IntroductionScreening of novel pancreatic lipase inhibitors from complex natural products is a meaningful task.ObjectivesThrough accurately screening and separating pancreatic lipase inhibitors from Clematis tangutica (C. tangutica), to discover new leading compounds for slimming and accelerate the development and utilization of Tibetan medicine resources.MethodsAn integrated strategy that combines affinity ultrafiltration and high‐performance liquid chromatography‐quadrupole time‐of‐flight mass spectrometry (AU‐HPLC‐QTOFMS), targeted separation, in vitro validation, and molecular docking was developed to screen pancreatic lipase inhibitors from C. tangutica. The AU‐HPLC‐QTOFMS technique was performed to fish for the potential active substances. Macroporous resin, preparative liquid chromatography, and high‐speed countercurrent chromatography were implemented for the accurate and targeted separation of active compounds. The inhibitory activities of target compounds to pancreatic lipase were detected by the inhibition experiments in vitro. The binding affinities and binding sites were analyzed using molecular docking.ResultsA total of eleven kinds of pancreatic lipase inhibitory substances were screened from C. tangutica. Seven triterpenoid saponins were screened for the first time as lipase inhibitors and successfully prepared with purities higher than 97%. Tanguticoside B, clematangoticoside J, hederoside H1, and rutin showed stronger inhibitory effects with IC50 values of 1.539 ± 0.048, 1.661 ± 0.092, 1.793 ± 0.069, and 1.792 ± 0.094 mmol/l. Moreover, they have the lowest ΔG values of −10.84, −9.97, −10.87, and −9.39 kcal/mol to pancreatic lipase.ConclusionThe integrated strategy using AU‐HPLC‐QTOFMS, targeted separation, in vitro validation, and molecular docking was feasible for rapidly screening and directionally isolating pancreatic lipase inhibitors from C. tangutica.