Recently, applications of high voltage impulse (hereafter HVI) technique to desalting, sludge solubilization and disinfection have gained great attention. However, information on how the operating condition of HVI changes the water qualities, particularly production of hydroxyl radical (·OH) is not sufficient yet. The aim of this study is to investigate the effect of operating conditions of the HVI on the generation of hydroxyl radical. Indirect quantification of hydroxyl radical using RNO which react with hydroxyl radical was used. The higher HVI voltage applied up to 15 kV, the more RNO decreased. However, 5 kV was not enough to produce hydroxyl radical, indicating there might be an critical voltage triggering hydroxyl radical generation. The concentration of RNO under the condition of high conductivity decreased more than those of the low conductivities. Moreover, the higher the air supplies to the HVI reactor, the greater RNO decreased. The conditions with high conductivity and/or air supply might encourage the corona discharge on the electrode surfaces, which can produce the hydroxyl radical more easily. The pH and conductivity of the sample water changed little during the course of HVI induction.