Cell cultures of L929 and BHK21/C13 cells were used to evaluate the toxicity of a newly introduced bleaching agent (Colgate Platinum) compared to hydrogen peroxide, an established bleaching agent. The cell reaction was determined by a quantitative technique at 24 h and 72 h. Both bleaching materials had a dose-dependent effect on cell viability. Concentrations of hydrogen peroxide causing a 50% decrease in cell number (50% inhibition dose-ID50) were calculated as 0.00034% after 24 h and 0.00001% after 72 h in L929 cells. The ID50 of hydrogen peroxide was found to be 0.00016% after 24 h and 0.00007% after 72 h in BHK21/C13 cells. The ID50 of Colgate Platinum was 0.00074% after 24 h and 0.00045% after 72 h in L929 cells and 0.00055% after 24 h and 0.00024% after 72 h in BHK21/C13 cells. The results showed that, in vitro, both bleaching agents were cytotoxic to fibroblasts and the new bleaching agent was less toxic than hydrogen peroxide.