Objective To analyze the low-temperature disinfection effect of a compound peroxide disinfectant using glycerol and propanol as antifreeze agents. Methods Carrier quantitative germicidal test and object surface simulation on-site disinfection test were performed to analyze the disinfection effect and safety of the compound disinfectant at low temperatures. Results At constant temperatures of -20℃, -10℃ and 20℃, the logarithmic killing values of the compound peroxide disinfectant on Escherichia coli, Staphylococcus aureus and poliovirus on the carriers were >3. In cold storage at -20℃ and 10℃, logarithmic killing values of the compound peroxide disinfectant on both Staphylococcus aureus and Escherichia coli on the carriers at different sites were >3. The safety evaluation test showed that the acute oral toxicity lethal dose(LD₅₀)value of the compound disinfectant in mice was greater than 5 000 mg/kg. It did not cause irritation to rabbit skin, and had no damage on mouse chromosome. Conclusion The compound peroxide disinfectant with glycerol and propanol as antifreeze agents has a strong killing effect on bacterial propagules and viruses at low temperature, and is safe and non-toxic.