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| 中药复方制剂八宝丹中化学成分的UHPLC-Q-TOF/MS分析 |
| 李洋1,余静1,赵亚红2,安泳潼2,薛丹2,柴逸峰3,张国庆1,张海1* |
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(1. 第二军医大学东方肝胆外科医院药材科, 上海 200438;
2. 上海医药集团股份有限公司中央研究院, 上海 200123;
3. 第二军医大学药学院药物分析学教研室, 上海 200433
*通信作者) |
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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。 |
| 关键词: 超高效液相色谱法 四级杆飞行时间串联质谱法 八宝丹 中药分析化学 中药理化鉴定 |
| DOI：10.16781/j.0258-879x.2016.12.1548 |
| 投稿时间：2016-06-22修订日期：2016-09-23 |
| 基金项目:上海市科委产学研医结合项目（15DZ1900102）. |
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| UHPLC-Q-TOF-MS in analyzing chemical constituents of traditional Chinese herbal preparation Babao Dan |
| LI Yang1,YU Jing1,ZHAO Ya-Hong2,AN Yong-tong2,XUE Dan2,CHAI Yi-feng3,ZHANG Guo-qing1,ZHANG Hai1* |
(1. Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China;
2. Central Research Institute, Shanghai Pharmaceutical(Group) Co., Ltd., Shanghai 200123, China;
3. Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
* Corresponding author) |
| 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. |
| Key words: ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry Babao Dan analytical chemistry (TCD) TCD physic chem identific |
