In this work, a simple and novel cloud point extraction (CPE) method for the separation and preconcentration of manganese in aquatic environment was developed and combined with flame atomic absorption spectrometry (FAAS). Manganese was formed a complex with Calcon carboxylic acid (CCA) in the presence of Cetyl pyridinium chloride (CPC) and Triton X-114. The main factors affected the CPE, such as pH, the dosage of Triton X-114, CCA, CPC, NaCl, coexisting ions, and the heating time and heating temperature. After optimization of the experimental conditions, the system of Mn-CCA-CPC was able to overcome the matrix interference and showed the significant effect of separation of enrichment. The precision of the method was 0.39% and the limit of detection was 0.27 μg·L-1 of Mn. The results of the determination of manganese content in tap water and groundwater were 33.5 and 64.5 μg·L-1, which was in line with the international standards, so the method can be successfully applied for the determination of manganese in the water environment.