Simultaneous Determination of HuoXueJieGu Decoction Chemical Composition, Plasma Components, and Tissue Distribution by Ultra‐High‐Performance Liquid Chromatography‐Quadrupole‐Orbitrap Mass Spectrometry

HuoXueJieGu decoction is a compound traditional Chinese medicine comprising over 10 distinct herbal ingredients. Clinically utilized for managing rheumatoid arthritis and other rheumatic conditions, its documented efficacy includes promoting blood circulation, resolving stasis, reducing swelling, alleviating pain, and facilitating bone healing. However, the ambiguous material basis of HuoXueJieGu decoction significantly impedes the advancement of its quality control protocols and in‐depth exploration of its underlying pharmacological mechanisms. To address this gap, a comprehensive analytical method was developed for the systematic characterization of the HuoXueJieGu decoction aqueous extract. This method utilized a gradient elution system with ACN and an aqueous solution of 0.1% formic acid as the mobile phase. A total of 143 compounds were identified in the HuoXueJieGu decoction aqueous extract in vitro, which were categorized as follows: 31 organic acids, 29 flavonoids, 15 amino acids, 14 phenylpropanoids, 14 phenols, 12 alkaloids, 11 sugars and glycosides, 8 terpenoids, and 9 others. Furthermore, 31 constituents were detected in the plasma, indicating their systemic absorption. Tissue distribution analysis revealed the presence of 24, 19, 23, 40, and 38 prototype compounds in the heart, liver, spleen, lung, and kidney, respectively. Furthermore, this study represents the first report on the systematic and rapid characterization of the chemical profile of HuoXueJieGu decoction. It is also the first to interpret the functional roles of its constituents within the framework of the traditional “monarch‐minister‐assistant‐guide” theory. These findings provide a crucial scientific basis for elucidating the material basis of its efficacy and for advancing its quality control standards.