中药复方制剂八宝丹中化学成分的UHPLC-Q-TOF/MS分析
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第二军医大学东方肝胆外科医院药材科,第二军医大学东方肝胆外科医院药材科,上海医药集团股份有限公司中央研究院,上海医药集团股份有限公司中央研究院,上海医药集团股份有限公司中央研究院,第二军医大学药学院药物分析教研室;第二军医大学药学院药物分析教研室,第二军医大学东方肝胆外科医院药材科,第二军医大学东方肝胆外科医院药材科;上海医药集团股份有限公司中央研究院;第二军医大学药学院药物分析教研室;第二军医大学药学院药物分析教研室

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上海市科委产学研医结合项目(15DZ1900102).


UHPLC-Q-TOF-MS in analyzing chemical constituents of traditional Chinese herbal preparation Babao Dan
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Department of Pharmacy,Eastern Hepatobiliary Surgery Hospital,Second Military Medical University,Shanghai,Department of Pharmacy,Eastern Hepatobiliary Surgery Hospital,Second Military Medical University,Shanghai,Central Research Institute of Shanghai Pharmaceutical Group Co,Ltd,Central Research Institute of Shanghai Pharmaceutical Group Co,Ltd,Central Research Institute of Shanghai Pharmaceutical Group Co,Ltd,School of Pharmacy,Second Military Medical University,Shanghai,Department of Pharmacy,Eastern Hepatobiliary Surgery Hospital,Second Military Medical University,Shanghai,Department of Pharmacy,Eastern Hepatobiliary Surgery Hospital,Second Military Medical University,Shanghai,;;Central Research Institute of Shanghai Pharmaceutical Group Co,Ltd;School of Pharmacy,Second Military Medical University,Shanghai

Fund Project:

Supported by Combination Project of Industry, University, Research and Medicine of Shanghai Science and Technology Commission (15DZ1900102).

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    摘要:

    目的 采用超高效液相色谱-电喷雾串联四级杆飞行时间质谱(UHPLC-Q-TOF/MS)分析技术对中药复方制剂八宝丹中的化学成分进行快速分析鉴别。方法 采用安捷伦“Formula-Database Generator”软件自建包括547个化合物的八宝丹化学成分数据库,运用UHPLC-Q-TOF/MS技术采集八宝丹提取物的总离子流色谱图,根据各化学成分的精确质荷比,通过自动比对的方法进行分析鉴别;进一步调节四级杆飞行时间质谱(Q-TOF/MS)的二级碎片电压,根据二级裂解碎片离子对八宝丹中的化学成分进一步确证。色谱分离采用Waters XBridge BEH C18(2.1 mm×100 mm,2.5 μm)液相色谱柱;流动相为乙腈(A)和0.5%甲酸水(B),梯度洗脱:0~5 min,5% A;5~25 min,5%~95% A;25~30 min,95% A。柱温25℃,流速0.8 mL/min,柱后分流比为2:1。质谱分析采用四级杆串联高分辨飞行时间质谱,电喷雾离子源正、负离子模式下扫描(范围均为m/z 100~1 500),其毛细管电压分别为3 500 V和4 000 V,二级质谱碰撞能量为10~40 eV。结果 在正、负离子模式下,共鉴别出八宝丹中的化学成分78个,包括皂苷、胆汁酸、氨基酸等。结论 建立了基于UHPLC-Q-TOF/MS的分析方法,可对中药复方制剂八宝丹中的化学成分快速鉴别,阐明了八宝丹的化学物质基础,为八宝丹的质量控制及药理机制研究奠定了基础。

    Abstract:

    Objective To rapidly identify the chemical components in a traditional Chinese compound herbal preparation Babao Dan by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF/MS). Methods A formula database including 547 compounds was developed by Agilent software "Formula-Database Generator". The total ion chromatograms of the Babao Dan extracts were obtained by UHPLC-Q-TOF/MS, and the chemical components were identified by automatic matching method according to the exact mass-to-charge ratio of each chemical component. The chemical components of Babao Dan was further verified using the secondary fragment ion obtained by adjusting the secondary monitor voltage of Q-TOF/MS. The separation was performed on a Waters Xbridge BEH C18 column (2.1 mm×100 mm, 2.5 μm). The mobile phase composing of acetonitrile (A) and 0.5% formic acid aqueous solution (B) was used for gradient eluting (0-5 min, 5% A; 5-25 min, 5%-95% A; 25-30 min, 95% A). The temperature of column was 25℃; the flowing rate was 0.8 mL/min; and the post-column split ratio was 2:1. The Q-TOF/MS and electro spray ion source (ESI) were applied for qualitative analysis under positive and negative ion mode, with the mass scan range being m/z 100-1 500. The capillary voltage was 3 500 V under positive ion mode and 4 000 V under negative ion mode, and the range of collision energy of MS2 was 10-40 eV. Results UHPLC-Q-TOF-MS identified a total of 78 chemical components from Babao Dan, including saponins, bile acids, amino acids and so on. Conclusion A rapid and efficient method for identifying the chemical components of Babao Dan by UHPLC-Q-TOF-MS has been established, and the chemical components of Babao Dan has been clarified, which lays a foundation for the quality control and further pharmacological study of Babao Dan.

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  • 收稿日期:2016-06-22
  • 最后修改日期:2016-09-23
  • 录用日期:2016-11-01
  • 在线发布日期: 2016-12-23
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