Abstract: OBJECTIVE To obtain the optimal conditions for separating the alkaloids from the extract of Hylomecon japonica by selecting appropriate cation exchange resins. METHODS Five types of cation exchange resins were evaluated in separating efficiency with measuring the adsorption ratio and eluting ratio of total alkaloids as indices, and the content of berberine hydrochloride from Hylomecon japonica was determined as an index by HPLC to choose the optimal technological parameters. RESULTS The optimal result of extraction was obtained as Hylomecon japonica shattered into thick powder, extracted with ten times 70% alcohol at room temperature for 2 hours and refluxing and extracting for 1 h (totally extracting for 3 times). C008 cation exchange resin had the good ability of absorbing the total alkaloid from Hylomecon japonica, and each gram of C008 cation exchange resin could absorb 0.65 g of the total alkaloids, and the desorption ratio was 13.81%. CONCLUSION The C008 cation exchange resin can be used for extracting total alkaloids from Hylomecon japonica and the established procedure is simple and feasible.