Objective Ultra-high-performance liquid chromatography coupled with quadrupole-electrostatic field orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap MS) technology combined with pharmacological methods was developed to analyze the chemical components of Zhizi Dahuang Decoction in different proportions in ancient and modern optimized prescriptions. Subsequently, an alcoholic liver injury model was established to evaluate its hepatoprotective effects and screen potential active ingredients in traditional Chinese medicine. Methods Gradient elution was performed on an ACQUITY UPLC HSS T3 (2.1 mm × 100 mm, 1.8 μm) column with 0.1% formic acid aqueous solution and acetonitrile. The mass spectrometry data were collected in positive and negative ion mode. Chemical constituents of Zhizi Dahuang Decoction were identified by comparing with literature and database records, considering retention time, accurate molecular mass, and MS/MS fragmentation patterns. Furthermore, an alcoholic liver injury mouse model was utilized to compare the efficacy of the two formulations, and to screen for active components related to hepatoprotective effects through a spectrum-activity relationship study. Results A total of 80 common chemical components were identified in Zhizi Dahuang Decoction, including 22 organic (phenolic) acids, 18 flavonoids, 11 iridoids, eight anthraquinones, six isoflavones, etc. Pharmacological results demonstrated that the classical formula group exhibited significantly lower serum levels of alanine amino-transferase (ALT), aspartate transferase (AST), and triglyceride (TG) compared to the modern optimized group (P < 0.05). Screening potential pharmacological substances closely related to alcohol-induced liver injury, such as geniposide acid, protocatechuic acid, and cryptochlorogenic acid, through spectrum-effect relationship analysis. Conclusion This study demonstrated that the ancient formulation of Zhizi Dahuang Decoction has better hepatoprotective effects on alcoholic liver injury than the modern optimized formulation, and further characterized the potential bioactive components contributing to its anti-alcoholic liver injury efficacy.

Zhizi Dahuang Decoction  /  alcoholic liver injury  /  active ingredients  /  geniposide acid  /  protocatechuic acid