A new steroidal alkaloid, hupehenizioiside (1), together with four known steroidal alkaloids hupehenizine (2), hupehenirine (3), peiminine (4) and hupeheninoside (5), were isolated and identified from the bulbs of Fritillaria lichuanensis. The structure of hupehenizioiside (1) was determined to be (20R,25S)-5alpha,14alpha,17beta-cevanine-6-oxo-3beta-O-beta-D-glucoside by spectral analysis and chemical evidence. Compounds (2)-(5) were isolated from Fritillaria lichuanensis for the first time.