Objective: To establish a common method to detect the content of chitosan oligosaccharide. Methods: Chitosan oligosaccharide was hydrolyzed completely by concentrated hydrochloric acid, and the solution was regulated into neutral with NaOH. Then, determined the absorbance in 525nm, and substituted into the regression equation to determine the results. Results: The results showed there was a good linear relationship when the concentration of chitosan oligosaccharide ranged from 0.02 mg/mL to 0.12 mg/mL, r2 = 0.999. The average recovery of chitosan oligosaccharide samples was 99.25%. Conclusion: The method is sensitive, accurate and simple. It is applied to determine of the content of chitosan oligosaccharide.