Background:Toothbrushes are essential for eliminating dental biofilm and preventing caries and periodontal disease. Regular disinfecting is necessary to maintain a clean toothbrush. Chlorhexidine is the gold standard, but it may be resistant to periodontal pathogens. Hydrogen-rich water, due to its antioxidant properties, may be beneficial against periodontal infections.Method:A study comparing hydrogen water and chlorhexidine as toothbrush disinfectants was conducted. Thirty dental students were divided into two groups: A and B. Group A students used hydrogen water and Group B used chlorhexidine mouth rinse as a disinfecting solution. They were instructed to brush twice a day for seven days and disinfect their toothbrushes with the disinfection solution for 10 minutes. Bristles from the collected toothbrush were sectioned and incubated for 24 hrs and colony-forming units (CFU) were counted.Result:The results showed that hydrogen water had significantly less colony-forming units in brushes disinfected with hydrogen water compared to chlorhexidine.Conclusion:Hydrogen water has better antimicrobial properties than chlorhexidine mouthwash. Therefore, hydrogen water is recommended as a regular disinfectant for toothbrushes.