Simultaneous quantification of 11 active constituents in Shexiang Baoxin Pill by ultraperformance convergence chromatography combined with tandem mass spectrometry

On account of the complexity of chemical constituents of Shexiang Baoxin Pill (SBP), a famous traditional Chinese medicine (TCM) formula, a novel and effective UPC2-MS/MS method was developed to simultaneously determine the content of 11 active compounds of SBP with outstanding separation ability. Eleven components in SBP, including 2 ginsenosides, 2 bile acids, 3 bufadienolides and 4 volatiles were detected by electrospray ionization tandem mass spectrometry in positive and negative ion modes with multiple reaction monitor (MRM). The analysis was performed at 30 °C using an Acquity UPC2 Diol (3.0 × 50 mm, 1.7 μm) column with linear gradient elution (eluent A, CO2; eluent B, methanol containing 20 mM ammonium acetate), back pressure of 2000 psi, flow rate of 1.2 mL/min and the injection volume of 1.0 μL. The method was extensively validated regarding the linearity (r ≥ 0.9974), precision (≤3.11%), recovery (93.34-104.50%), repeatability (≤2.00%) and stability (≤4.20%). Using this method, 11 active compounds of SBP with different polarity were simultaneously quantified in one chromatography analysis within 8 min. Statistical analysis of the effects of 11 compounds on the quality of SBP revealed that the content of cinnamaldehyde varied widely in different batches. This work presents an exemplary study for quality control of complex samples, especially for TCMs.