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 analyzed using Fisher's exact test.