目的 采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱（UHPLC-Q-Exactive Orbitrap MS）技术结合药效学方法，对栀子大黄汤古方和现代优化方中不同药材比例配伍的化学成分进行分析，建立酒精性肝损伤模型评估其药效作用，并筛选潜在肝保护活性成分。方法 采用ACQUITY UPLC HSST3（100 mm×2.1 mm，1.8 μm）色谱柱以0.1%甲酸水-0.1%甲酸乙腈为流动相进行梯度洗脱，在正、负离子模式下采集质谱数据，根据文献和数据库资料，并结合化合物的保留时间、精确相对分子质量及二级碎片信息，鉴定栀子大黄汤中的化学成分。用酒精性肝损伤小鼠模型比较两方的药效水平，结合谱效关系研究筛选与肝保护作用相关的活性成分。结果 在栀子大黄汤中表征了80个共有化学成分，包括有机（酚）酸类22种、黄酮类18种、环烯醚萜类11种、蒽醌类8种、异黄酮类6种等。药效结果表明，古方比例给药的栀子大黄汤小鼠丙氨酸氨基转移酶（alanine amino-transferase，ALT）、天冬氨酸氨基转移酶（aspartate transferase，AST）、三酰甘油（triglyceride，TG）水平均明显低于现代优化方（P＜0.05）。结合谱效关系分析筛选出栀子苷酸、原儿茶酸、隐绿原酸等与酒精性肝损伤密切相关的药效物质。结论 明确了栀子大黄汤古方较现代优化方在酒精性肝损伤方面具有更优的肝保护作用，并阐释了其发挥抗酒精性肝损伤药效的潜在活性成分。

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.

R284.1

国家自然科学基金资助项目（82304687）；上海白玉兰人才计划浦江项目（23PJD113）