The oriental armyworm, Mythimna separata (Lepidoptera: Noctuidae) is a harmful pest which causes heavy loss in agricultural crops. Sex identification of M. separata is very important for understanding the biology and management of this pest. Development of fast and convenient methods to identify M. separata female and male is the necessary prerequisite for conducting gender-related studies. In this study, new methods to differentiate sex of M. separata at pupal and adult stages were investigated. Our results showed that posterior abdominal segments containing genital organs which are morphological traits to differentiate female and male. Genital openings are important characters for the female and male pupae differentiation. For female pupa, it locates on the ventral side of eighth and ninth abdominal segments and appears as a visible longitudinal suture; for male pupa, it locates on the ventral side of the ninth abdominal segment and appears as a short slit surrounded by a pair of visible semicircular lumps. Bristle numbers are another indicator for sex differentiation of adults. Each female has three-bristled frenulum and each male has one bristled-frenulum near the base of hind wing of each side of moth body. We confirmed the two sex differentiation methods manually by dissecting genitalia of adults and found their genger was as same as what we examined the sex of pupae and adults using the morphological methods.