ABSTRACT In this study, we addressed the longstanding challenge of elucidating the complex material basis of traditional Chinese medicine (TCM) formulas by establishing an integrated, multi-platform UHPLC-QTOF MSE data-analysis workflow for Tongqiao Huoxue Decoction (THD), a prescription widely used in the clinical treatment of cerebral infarction. Unlike conventional single-software pipelines that often suffer from incomplete spectral libraries, limited algorithmic coverage, and platform-dependent biases, our strategy combined UNIFI, MS-DIAL, QI, and FBMN to jointly process the data, thereby reducing the analytical blind spots and interference inherent to any single tool. This multi-angle identification system enabled a high-confidence characterization of 235 chemical constituents in THD and 101 in vivo metabolites detected in serum and urine samples of rats, while also revealing predominant metabolite classes potentially associated with THD's pharmacodynamic mechanisms in cerebral infarction. The resulting workflow demonstrated high accuracy, reproducibility, and efficiency, offering a rigorous and comprehensive solution for biological and medicinal sample analysis. Overall, this work not only deepens the understanding of THD's therapeutic material foundation, but also provides a novel, generalizable methodological paradigm that advances mass-spectrometry-based characterization of multi-component TCM prescriptions and their in vivo transformation profiles.

SYDNEY--Mr. Arthur Fletcher, of Harcourt Parade, Rosebery, thought he say a "flying saucer" on his way to work