Application of UPLC-Q-TOF-MS with chemometric analysis and network pharmacology for comparison of different species: as an example

Paeonia lactiflora Pall. is a perennial herb of the buttercup family with a long history of medicinal use in China. With population migration and geographical changes, it gradually differentiates into different species, namely Paeoniae Radix Alba (PRA) and Paeoniae Radix Rubra (PRR). This study established an integrated strategy to investigate the different species of Paeonia lactiflora Pall. through ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), chemometric analysis combined with network pharmacology. Firstly, a total of 36 compounds were identified using UPLC-Q-TOF/MS, mainly including monoterpene glycosides, polyphenols, and phenolic acids. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) indicated that the studied samples were classified into PRA and PRR based on their species, while PRA or PRR could be classified as corresponding to geographical origins and morphological characteristics. Moreover, the orthogonal partial least squares discriminant analysis (OPLS-DA) and counter propagation artificial neural networks (CP-ANN) were applied to confirm classification results and obtain the chemical markers on different samples. Network pharmacology and molecular docking techniques demonstrated that active molecules have a higher affinity for ACTB and AKT1. The results of the content of representative components demonstrated differences between the two species. In conclusion, the methodology and results of this study provided a reliable basis for the identification and rational utilization of PRA and PRR, which were important for the appropriate clinical use and quality assurance of pharmaceutical materials. An integrated strategy was established to investigate the different species of Paeonia lactiflora Pall. through UPLC-Q-TOF/MS, chemometric analysis combined with network pharmacology.