Chemical identification and metabolic profiling of Tongmai granules using UHPLC-QTOF-MS-based molecular networking and modified mass defect filtering techniques

Abstract

Ischemic stroke (IS) is a major cause of death and disability worldwide, and its complicated biological processes make developing effective treatments challenging. Tongmai granules, the famous Chinese herbal formula, was good at treating IS. However, the active compounds and their underlying mechanisms are still not well understood. To elucidate the active compounds of Tongmai granules against IS, the chemical compounds of Tongmai granules were characterized using ultra-high performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS). Using molecular networking (MN) method, a total of 89 compounds were quickly identified. Afterwards, the metabolites profiling of Tongmai granules in the middle cerebral artery occlusion/reperfusion (MCAO/R) rats' plasma and brain were conducted using UHPLC-HRMS and mass defect filtering (MDF) strategy. And 80 metabolites (25 prototypes and 55 metabolites) were characterized in plasma and brain samples. Network pharmacology and molecular docking have identified 8 active components for treating IS, including daidzein, tanshinone IIA, puerarin, cryptotanshinone, tanshinone IIb, butylidenephthalide, senkyunolide A and salvianolic acid A. These components may exert their effects through regulating TP53, SRC, and STAT3. The study provides a comprehensive characterization of Tongmai granules and insights into their therapeutic mechanisms.