Strategy for identification of characteristic components from multiphase of Sanwu Huangqin Decoction via UHPLC-Q-TOF-MS and UHPLC-QQQ-MS combined with multivariate statistical analysis

Sanwu Huangqin Decoction (SHD), a classic prescription for nourishing yin and clearing heat, is commonly used to treat febrile diseases, erythromelalgia, autoimmune liver diseases, and skin diseases. However, research on the quality standard of SHD is still in its infancy and has focused mainly on single Chinese herbs, with few systematic studies. Currently, traditional Chinese medicine decoctions, as a complex polydisperse system, have attracted widespread attention. It is necessary to establish an integrated and rapid identification strategy for evaluating and controlling the quality of multiphase of SHD. The ingredients in the three phases of SHD were first isolated via the centrifugation-dialysis technique, and an method was established via ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) for discrimination. The identification results and multivariate statistical analysis revealed that the SHD and three phases were clearly clustered, whereas the composition difference between the nanophase state and SHD was the smallest. Moreover, 34 significantly common differential markers were screened out by variable importance for projection (VIP) > 1 and P < 0.05 to construct the orthogonal partial least squares-discriminant analysis (OPLS-DA) regression prediction model. Furthermore, a strategy for potential effective phase identification and quality evaluation index was established based on the absolute quantitative analysis of the 13 characteristic ingredients via ultrahigh-performance liquid chromatography triple quadrupole mass spectrometry (UHPLC-QQQ-MS). The comprehensive strategy of qualitative analysis combined with absolute quantitative analysis may be an effective tool for the identification and discrimination of SHD and the potential effective phase state. These findings could provide a significative reference for further revealing the material basis and determining the quality control indicators of SHD.