To investigate the technological parameters of the isolation and purification of 10-hydroxycamptothecin and vincoside-lactam from Camptotheca acuminata seed by polyamide. The static arid dynamic adsorption characteristics of 10-hydroxycamptothecin and vincoside-lactam on polyamide were studied, and the contents were determined by HPLC. The optimum parameters for adsorption were as follows: the contents of 10-hydroxycamptothecin and vincoside-lactam in the extracts were 0.189 g x L(-1) and 0.334 g x L(-1), respectively, pH 6, flow rate was 1.0 mL x min(-1), processing volume was 3 BV; for desorption: ethanol-water (60:40), flow rate was 1.0 mL x min(-1), 5 BV as an eluent. After treated with polyamide, the contents of 10-hydroxycamptothecin and vincoside-lactam were 17.52% and 32.87%, respectively, the recovery yields were 66.05% and 75.86%, respectively. Results showed that polyamide revealed a good ability to separate 10-hydroxycamptothecin and vincoside-lactam. Therefore, we concluded that results in this study may provide scientific references for the large-scale production of 10-hydroxycamptothecin and vincoside-lactam extracted from C. acuminata seed.