ABSTRACT Aim: The aim of the study was to evaluate if vinegar and vinegar with 3.5% sodium chloride could be used as an alternative to chlorhexidine gluconate for disinfection of toothbrushes. Materials and methods: The study consisted of three groups: group I: 0.12% chlorhexidine digluconate, group II: 38% white vinegar, and group III: 38% white vinegar with 3.5% sodium chloride. Two new toothbrushes were cultured to check their sterility before use. Eight children in the age group of 6 to 12 years were given oral hygiene instructions and four sets of oral hygiene kits. At the end of the 1st week, one set of used toothbrushes was cultured to check for total viable count. Again, at the end of the 2nd, 3rd, and 4th weeks, the 2nd, 3rd, and 4th sets of brushes were collected, subjected to respective decontamination treatment for 12 hours, and then cultured for microbial analyses. The obtained data were analysed using Fisher’s exact test. Results: In group I, three out of eight brushes showed 1 to 10,000 colonies, in group II, one out of eight cases showed >10,000 colonies and two out of eight showed 1 to 10,000 colonies, and in group III, all the eight cases showed no colonies upon culturing. Conclusion: Out of all the tested decontaminating agents, combination of 38% white vinegar and 3.5% sodium chloride was found to be the most efficient. Clinical significance: Toothbrushes become contaminated upon use and act as a reservoir for microorganisms. Thus, toothbrush decontamination should become a routine practice. Testing the disinfecting efficacy of vinegar and common salt is, thus, beneficial as they are available in every kitchen.