OBJECTIVE: To isolate and identify the secondary metabolites of Cordyceps cardinalis for providing reference for the further exploration of the active substance of C. cardinalis and the natural product library of Cordyceps. METHODS: The compounds from C. cardinalis fermentation broth were isolated by silica gel column chromatography, C18 reverse phase silica gel chromatography, Sephadex LH-20 gel column chromatography, TLC and so on. The structures of isolated compound were identified by 1H-NMR, 13C-NMR and MS spectra. RESULTS: Six compounds were isolated from the ethyl acetate extract of C. cardinalis fermentation broth as 5-methyl-1, 3-benzenediol (compound 1), 4-hydroxy-17R-methylincisterol (compound 2), 5α, 8 α-epidioxy-(22E,24R)-ergosta-6,22-dien-3β-ol (compound 3),3β,5α,6β-trihydroxy-(22E,24R)-ergosta-7,22-dien (compound 4), uridine (compound 5), oosporein (compound 6). CONCLUSIONS: Six compounds are isolated from C. cardinalis strain, among which compound 1 and compound 2 are isolated from the genus Cordyceps for the first time.