In order to explore the brassinosteroid-active components in Zea mays seeds, the methanol extract was purified by the sequences of solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 chromatography, charcoal adsorption chromatography and Bondesil chromatography. The activity of brassinosteroid was monitored by the rice inclination test and its presence could be confirmed in each purification step. The purified active components were separated by silica gel adsorption chromatography. Brassinosteroid substances in separated active fractions were identified as castasterone and teasterone by HPLC. The content of brassinosteroid in Zea mays seeds as converted into brassinolide was 3-8ng/g fresh weight.