Abstract: Objectives: To establish the optimal method of phenol-sulfuric acid method to determine polysaccharides from wind-steamed Polygonatum cyrtonema Hua. Methods: The anthrone-sulfuric acid method and phenol-sulfuric acid method were used at the same time to measure the polysaccharide content of wine-steamed Polygonatum cyrtonema Hua. For the phenol-sulfuric acid method, the concentration of phenol, the amount of phenol, the amount of sulfuric acid, the heating time and the heating temperature were selected for single-factor investigation. And the response surface method was used for analysis and optimization. Results: Analyzing the F value by response surface method, the influence of each factor on the absorbance was: The amount of sulfuric acid>the concentration of phenol>the amount of phenol. The heating temperature and time had no obvious effect. The best process obtained was 1.5 mL of phenol, 4.5% of phenol and 6.5 mL of sulfuric acid. Measuring by this condition, the average content of polysaccharides was 8.50% and the relative standard deviation was 0.87%. Conclusion: The phenol-sulfuric acid method after the optimization could accurately measure the polysaccharide content in the wine-steamed Polygonatum cyrtonema Hua, which would provide reference value for further study of quality standard.