The chemical components and availability of Paris rugosa were investigated for the first time, using a UPLC-MS/MS-based molecular networking strategy and phytochemical research. Ultimately, eleven undescribed steroidal saponins, parisrugosides A-K, and ten known analogs were identified. Their structures were confirmed using comprehensive spectroscopic data and chemical methods. The aglycones of parisrugosides A-D are first spirostanes with an epoxy group at C-5/C-6, a hydroxy group at C-7, and a double bond at C-8/C-9 or C-8/C-14. Parisrugosides G and H possess an undescribed spirostane aglycone with two double bonds located at C-5/C-6 and C-8/C-9, which are conjugated with a carbonyl group at C-7. The isolates were evaluated for their cytotoxicity against five human cancer cell lines (human HL-60 leukemia, A549 lung, MCF-7 breast, SMMC-7721 liver, and SW480 colon solid cancer cell lines). Parisyunnanoside D, kingianoside K, and dichotomin displayed significant cytotoxicity against these cancer lines, with IC values ranging from 0.50 to 19.58 μM.
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