Babaodan (BBD) is a traditional Chinese medicine (TCM) prescribed for various inflammatory diseases, including viral hepatitis and acute genitourinary tract infection. Like other TCMs, BBD is a multi-component formula whose chemical composition and mode of action are largely unknown. The current study identified the bioactive ingredients of BBD using ultrahigh-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) followed by mass spectrometry molecular networking analysis. Subsequently, network pharmacology analysis was performed to predict the potential targets and pathways regulated by BBD. Eventually, a panel of compounds was selected and examined for their anti-inflammatory effects using lipopolysaccharide-stimulated RAW264.7 cells. Eighty-six compounds, including saponins, bile acids, and fatty acids, were identified. Tumor necrosis factor-alpha was identified as a key molecule. Pathways in cancer, inflammatory bowel disease, and hepatitis were predicted to be the major regulatory pathways. The results from bioassays validated ginsenoside Rb1, ginsenoside Rd, deoxycholic acid, chenodeoxycholic acid, and taurochenodeoxycholic acid as novel bioactive ingredients in BBD with anti-inflammatory effects. In conclusion, our study explains the anti-inflammatory efficacy of BBD from both chemical and biological aspects, which provides a scientific basis for the clinical application of BBD in inflammation-related diseases.

八宝丹 (BBD) 是一种中药 (TCM)，用于治疗各种炎症性疾病，包括病毒性肝炎和急性泌尿生殖道感染。与其他中药一样，BBD 是一种多成分配方，其化学成分和作用方式在很大程度上是未知的。目前的研究使用超高效液相色谱结合四极杆飞行时间质谱 (UPLC-QTOF-MS) 以及质谱分子网络分析来鉴定 BBD 的生物活性成分。随后，进行网络药理学分析以预测BBD调控的潜在靶点和通路。最终，选择了一组化合物，并使用脂多糖刺激的 RAW264.7 细胞检查了它们的抗炎作用。八十六种化合物，包括皂苷、胆汁酸、和脂肪酸，进行了鉴定。肿瘤坏死因子-α被确定为关键分子。癌症、炎症性肠病和肝炎的途径预计是主要的调节途径。生物测定结果证实人参皂苷 Rb1、人参皂苷 Rd、脱氧胆酸、鹅去氧胆酸和牛磺鹅去氧胆酸是 BBD 中具有抗炎作用的新型生物活性成分。总之，我们的研究从化学和生物学两个方面解释了BBD的抗炎功效，为BBD在炎症相关疾病中的临床应用提供了科学依据。生物测定结果证实人参皂苷 Rb1、人参皂苷 Rd、脱氧胆酸、鹅去氧胆酸和牛磺鹅去氧胆酸是 BBD 中具有抗炎作用的新型生物活性成分。总之，我们的研究从化学和生物学两个方面解释了BBD的抗炎功效，为BBD在炎症相关疾病中的临床应用提供了科学依据。生物测定结果证实人参皂苷 Rb1、人参皂苷 Rd、脱氧胆酸、鹅去氧胆酸和牛磺鹅去氧胆酸是 BBD 中具有抗炎作用的新型生物活性成分。总之，我们的研究从化学和生物学两个方面解释了BBD的抗炎功效，为BBD在炎症相关疾病中的临床应用提供了科学依据。