in the minister medicine Dansen(Salvia miltiorrhiza)was the highest,the second was the chlorogenic acid(3.550 mg/g)in Jinyinhua(Lonicerae Japonicae Flos).The contents of astragaloside IV and calycosin-7-glucoside in the monarch medicine Huangqi(Astragali Radix)were 0.136 and 0.753 mg/g.The contents of the three diester-alkaloids in Fuzi(Aconiti Lateralis Radix Praeparata)were all less than 0.003 mg/g,and the contents of the remaining compounds were between 0.004 and 2.935 mg/g.PCA combined with OPLS-DA screened out five components that had the greatest impact on batch consistency,followed by astragaloside IV,astragaloside III,cryptotanshinone,tanshinone II_(A) and dihydrotanshinone I.Conclusion The analytical method established in this study is specific and sensit

关 键 词：芪参颗粒 HPLC-QQQ-MS/MS 质量控制 主成分分析 正交偏最小二乘-判别分析 丹参素 哈巴苷 绿原酸 新绿原酸 隐绿原酸 咖啡酸 当药苷 芹糖甘草苷 甘草苷 异槲皮素 毛蕊花糖苷 异绿原酸C 迷迭香酸 芒柄花苷 异甘草苷 丹酚酸B 甘草素 丹酚酸A 毛蕊异黄酮 哈巴俄苷 异甘草素 黄芪皂苷Ⅲ 芒柄花素 甘草酸 二氢丹参酮Ⅰ 隐丹参酮 丹参酮Ⅰ 丹参酮Ⅱ 毛蕊异黄酮葡萄糖苷 木犀草苷 黄芪甲苷 苯甲酰新乌头原碱 苯甲酰乌头原碱 苯甲酰次乌头原碱 新乌头碱 乌头碱 次乌头碱 异毛蕊花糖苷 

分 类 号：R286.02[医药卫生—中药学]