We identified acaricidal compounds in Inula japonica Thunberg (Asteraceae) that were effective against carmine spider mite, Tetranychus cinnabarinus (Boisduval). Petroleum-ether extracts from I. japonica were toxic to T. cinnabarinus, with a 50% lethal concentration (LC₅₀) value of 1.18 mg/ml. Silica gel column chromatography was used to separate the acaricidal components. Seventeen of 38 fraction groups had mite mortality rates >50%, nine fraction groups had rates >60%, and three fraction groups had rates >80% at 2 mg/ml. The major volatile compounds in the bioactive fraction groups were identified by gas chromatography-mass spectroscopy, and β-sitosterol (1), stigmasterol (2), lupeol (3), and α-amyrin (4) were identified and determined to have acaricidal activity against T. cinnabarinus in vitro.