Unveiling the chemical components variation of Sishen formula induced by different prescription ratios by the advanced liquid chromatography/mass spectrometry approaches

Compatibility with the varying prescription ratios is a unique feature in the Chinese medicine practice. Sishen formula (SSF; containing Psoraleae Fructus-PF, Myristicae Semen-MyS, Schisandrae Chinensis Fructus-SCF, and Euodiae Fructus-EF) with different proportions has been used to treat the irritable bowel syndrome, however, the underlying chemical difference needs to be clarified, particularly the influence on extracting those known toxic components. To tackle this issue, an integrated strategy was developed: 1) systematic multicomponent characterization by an off-line two-dimensional liquid chromatography/ion mobility time-of-flight mass spectrometry (2D-LC/IM-QTOF-MS) approach with the HDMSE-HDDDA hybrid scan; 2) discovery of potential markers associated with SSF proportion variation by untargeted metabolomics; and 3) quantitative assays of representative markers by multiple reaction monitoring (MRM) to unveil the effects on the dissolution of toxic components. Consequently, orthogonal separation, high sensitivity of detection, and intelligent data interpretation, enabled the characterization of 233 compounds from the SSF decoction. Totally 50 differential components were discovered from SSF with 16 different ratios by high-definition LC-MS profiling and multivariate statistical analysis. An MRM approach was validated offering the absolute content alternation of 18 compounds in SSF. Increasing of SCF, MyS, and EF moderately could reduce the dissolution of the main toxic coumarins of PF, while increasing of SCF promoted dissolution of the main toxic alkaloids of EF, which could be restrained by increasing MyS. Conclusively, the in-depth multicomponent characterization of SSF was achieved, and the disparities in the compatibility of various SSF proportions were exposed, which may bridge to the functions/toxicity variations of SSF in clinic.