[Objective] To establish a convenient and accurate method for the content determination of polysaccharides from RHIZOMA GASTRODIAE. [Method] Extraction process of RHIZOMA GASTRODIAE polysaccharides was optimized by orthogonal test. The total saccharide content, uronic acid content and protein content were detected by phenol-sulfuric acid method, m-hydroxydiphenyl method, and Coomassie Brilliant blue method, respectively. [Result] The optimal extraction condition was A2B2C3, which were 70 ℃ extraction temperature, 2.5 h extraction time, and 3 extraction times. Under this condition, the extraction rate of polysaccharides reached 13.15%. Saccharide contents in different groups of RHIZOMA GASTRODIAE polysaccharides were 65.8%, 84.6% and 87.3%; protein contents were 2.9%, 0.28% and 2.7%; and uronic acid contents were 29.7%, 26.3% and 39.6%. [Conclusion] This method was rapid, simple, accurate, and sensitive, and could be used for the quality control of RHIZOMA GASTRODIAE polysaccharides. [Method] To establish a convenient and accurate method for the content determination of polysaccharides from RHIZOMA GASTRODIAE. [Method] Extraction process of RHIZOMA GASTRODIAE polysaccharides was optimized by orthogonal test. The total saccharide content, uronic acid content and protein content were detected by phenol -sulfuric acid method, m-hydroxydiphenyl method, and Coomassie Brilliant blue method, respectively. [Result] The optimal extraction condition was A2B2C3, which were 70 ° C extraction temperature, 2.5 h extraction time, and 3 extraction times. Extraction contents of polysaccharides reached 13.15% Saccharide contents in different groups of RHIZOMA GASTRODIAE polysaccharides were 65.8%, 84.6% and 87.3%; protein contents were 2.9%, 0.28% and 2.7%, respectively; and uronic acids contents were 29.7%, 26.3% and 39.6%. [Conclusion] This method was rapid, simple, accurate, and sensitive, and could be used for the quality control of RHIZOMA GASTRODIAE polysaccharides.