Xiaoer-Feire-Kechuan (XFK) is an 11-herb Chinese medicine formula to treat cough and pulmonary inflammation. The complicated composition rendered its chemical analysis and effective-component elucidation. In this study, we combined quantitative analysis and bioactivity test to reveal the anti-inflammatory constituents of XFK. First, UPLC-DAD and UHPLC/Q-Orbitrap-MS methods were established and validated to quantify 35 analytes (covering 9 out of 11 herbs) in different XFK formulations. Parallel reaction monitoring mode built in Q-Orbitrap-MS was used to improve the sensitivity and selectivity. Then, anti-inflammatory activities of the 35 analytes were analyzed using in vitro COX-2 inhibition assay. Finally, major analytes forsythosides H, I, A (8–10), and baicalin (15) (total contents varied from 21.79 to 91.20 mg/dose in different formulations) with significant activities (inhibitory rate ≥ 80%) were proposed as the anti-inflammatory constituents of XFK. The present study provided an effective strategy to discover effective constituents of multi-herb formulas.

小二肺热咳喘（XFK）是一种治疗咳嗽和肺部炎症的11种中药配方。复杂的成分使其化学分析和有效成分阐明。在这项研究中，我们结合定量分析和生物活性测试来揭示 XFK 的抗炎成分。首先，建立并验证了 UPLC-DAD 和 UHPLC/Q-Orbitrap-MS 方法以量化不同 XFK 制剂中的 35 种分析物（涵盖 11 种草药中的 9 种）。采用 Q-Orbitrap-MS 内置的平行反应监测模式来提高灵敏度和选择性。然后，使用体外 COX-2 抑制试验分析 35 种分析物的抗炎活性。最后，主要分析物连翘苷 H、I、A ( 8 – 10 ) 和黄芩苷 ( 15)（总含量从 21.79 到 91.20 毫克/剂在不同配方中变化）具有显着活性（抑制率 ≥ 80%）被提议作为 XFK 的抗炎成分。本研究提供了一种有效的策略来发现多草药配方的有效成分。