PROBLEM TO BE SOLVED: To obtain sterilization effect even under the condition of relatively low temperature while using an acid solution having relatively high pH.SOLUTION: A mixed liquor of at least either an acetate buffer solution or a lactate buffer solution with fumaric acid is used as a bactericidal agent. An bactericidal object is immersed into the one as the mixed liquor in which pH is regulated to the range of 3.5 to 5.5. Then, the temperature of the bactericidal object is controlled to the range of 40 to 70°C, and it is held for 1 to 15 hr.