UHPLC-Q-TOF-MS in analyzing chemical constituents of traditional Chinese herbal preparation Babao Dan / 第二军医大学学报

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 flightmass 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 Baboo 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 Baboo 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 X 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°C; the flowing rate was 0.8 mL/min; and the post-column spit ratio was 21. 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 Dnn, including saponins, bile acids, amino acids and so on. Conclusion A rapid and efficient method for identifying the chemicaf 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.