Untargeted UHPLC-Q-Exactive Orbitrap MS and Targeted UHPLC-QqQ-MS/MS for Exploring the Effective Components of "Wen...

"Wen Tong Plaster" is renowned for its therapeutic effects of warming meridians, dispelling cold, and promoting blood circulation. However, its chemical composition and active pharmacological substances have yet to be fully elucidated. In this study, the chemical components of "Wen Tong Plaster" cataplasm and the transdermal components in vivo and in vitro were comprehensively analyzed by mass spectrometry targeted quantification method, and the release kinetics of the main transdermal components in vitro were evaluated, and the concentration-time variation of key active transdermal components in vivo was studied, finally molecular docking technology was used to predict the mechanism of action of key bioactive transdermal components. Using UHPLC-Q-Exactive Orbitrap MS, a total of 129 plant chemical components were found in the "Wen Tong Plaster" cataplasm extract, with 41 components detected as in vitro transdermal compounds. In subcutaneous tissue and uterine tissue, 53 and 36 in vivo transdermal components were characterized, respectively. The release kinetics of "Wen Tong Plaster" cataplasm were evaluated by tetrahydropalmatine, corydaline, and dihydrotanshinone I, three marker components, and fitted to a first-order kinetic model based on integrated release kinetics. After "Wen Tong Plaster" cataplasm administration, tetrahydropalmatine, corydaline, dihydrotanshinone I, and α-cyperone were accurately quantified in the subcutaneous tissue, plasma, and uterus tissue by UHPLC-QqQ-MS/MS, while the concentration-time changes of them in the subcutaneous tissue, plasma, and uterus tissue were investigated. The molecular docking results indicated that all four components exhibited favorable binding affinity with the targets, particularly demonstrating strong binding capacity to the cytochrome P450 1A1 target, suggesting it as a potential therapeutic target for "Wen Tong Plaster" cataplasm. In summary, the study revealed the pharmacodynamic material basis and in vitro release kinetics of "Wen Tong Plaster" cataplasm, providing a theoretical foundation and data support for further research on the mechanism of action and clinical application.