Potentilla anserina L. is not only a medicinal plant, but also a traditional cuisine. Hence, an acute toxicity study was performed to confirm its safety profile. Forty Kunming mice were randomly divided into two groups: control group and P. anserina L. extract group. Using the maximum dosage method, the P. anserina L. extract group was given the maximum dose within 12 h, equivalent to 345.6 g/kg crude drug. The control group was given distilled water. After administration, toxicity symptoms of mice were observed, body weight and food intake were recorded. After 14 days, blood was collected to measure biochemical parameters, autopsy was carried out to observe the changes of organs, and the vital organs were separated, weighed, and preserved for histopathological examination. The results showed that P. anserina L. extract group had no toxic symptoms. The activity, weight, and diet of mice were normal, and no abnormality was found in organ index, renal function, liver function, anatomical observation, and histopathological examination. Therefore, the maximum oral dosage (345.6 g/kg) of P. anserina L. was good safety. This study indicated that P. anserina L. had a large safety range and the clinical application was safe.