Abstract: A new method for the determination of peimisine in bulbus Fritillariae by ion-pair HPLC has been developed. The chromatographic system consists of ODS column and mobile phase of methanol—water(69: 31) containing 7.5 mmol/L of SDS(pH 4.5±0.1). Twelve samples of different species in the family of Fritillaria have been analyzed by this method. The results show that all samples contain peimisine and the contents of peimisine are correlative to those of total alkaloids.