Objective： This study aimed to study the subchronic toxicity of acesulfame as a food additive and obtain the dosage of harmful effects not observed orally for 90 d. Methods： According to GB 15193.13—2015, eighty 4-week-old specific pathogen-free （SPF） healthy detached Sprague Dawley （SD） rats （half male and half female） were randomly divided into 4 groups. Rats were administered 4 500, 1 500, 500 and 0.0 mg/kg BW. acesulfame by gavage for 90 consecutive days, respectively. At the end of the experiment, samples of blood were collected for blood routine and biochemical analysis, and some organs were removed for histopathological examinations. All the data of the rats were compared and evaluated with the control group. Results： The general physical signs of animals in the 3 dosage groups of acesulfame 4 500, 1 500 and 500 mg/kg BW. were normal, and no significant abnormal changes were found in the body weight gain, total food intake, total food utilization rate, absolute organ weight, and viscera/body ratio at the end of the experiment. The values of hematology, biochemical indexes, and urine indexes were all within the normal fluctuation range, and there was no significant difference between the 3 experimental groups and the control group （P>0.05）. Gross examination showed no abnormal changes in the heart, liver, spleen, lung, kidney, testis, heart, thymus and adrenal glands. Histopathological examination also showed no pathological changes related to the subject. Conclusion： Acesulfame has no obvious toxic effects on rats during the whole test period, the NOAEL of male and female SD rats in oral subchronic toxicity study for acesulfame both are 4 500 mg/kg BW.