Objective This study aimed to systematically analyze the chemical constituents of the classical Chinese herbal formula Yinchen Wuling San (YCWLS) and identify its prototype components absorbed into the bloodstream using ultra-performance liquid chromatography coupled with Orbitrap tandem mass spectrometry (UPLC-Orbitrap-MS/MS). Methods The standard aqueous extract of YCWLS was prepared by decoction. UPLC-Orbitrap-MS/MS, combined with a self-built chemical database, was employed for qualitative analysis of the chemical constituents in the extract. A rat gavage administration experiment was conducted, and serum samples were collected. Prototype components absorbed into the bloodstream and metabolites were screened and identified by comparing the total ion chromatograms of blank serum and drug-containing serum. Results A total of 192 chemical constituents were identified in the aqueous extract of YCWLS, encompassing diverse structural types such as phenylpropanoids, flavonoids, coumarins, fatty acids, and nucleosides. Among these, 17 components were absorbed into the bloodstream in their prototype forms, including chlorogenic acid, 3-hydroxybenzyl alcohol, feruloyl putrescine, 2,5-dihydroxycinnamic acid, 2-methoxycinnamic acid, and 1-caffeoylquinic acid, etc. Conclusion This study comprehensively characterized the chemical composition group of the standard aqueous extract of YCWLS, elucidated its prototype components absorbed into the bloodstream and speculated its serum metabolites. The findings provide a crucial foundation for understanding the pharmacodynamic material basis and mechanism of action of this formula. Additionally, the methodology established herein offers a valuable reference for the modern research of other classical Chinese herbal prescription.

Yinchen Wuling San  /  blood-absorbed components  /  UPLC-MS/MS  /  chlorogenic acid  /  3-hydroxybenzyl alcohol  /  feruloyl putrescine  /  2,5-dihydroxycinnamic acid  /  2-methoxycinnamic acid  /  1-caffeoylquinic acid