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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(2.1mm×100mm,2.5μm)液相色谱柱;流动相为乙腈(A)和0.5%甲酸水(B),梯度洗脱:0~5min,5%A;5~25min,5%~95%A;25~30min,95%A。柱温25℃,流速0.8mL/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/z100~1 500),其毛细管电压分别为3 500V和4 000V,二级质谱碰撞能量为10~40eV。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。
OBJECTIVE To rapidly identify and identify the chemical constituents of the Chinese traditional medicine Compound Babaidan by ultra-performance liquid chromatography-electrospray tandem four-stage flight time-of-flight mass spectrometry (UHPLC-Q-TOF / MS). Methods A total of 547 compounds were collected from the databases of Babao Dan using Agilent-Formula-Database Generator software. The total ion chromatogram of the extracts from Babao Dan was collected by UHPLC-Q-TOF / MS. The accurate mass-to-charge ratio of chemical components was analyzed and identified by automatic comparison. The secondary debris voltage of quadrupole time-of-flight mass spectrometry (Q-TOF / MS) The chemical composition further confirmed. The chromatographic separation was performed on a Waters XBridge BEH C18 (2.1 mm × 100 mm, 2.5 μm) liquid chromatography column with a mobile phase of acetonitrile (A) and 0.5% formic acid in water (B), gradient elution: 0-5 min, 5% A ~ 25min, 5% ~ 95% A; 25 ~ 30min, 95% A. The column temperature was 25 ℃, the flow rate was 0.8mL / min and the post-column split ratio was 2:1. Mass spectrometry was performed with four-stage tandem high-resolution time-of-flight mass spectrometry. Electrospray ionization was performed in both positive and negative ion mode (range, m / z100-1,500) with capillary voltages of 3 500 V and 4 000 V respectively. Energy is 10 ~ 40eV. Results In the positive and negative ion modes, a total of 78 chemical constituents from Babaidan were identified, including saponins, bile acids and amino acids. Conclusion The analytical method based on UHPLC-Q-TOF / MS was established to rapidly identify the chemical constituents in the Chinese traditional medicine Compound Babaidan. The chemical substance basis of Babaidan was clarified, which provided a basis for the quality control and pharmacological mechanism of Babaidan Research laid the foundation.