The separation and purification of steviosides by column chromatograph was studied. Adsorption capacity,desorption ratio,recovery ratio and purity were chosen as indexes,column separation method with alumina and many kinds of macroporous adsorption resin as adsorbent was performed. Static tests indicated that two kinds of macroporous adsorption resin,AB-8 and ADS-7,were suitable. However,dynamic testing showed the effect of ADS-7 was better than that of AB-8. The saturated adsorption capacity,desorption ratio and recovery ratio of ADS-7 were184. 29 mg / g,81. 66% and 74. 12%,respectively. ADS-7 was showed to be the best separation resin for preparing high purity stevioside as high as 96. 95% in the final product.