Qualitative and Quantitative Analysis of Banhasasim-Tang Using UHPLC-Q-Orbitrap-MS and UHPLC-TQ-MS/MS

In this study, we analyzed the components of Banhasasim-tang (BHSST), a traditional prescription used to treat gastrointestinal disorders. Qualitative analysis was performed using ultra-high-performance liquid chromatography/quadrupole Orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS), and a total of 42 compounds were accurately and unambiguously identified by comparison with the corresponding reference standards and mass spectrometry (MS) results. A simultaneous quantitative method for analysis of the 22 identified compounds was established using UHPLC coupled with a triple-quadrupole tandem mass spectrometer (UHPLC-TQ-MS/MS) in multiple reaction monitoring mode. The established method was validated in terms of linearity (R2, 0.9990-0.9996), recovery (RSD, 0.37-3.60%), and intraday/interday precision (RSD, 0.35-8.90%). This method was applied to three batches of BHSST samples and revealed that two flavonoids from S. baicalensis, baicalin and wagonoside, were the most abundant components. This study presents the first comprehensive method for simultaneously identifying and quantifying the components of BHSST. Our method offers a valuable tool for the quality control of BHSST and serves as a foundational reference for further research on similar preparations.