Toothbrushes get easily contaminated with different microorganisms originating not only from the oral cavity but also from the surroundings in which they are stored. Contaminated toothbrushes might serve as a possible cause in infection or reinfection especially in patients undergoing periodontal treatment. The purpose of the present study was to evaluate and compare the efficacy of five different disinfectant solutions like hydrogen peroxide (3%), Chlorhexidinegluconate (0.2%), essential oil, Saline (9%) and Cetylpyridinium chloride.Sixty dental graduates were randomly assigned as control and experimental groups and were provided with toothbrushes and the disinfectant solution for routine use twice daily for three days. They were instructed to immerse the toothbrush head in the disinfectant solution for five minutes after brushing and the toothbrush was air dried. Toothbrushes were collected from volunteers after three days of use for microbiological analysis. 3% Hydrogen peroxide (MD= -2.02, p<0.001), mouthwash containing 0.2% chlorhexidinegluconate (MD= -1.79, P<0.001) and mouthwash containing essential oils (MD=-1.51, P<0.008) gave a significantly lower CFUs score whereas saline and cetylpyridinium chloride mouthwash failed to produce a significant difference in the number of CFUs when compared with that of water. The presents study shows that using 3% H2O2 in 1:1 dilution for disinfecting toothbrush after brushing is the most effective method followed by mouthwashes containing .2% chlorhexidinegluconate and essential oils respectively.