Abstract: According to the nature of cyano hydrolysis and the basic principles of acid-base titration, a acid-base titration method is established for the determination of the content of drug intermediates 1, 6-Di(N3-cyano-N1-guanidino) hexane by an alkaline hydrogen peroxide procedure using a sulfuric acid standard solution to titrate the excess lye. The effects of heating time and the number of glass beads were investigated. The results show that the best experiment conditions are 1.5 hours of heating time and the using of two glass beads. The method is applied to determine four unknown samples with satisfactory results. The results are consistent with high performance liquid chromatography.