Abstract: As a typical pollutant, N,N-dimethylformamide (DMF) is characterized by large discharge, high ecotoxicity, and poor degradability. However, there is still a lack of research on relevant criteria or standards for it at present. Based on relevant Chinese and English literature and toxicological databases such as ECOTOX, Web of Science, and China National Knowledge Infrastructure (CNKI), this study collected and screened 222 pieces of literature data on the effects of DMF for Chinese freshwater organisms. Meanwhile, the acute and chronic toxicity test data of DMF were also obtained for 8 native representative species, including Pseudorasbora parva, Pelteobagrus fulvidraco, Corbicula fluminea, Branchiura sowerbyi, Misgurnus anguillicaudatus, Brachionus calyciflorus, Brachionus angularis, and Chlamydomonas reinhardtii. These data were used for the derivation of DMF water quality criteria for freshwater organisms. The DMF water quality criteria involved a total of 26 freshwater aquatic species, covering three trophic levels. It also included economically important species widely distributed in China, such as Ctenopharyngodon idella and Aristichthys nobilis. Specifically, the acute toxicity data covered 24 species belonging to 15 families and 7 phyla, with the most sensitive species being Anabaena cylindrica and Anabaena variabilis; the chronic toxicity data covered 12 species belonging to 10 families and 6 phyla, with the most sensitive species being Ceriodaphnia dubia. The species sensitivity distribution (SSD) method and the toxicity percentage ranking (TPR) method were used to calculate the DMF water quality criteria for freshwater organisms in China. The short-term and the long-term water quality criteria derived by the SSD method were 824.12 and 14.53 mg/L; the short-term and the long-term water quality criteria derived by the TPR method were 1 081.58 and 22.48 mg/L, respectively. It was suggested that the criteria derived by the SSD method be adopted as the water quality standards for DMF.