This study aimed to characterize the chemical profile of Danggui Buxue Decoction (DBD) and investigate the systemic exposure and tissue distribution of its constituents in mice following oral administration, using UHPLC-Q-Exactive Orbitrap high-resolution mass spectrometry (HRMS). Methods A total of 12 healthy male C57BL/6J mice were randomly divided into a blank control group (n = 3), administered purified water, and a DBD-treated group (n = 9), given a DBD extract at 16 g/kg by gavage. Serum and tissue samples were collected at 0.5, 1.5, and 3 h post-dose, pooled, and processed via protein precipitation. Chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column (100 mm × 2.1 mm, 1.7 μm) using a gradient elution with 0.1% formic acid in water and methanol as the mobile phase. Mass spectrometric data were acquired in both positive and negative ion modes via full-scan MS¹/data-dependent MS² scanning. Compound identification was achieved by analyzing relative retention times and MS¹ fragment information using Xcalibur 4.2 software, with confirmation against reference standards, literature data, and databases. Results A total of 158 chemical compounds were identified in DBD, comprising 56 flavonoids, 44 saponins, 24 phthalides, 17 organic acids, 8 coumarins, 2 alkaloids, and 7 other constituents. The number of prototype compounds detected in mice after DBD administration varied over time. The abundance of these migrating components across tissues followed the order: ileum > liver > serum > lung > kidney > spleen > heart > brain. Key constituents identified in the serum and various tissues included 7,3'-dihydroxy-5'-methoxyisoflavone (pterocarpan E), calycosin-7-O-β-D-glucoside, methylnissolin, calycosin, oroxylin A, isomucronulatol, 3-butylidenephthalide, (6αR,11αR)-3,9-dimethoxy-10-hydroxypterocarpan and formononetin. Conclusion This study delineates the comprehensive chemical composition of DBD and in vivo absorption and tissue distribution patterns using HRMS, providing crucial insights for identifying the bioactive constituents of DBD and supporting further tissue pharmacokinetic investigations.

UHPLC-Q-Exactive Orbitrap HRMS  /  famous classical formulas  /  Danggui Buxue Decoction  /  constituents absorbed into serum  /  tissue distribution  /  7,3'-dihydroxy-5'-methoxyisoflavone  /  calycosin-7-O-β-D-glucoside  /  oroxylin A  /  3-butylidenephthalide