The method for separation of tea saponin was investigated. A standard sample of tea saponin was obtained by silica gel column chromatography and the method of colorimetric analysis of tea saponin was then established. HPD series of macroporous resin were screened by static adsorption and elution of ethanol,and the content was high. The separation and purification of tea saponin with HPD400 macroporous resin by dynamic adsorption and ethanol gradient elution. The results indicated that the dynamic saturated adsorption capacity of HPD400 was 109.3 mg/g and the content of tea saponin of sample eluted by 30% ethanol and 50% ethanol were 93.1% and 87.1%, respectively. The total elution rate with ethanol was 80.3%. Tea saponin was eluted mainly by 30% ethanol,and the content was high. HPD400 macroporous resin is suitable for separation and purification of tea saponin.