Abstract: In order to determine polysaccharides of germination fungus HL- 003, parameters of the phenol-sulfuric acid method were optimized. Phenol concentration and volume of concentrated sulfuric acid was the independent variable, the maximum absorption wavelength and absorbance were detected under different concentrations of phenol and concentrated sulfuric acid volume. Immediately,the color of time,precision,linearity and recovery were measured in order to determine the optimum parameters. The results showed that phenol concentration of 5%,concentrated sulfuric acid content of 3.5 m L,the maximum absorption wavelength of 488 nm,coloration time 15 min. The methods had a high precision and recovery results were reliable,and consistent with the true value. The concentration of glucose in the range of 0~200 mg/m L had a good linearity.Intracellular and fermentation of polysaccharide contents were 2.028% and 2.998 g/L,by detecting with optimized method.