Abstract: In environmental samples ⁹⁰Sr is one of the most important radionuclides generated in nuclear fission process. To improve the separation of ⁹⁰Sr from environmental water samples, the solvent containing crown ether was spreaded on the diatomite, the influence of parameters such as crown ether concentration, carrier mass, eluant volume etc on ⁹⁰Sr separation were studied based on former crown ether extraction method, and the crown ether extraction chromatography for ⁹⁰Sr separation was established. The experimental results show that the optimal crown ether concentration, ⁹⁰Sr carrier and eluant volume are 0.89mol/L,100mg and 200mL respectively. Through parameters optimization, the chemical recovery rate of Sr is over 90% when a new such chromatographic column is used. This ether extraction chromatography is satisfied with the conventional requirement for low-level ⁹⁰Sr separation and analysis in environmental water samples.