Recently, separation membranes have been applied to fields such as water supply, sewage treatment, gray water reuse, and air pollution control. Chemical cleaning technology is attracting attention among the methods of reusing these expensive separation membranes. It was found that the separation membrane could be regenerated using chemical cleaning. Specifically, it was found that the use time of the separation membranes regenerated by chemical cleaning was sustainable for more than 1,700 hours. Additionally, it was found that the flux recovery ratio after chemical cleaning was maintained at least 60%. In addition, the flux recovery ratio of HYDREX 4710, an organic membrane cleaner, and 4703, an inorganic membrane cleaner, was 76% and 62%, respectively, showing the highest flux recovery ratio among the chemicals used. Considering that the target raw water of this study is biological secondary treatment water, it was suggested that chemical cleaning could be actively used to regenerate separation membranes in future water treatment.