Purpose: To investigate the antimicrobial efficacy of different chemical agents used for denture cleaning. Materials and Methods: Biofilm samples collected from 10 removable dentures were subjected to 10 disinfection protocols: distilled water for 30 min (negative control); 1% sodium hypochlorite for 10 min (positive control); diluted sodium hypochlorite for 10 min; vinegar for 20 min; 0.2% peracetic acid for 5 min; alkaline peroxide solution for 5 min; alkaline peroxide solution for 30 min; 0.12% chlorhexidine digluconate for 10 min; 0.05% sodium salicylate solution for 10 min; and enzymatic detergent for 2 min. Each of the samples was plated on petri dishes with Mueller-Hinton agar. The numbers of microbial colonies after 48 h at 37°C were compared using the Kruskal-Wallis and Dunn's tests (α = 0.05). Results: Diluted sodium hypochlorite, vinegar, and chlorhexidine digluconate inhibited bacterial growth, with an effect similar to that of 1% sodium hypochlorite. The 0.2% peracetic acid and 0.05% sodium salicylate solutions were ineffective against bacterial growth, while enzymatic detergent and alkaline peroxide achieved an intermediate effect. Conclusion: Diluted sodium hypochlorite, vinegar, and chlorhexidine digluconate can be considered adequate products for cleaning dentures due to their potential for inhibiting bacterial growth, similar to 1% sodium hypochlorite. However, the effect of these chemical agents on acrylic resin (polymethylmethacrylate) denture base materials needs to be examined to provide complete information about their clinical indication. Keywords: chemical cleaning, denture hygiene, microbiology, oral health