Abstract: This paper describes the determination of strychnine and brucine in the seeds, root, stem and leaves of Strychnos species by HPLC. The analytical column used was ZY110 YNG-C₁₈. The mobile phase was KH₂PO₄(0.01 mol·L^-1)—MeOH(73∶27), pH 2.5, regulated by 10% H₃PO₄. Flow rate was 1.0 ml·min^-1. The detection wavelength was 264 nm. The linear ranges of strychmine and brucine were 0.18～7.26 μg and 0.11～4.32 μg, respectively. The recoveries of strychnine and brucine were 98.27% and 98.04%, respectively. The analytical results showed that the contents of strychnine and brucine in samples showed great difference between different species. The contents of strychnine in the seeds of Strychnos wallichiana and S. ignatii were 5.6% and 3.9%, respectively. These results show that the two Strychnos species may be developed as the resources of strychnine.