A novel fluorophotometric method for formaldehyde determination in environmental waters was developed: the method does not require any enrichment procedures. A flow injection analysis method for the spectrofluorometric determination of formaldehyde in waters, which is based on the reaction of formaldehyde with acetoacetanilide and ammonia, is proposed. The proposed method shows a good linearity from 0.50 to 40 x 10 -7 M, and the limit of detection (LOD) of 3 x 10 -9 M (0.09 ppb) is achievable. The sample throughput is 15 h -1 . One of the main advantages in the proposed method is that the reaction can be carried out at room temperature without any heating system. The effect of 1 various interferences possibly present in the real water samples was investigated. Most cations and anions, as well as organic compounds, do not interfere with the determination of formaldehyde in environmental water samples. The proposed method is very simple, rapid, less expensive, and highly sensitive, and can be applied to the environmental water samples, such as rain, tap water and river water, at low concentration levels without any enrichment procedure.