Objective: To study the effects of four separation and purification methods on total anthraquinone content in Radix et Rhizoma Rhei decoction. Methods: To Compare the purification efficiency of four methods: gelatim precipition, pH adjustment, polyamide adsorption and macropurous resin adsoption, solid content and total anthraquinone in Radix et Rhizoma Rhei decoction were selected as markers. Results: All solid content of Radix et Rhizoma Rhei decoction greatly decreased after purification while the total anthraquinone content was varied from one method to another. The total anthraquinone content retention rate of macroporous adsorption resin Ⅰ and Ⅱ were highest ( 93.21% , 95.63% respectively). Conclusion: Macropurous adsorption resin have a good enriching effect on the separation and purification of total anthraquinone in Radix et Rhizoma Rhei decoction.