Pinpointing antitumor constituents of Weifuchun capsules through UPLC-Q-TOF-MS/MS fingerprinting, spectrum-effect relationship analysis and human gastric adenocarcinoma cell bioassay validation.

Weifuchun capsule (WFC) is extensively used for the prevention and treatment of gastric precancerous lesions and as an adjuvant therapy following gastric cancer surgery, yet the specific bioactive components responsible for its antitumor effects remain poorly defined. In this study, we aimed to preliminarily screen the antitumor active constituents of WFC by combining ultra‑performance liquid chromatography‑quadrupole time‑of‑flight tandem mass spectrometry identification with spectrum-effect relationship analysis, while also evaluating batch-to-batch consistency via chromatographic fingerprinting. WFC samples were subjected to ethanol-water extraction followed by sequential liquid-liquid partitioning and viability assays on human gastric adenocarcinoma (AGS) cells revealed that the dichloromethane fraction exhibited the highest inhibition rate (95.84%). The chemical composition of this active fraction was analyzed by UPLC-Q-TOF-MS/MS, and fingerprints were established for ten batches of samples. Hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were applied to identify differential components among the batches. The relationship between antitumor efficacy and chemical constituents of WFC was investigated using Pearson correlation analysis, grey correlation analysis (GRA), and orthogonal partial least squares regression (OPLS). Eleven key compounds closely related to antitumor activity were identified, nine of which were validated by AGS cell assays. Three ent‑kaurane diterpenoids i.e., glaucocalyxin A, glaucocalyxin B, and oridonin exhibited potent anti‑proliferative activity with half‑maximal inhibitory concentration values ranging from 12.92 to 21.46 μM, whereas the positive control 5‑fluorouracil yielded an IC₅₀ of 73.03 μM under identical experimental conditions. Isorhamnetin, diosmetin, and eriodictyol exhibited moderate inhibitory effects, while naringin, neohesperidin, and hyperoside showed the weakest inhibition.