Aim: The aim of the present study was to evaluate the effectiveness of three disinfectants as toothbrush decontaminants–4.5% vinegar,3% sodium hypochlorite, and 0.2% chlorhexidine gluconate. Material and methods: 60 participants in the age group of 20–30years were divided into 4 groups. Group I(4.5% vinegar), group II(3% sodium hypochlorite), group III (0.2%chlorhesxidine gluconate) and group IV(tap water), which served as the control group. Each participant was provided with a new toothbrush, instructions were given and toothbrushes were collected after brushing for seven days. Toothbrushes were sent to the laboratory for determining total colony count.Results:4.5% vinegar was most effective followed by 3% sodium hypochlorite and 0.2% chlorhexidine gluconate and tap water. Conclusion: We concluded that vinegar was most effective disinfectant when compared with others.